Search Results (1,353)

Infection of the large yellow croaker (Larimichthys crocea) embryo cell line YCE1 with megalocytivirus strain FD201807 leads to accumulation of capsid-deficient viral intermediates within intracellular vesicles at 48 h post-infection (a phenotype associated with non-lytic egress), which coincides with the initial peak of viral genomic copies. To characterize the host molecular response during this critical stage, we performed time-course RNA sequencing at 24, 48, 96, and 144 hpi. Integrated analysis identified 6661 differentially expressed genes (DEGs) and 1138 differential alternative splicing (DAS) events affecting 892 genes, with DAS event abundance peaking at 48 h. DAS genes in autophagy and Golgi vesicle transport pathways, both integral to animal innate immunity, were significantly enriched exclusively at this timepoint, featuring novel mutually exclusive exon (MXE) isoforms in gopc (Golgi-associated PDZ and coiled-coil motif containing) and rint1 (RAD50 interactor 1). Weighted gene co-expression network analysis (WGCNA) of DEGs identified mapk9 (mitogen-activated protein kinase 9) and map1lc3a (microtubule-associated protein 1 light chain 3 alpha) as hub genes within modules enriched for autophagy-related functions. Separate co-expression analysis of DAS genes revealed rnf5, rimoc1, and golga4 as hub genes, with gopc exhibiting only a single linkage to rnf5. These findings implied concurrent transcriptional and virus-induced host splicing regulation of vesicle-associated innate defense pathways and suggest that splicing-derived features may serve as potential candidates for diagnostics or prevention against megalocytivirus disease in L. crocea. Full article

Ischemic stroke induces complex molecular responses that disrupt subcellular organelles’ function and contribute to brain injury, yet the temporal changes of organelle-specific transcriptomic remodeling remain to be investigated. In this study, we performed in silico analysis of publicly available transcriptomic data from isolated brain microvessels of transient middle cerebral artery occlusion (tMCAO) mouse model. Using in silico approaches, we analyzed differential gene expression at 24 h (acute phase) and 7 d (intermediate phase) post-stroke, focusing on mitochondria, endoplasmic reticulum (ER), and Golgi apparatus. Functional enrichment (Gene Ontology, KEGG) and protein–protein interaction network analyses were performed. Our analysis of the data revealed that at 24 h post-stroke, all three organelles exhibited marked transcriptional remodeling, where mitochondrial pathways showed disrupted metabolic and redox regulation; ER pathways indicated activation of biosynthetic processes, stress signaling, and ferroptosis; and Golgi-related genes reflected altered vesicular trafficking and glycosylation. By 7 d, mitochondrial alterations subsided, whereas ER and Golgi pathways displayed downregulation of metabolic and neuronal signaling processes, indicating persistent dysfunction and incomplete microvascular recovery. Phase-specific drug–gene interaction analysis will be useful to understand temporal organelle-associated transcriptional organization and to guide future investigations of neurovascular remodeling after ischemic stroke. Full article

Cycloheptylprodigiosin is a promising anticancer candidate that induces cancer cell death accompanied by severe Golgi stress. Although the soybean oil-based optimized MB2216 medium produced a total prodiginine titer approximately three times that of the basal MB2216 medium, the overall production level remained limited. In addition, a substantial fraction of the pigments partitioned into floating oil droplets, hindering efficient recovery by simple centrifugation. In this study, a novel medium was rationally formulated based on genomic insights derived from homology analysis of conserved biosynthetic genes involved in cycloheptylprodigiosin production in Spartinivicinus ruber MCCC 1K03745^T. Sequential optimization through single-factor experiments, full factorial designs, steepest ascent experiments and response surface methodology identified an optimal medium consisting of peptone (5 g/L), yeast extract (1 g/L), peanut meal (7.611 g/L), and L-Proline (0.695 g/L) prepared in seawater at pH 7.6. Under the optimized conditions, the total prodiginine titer reached 53.33 mg/L, which was 11.37 times that of the basal MB2216 medium and 3.29 times that of the soybean oil-based MB2216 medium. Moreover, the pigment-associated biomass could be efficiently recovered by centrifugation. This study provides a genomics-informed strategy for improving prodiginine production in S. ruber and facilitates downstream pigment recovery. Full article

The cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway serves as a central innate immune signaling axis in host defense against DNA virus infections, and RNA viruses have also evolved diverse strategies to counteract this pathway. Encephalomyocarditis virus (EMCV), a zoonotic RNA virus, utilizes its 2C protein to antagonize RIG-I-like receptor-mediated type I interferon signaling and induce autophagic degradation of calcium binding and coiled-coil domain 2, thereby evading host antiviral immunity. However, the precise molecular mechanism by which EMCV 2C protein modulates the cGAS-STING pathway remains incompletely understood. Herein, we show that EMCV infection reduces the expression of cGAS and STING proteins, and its 2C protein significantly suppresses the production of IFN-β triggered by poly(dA:dT) or viral infection, as well as the mRNA expression of interferon-stimulated genes. Mechanistically, 2C protein binds to STING via its ATPase domain and facilitates K48-linked polyubiquitination and proteasomal degradation of STING, while dominantly interfering STING translocation to the Golgi apparatus and the formation of STING-TBK1-IRF3 complex, thereby blocking STING-mediated IFN-β signal transduction at multiple levels. This study reveals a novel mechanism by which the EMCV 2C protein suppresses the host antiviral response by targeting STING and promoting its ubiquitination and degradation. This finding deepens understanding of the immune evasion mechanism of EMCV and provides a theoretical foundation for the development of antiviral therapies targeting the 2C protein of picornaviruses. Full article

Background: A paediatric cancer diagnosis is a profound stressor for the entire family system. Although coping strategies are well-studied, their link to the quality of the parent–child attachment relationship remains less explored. In this study, we investigated whether dyadic attachment dynamics—specifically closeness and conflict between parent and child—are associated with the use of adaptive or maladaptive coping strategies in caregivers of children undergoing active treatment for oncohaematological diseases. Methods: We conducted a multicentre, cross-sectional study across three Italian paediatric oncohaematology centres. A total of 165 caregivers of 91 paediatric patients aged 3–17 years completed self-report measures assessing parent–child relationship quality (Child–Parent Relationship Scale-CPRS), coping strategies (COPE-NVI), perceived social support (MSPSS), and resilience (RS-14). We tested whether the quality of the parent–child attachment relationship is associated with caregivers’ coping strategies. We hypothesised that Attachment Closeness would be associated with adaptive coping (Positive Attitude, Social Support, Problem Orientation), whereas Attachment Conflict would be associated with maladaptive coping (Avoidance). We conducted multiple linear regression models, adjusted for key covariates and with robust standard errors clustered at the family level, to test these hypotheses. Results: Higher levels of emotional closeness (CPRS) were significantly associated with greater use of adaptive coping strategies, specifically Positive Attitude (β = 0.20, p = 0.049) and Problem Orientation (β = 0.26, p = 0.002), even after controlling for sociodemographic factors, social support, and resilience. Conversely, higher levels of relational conflict were significantly associated with greater use of the maladaptive Avoidance strategy (β = 0.14, p = 0.015). The hypothesis linking closeness to Social Support seeking was not supported. Conclusions: The findings suggest that the parent–child attachment relationship is a significant correlate of caregiver coping strategies in caregivers of children with cancer. Interventions aimed at supporting the caregiver–child dyad by fostering emotional closeness and reducing conflict may promote more adaptive parental coping mechanisms, thereby enhancing family resilience and psychological adjustment throughout the treatment journey. Full article

The increasing detection of micro-nanoplastics (MNPs) in environmental settings and in human biological samples has raised growing concern about their potential implications for human health. Exposure to plastic particles may cause oxidative stress, inflammation, and metabolic reactions, according to previous studies. It is unidentified, consequently, to what extent these basic processes result in identifiable clinical illness patterns. This narrative review investigated whether a structured symptom-mapping approach might be used to identify recurrent multisystem symptom patterns that might be consistent with environmental exposure. The Chicago Cluster System (CCS) is a conceptual framework that includes four environmental exposure indicators in addition to 26 clinical signs and symptoms. The CCS framework was used for filtering clinical descriptions of 375 disorders in 21 disease groups using conventional medical references. Using exploratory thresholds of at least eight compatible symptoms, the aim of this study was to identify scenarios exhibiting convergence with the CCS symptom pattern. Forty disorders (11%) exhibited eight or more CCS-compatible symptoms out of the 375 diseases that were evaluated. Neurodegenerative, metabolic, inflammatory, and gastrointestinal illnesses were among the various clinical categories in which these symptoms emerged. Fatigue (65%), upper gastrointestinal problems (58%), and increased inflammatory markers (55%) were the most commonly reported symptoms. These results imply that certain types of chronic disorders may exhibit recurrent multisystem symptom patterns. The observed overlap should be regarded cautiously because many CCS signs are non-specific and widely spread across medical conditions. The CCS framework may organize multisystem symptom patterns in environmental health research. Full article

Recently, a shift from substance-based to process-based ontologies of living beings and biological entities has been widely advocated, largely on the grounds that traditional substance thinking, by encouraging biological reductionism, fails to adequately capture the nature of biological wholes. Process-based approaches are instead taken to provide a more appropriate metaphysical framework for the constitutive dynamicity of living systems. These arguments, however, have been criticized for relying on overly reductive characterizations of substances, which both classical and contemporary accounts describe as inherently involving change and activity. In this essay, I address the substance-versus-process debate from the perspective of contemporary cell biology. I argue that conceiving the cell as a substance is not only compatible with the centrality of processes, but that the cell continues to function as the fundamental reference point in biology precisely because it entails processuality as intrinsic to its dynamic mode of being. Within this framework, subcellular entities are identified by their functional subservience to the cellular whole. On this basis, I propose an empirically grounded criterion for distinguishing between purely processual and substance-like subcellular entities. Processual entities, such as the Golgi complex and the nucleolus, lack dedicated repair systems and tend to disassemble upon inhibition of specific metabolic activities. By contrast, substance-like entities, including cell-derived organelles such as the mitochondrion and the nucleus, depend for their persistence on specific repair systems, and their eventual dismantling under non-permissive conditions cannot be straightforwardly understood as the mere interruption of a process, but instead appears as the outcome of an active, regulated response. Full article

The etiology of autism spectrum disorder (ASD) implicates genetic predispositions and environmental chemicals, such as polybrominated diphenyl ethers (PBDEs). We aimed to identify whether mitochondrial quality control (MQC) was involved in ASD-relevant behavioral changes induced by decabromodiphenyl ether (deca-BDE, BDE-209) and the alleviation by melatonin. Pregnant rats exposed to BDE-209 (50 mg/kg i.g.) were administrated melatonin through drinking water (0.2 mg/mL) during gestation and lactation. Behavioral assessments integrated open-field test, three-chamber social test, and Morris water maze; mitochondrial detections took transmission electron microscopy, immunofluorescence, and homeostasis together; hippocampal molecular network was identified through transcriptomics profiles, combining dendritic morphology analysis after Golgi-Cox staining. Melatonin supplementation attenuated BDE-209-reduced social and cognitive ability, accompanied by improvements in hippocampal synaptic plasticity (dendritic spines, PSD95, SNAP25). Mitochondrial dysfunctions, shown as decreases in complex IV activity, ATP content, and mtDNA copies, plus redox imbalance (ROS/SOD2) and resultant mitochondrial membrane potential disruption and apoptosis, together with fusion/fission dynamic (MFN2/DRP1), biogenesis (SIRT1-PGC1α-TFAM), and mitophagy (SIRT3-FOXO3-PINK1) suppression, were reversed by melatonin partially through SIRT1 (Sirtuin-1)-dependent pathways, as these protections were abolished by inhibitor EX527. This study highlighted the SIRT1–SIRT3 axis in MQC and behavioral effects, providing novel intervention for PBDEs’ neurodevelopmental impairment. Full article

Cancer remains a significant challenge to global public health. Preliminary studies indicate that the protein Golgi-associated, Gamma-adaptin Ear Containing, ARF Binding Protein 2 (GGA2) may influence various cancers. However, the potential role of GGA2 in oncogenesis remains unknown. We utilized data from The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) projects to analyze GGA2 expression levels. Genetic variations and protein expression of GGA2 in human tissues were assessed using the cBioPortal. Gene Set Enrichment Analysis (GSEA) provided deeper insights into GGA2’s oncogenic functions. Comprehensive analysis of TCGA datasets combined with ESTIMATE and TIMER tools demonstrated significant correlations between GGA2 expression levels and clinical outcomes, survival metrics, genomic instability markers (microsatellite instability (MSI)/tumor mutational burden (TMB)), and immune microenvironment composition. Functional validation in prostate cancer models employed qRT-PCR quantification, immunoblotting verification, and cellular behavior assessments through colony formation, Transwell migration, and wound closure assays. Our findings suggest GGA2 could serve as a prognostic biomarker in various cancers. Abnormal levels of GGA2 promoter methylation and genetic alterations may contribute to its dysregulated expression in some cancers. Distinctly, GGA2 expression correlates with MSI and TMB across different cancers and is linked to the expression of immune checkpoint genes. Functionally, GGA2 is instrumental in inhibiting oncogenic mechanisms by diminishing the proliferation, colony formation, invasion, and migratory capabilities of prostate cancer cells. Our study shows that the oncogenic role of GGA2 in various cancers and GGA2 could be served as a biomarker of PARD. Full article

The Golgi complex undergoes dynamic remodeling during the cell cycle, as ribbon unlinking in G2 is required for proper mitotic progression. Failure to fragment the ribbon leads to G2 arrest, whereas forced mitotic entry with intact ribbons results in multipolar spindle formation. Phosphorylation of the Golgi matrix protein GRASP65 at serine 277 (S277) in rat (S274 in human) by JNK2 is essential for ribbon unlinking, but its upstream regulation has remained unclear. Here, we generated and validated a phospho-specific antibody recognizing human GRASP65 phosphorylated at S274, enabling accurate detection of this modification. Using this tool, we identify protein kinase D2 (PKD2) as a critical upstream regulator required for GRASP65 phosphorylation and Golgi unlinking. GRASP65-S274 phosphorylation increases during G2 and is markedly reduced upon PKD2 inhibition or depletion, resulting in decreased Golgi unlinking and delayed G2/M transition. Conversely, PKD2-activating stimuli, including phorbol esters and nocodazole, enhance GRASP65 phosphorylation in a PKD2-dependent manner. These findings define PKD2 as a key regulator of the JNK2–GRASP65 signaling axis controlling Golgi disassembly at the G2/M transition. Moreover, the phospho-specific GRASP65 antibody described here provides a valuable tool to dissect upstream signaling mechanisms and to identify the initial triggers driving Golgi unlinking at G2 entry. Full article

The goal of this review is to bring to the attention of the scientific community the opportunities of transmission electron microscopy for analyses of biological subjects and resolving complicated cases of data interpretation. Although procedures for electron microscopy are in general more elaborate (particularly for simultaneous immunolabeling of multiple antigens) compared to fluorescent microscopy, they can help view cellular morpho-functional features undetectable using other methods. In this review, we consider several unexpected and serendipitous discoveries made in our laboratory and fulfilled using unique opportunities provided by electron microscopy of ultrathin sections. We are deliberating the following topics: interpretation of unusual results of immunolabeling; a novel method for in situ identification of cells undergoing mitochondrial disorder and necrosis-like death; the sequence of organelles’ reorganization in dying cells; simultaneous rupture of nuclear and plasma membranes in migrating neurons; and the role of cytoskeleton in lateral expansion of the cerebral cortex. Full article

Platelets are small circulating blood cells that mediate haemostasis and thrombosis. Platelets respond to vascular damage by adhesion, granule release, and aggregation. Healthy endothelial cells inhibit platelets through prostacyclin-induced cAMP signalling. Intracellular cAMP activates protein kinase A (PKA), a tetrameric kinase composed of two regulatory (R) and two catalytic (C) subunits. cAMP-binding triggers dissociation of C subunits from the PKA complex and phosphorylation of substrate proteins, which mediate platelet inhibition. The R subunits of PKA are known to be attached to A-kinase anchoring proteins (AKAPs), which enable subcellular compartmentalisation of cAMP signalling. Proteomics have identified 22 AKAPs in platelets, but only a few of these have been studied in detail. This review summarises current knowledge about platelet AKAPs, including studies done regarding other cells. Possible integration of AKAPs into platelet signalling is explored with a focus on subcellular localisation, interaction partners, and PKA-mediated substrate phosphorylation. As main platelet compartments, the plasma membrane, endosomes, mitochondria, the Golgi, the dense tubular system, and the cytoskeleton are considered. Potential roles of individual AKAPs in platelet inhibition are discussed, and open questions in the field are defined. Full article

Influenza A virus (IAV) remains a major global health threat, and host-directed antivirals may help overcome rapid viral mutation and drug resistance. Here, we performed a genome-wide siRNA screen in A549 cells using cell viability as an integrated endpoint to identify host determinants of IAV (PR8/H1N1) infection. Using plate-normalized viability ratios, we identified 2134 genes with >40% viability change after infection (2048 UP and 86 DOWN; two-tailed t-test, n = 3; p < 0.05, FDR < 0.1). MetaCore pathway analysis showed enrichment of programs linked to host response and tissue injury control, including RAS-related signaling and multiple metabolic pathways such as estradiol, ubiquinone/mitochondrial redox, and benzo[a]pyrene/xenobiotic metabolism. DAVID Gene Ontology analysis further highlighted biological processes relevant to infection, including endocytosis, transcription, and translation, consistent with host pathways supporting viral replication. Benchmarking against meta-analyzed RNAi and CRISPR resources revealed that shared hits were enriched for translation, nucleocytoplasmic transport, and ER-Golgi trafficking, supporting external validity, whereas the large unique UP fraction was dominated by hormone metabolism, detoxification, and mitochondrial redox/CoQ pathways, consistent with viability-specific, tolerance-associated host response programs. Integrating the screen with DrugBank identified 174 druggable host genes corresponding to 345 candidate compounds. Together, these findings provide a systematic resource of host factors influencing H1N1 infection, improve understanding of influenza virus–host interactions, and offer a foundation for future development of host-directed antiviral strategies and drug repurposing efforts. Full article

Background: Aging-related cognitive decline is closely associated with microglial senescence and the resulting chronic neuroinflammation. Emerging evidence identifies the cyclic GMP-AMP synthase–stimulator of interferon genes (cGAS-STING) pathway as a pivotal innate immune signaling pathway linking DNA damage to cellular senescence and the senescence-associated secretory phenotype (SASP), particularly in microglia. Targeting the formation or selective clearance of senescent cells thus emerges as a promising therapeutic approach to ameliorate cognitive dysfunction. Resveratrol has shown promise in modulating immune response and exerting anti-aging effects. However, the therapeutic potential and underlying mechanisms of resveratrol in mitigating age-associated microglial senescence and cognitive decline are not fully understood. Methods: In the present study, we employed a well-established murine model of accelerated aging induced by chronic intraperitoneal injection of D-galactose (D-gal) to elicit pronounced senescence-associated phenotypes and neuroinflammation. Resveratrol was administered via oral gavage daily for three weeks following D-gal injections. Behavioral assays were conducted to assess cognitive performance. Immunohistochemistry, quantitative PCR, and Western blot analyses were used to evaluate markers of cellular senescence, microglial activation and pro-inflammatory cytokine expression. In addition, in vitro assays in cultured microglia coupled with RNA sequencing were used to investigate the downstream signaling events following resveratrol treatment. Results: Chronic D-gal treatment induced significant cognitive impairment, enhanced microglial activation, elevated pro-inflammatory cytokine levels, and increased markers of cellular senescence in the brain. Resveratrol administration remarkably attenuated these effects, as evidenced by improved memory performance, reduced microglial senescence markers, and suppressed expression of Cxcl-10, Il-1β, and other SASP factors. Mechanistically, unbiased transcriptomic analysis revealed that the cGAS-STING signaling and neuroinflammation pathways were prominently dysregulated with double-stranded DNA-induced cellular senescence, which was effectively normalized by resveratrol in cultured microglia. Interestingly, resveratrol inhibited the translocation of STING from the endoplasmic reticulum to the Golgi apparatus and suppressed phosphorylation of TBK1, thereby blocking downstream STING signaling. Conclusions: These findings demonstrate that resveratrol mitigates microglial senescence and neuroinflammation and preserves cognitive function in D-gal-induced aging mice, at least partly through modulation of the cGAS-STING signaling. Therefore, targeting this pathway may represent a promising therapeutic strategy for age-related neuroinflammatory and cognitive disorders. Full article

Disruption of circadian rhythms caused by constant artificial lighting (“light pollution”) is a significant risk factor for the development of metabolic and age-associated pathologies. The liver, as a central metabolic organ with pronounced circadian regulation of its functions, is particularly vulnerable to desynchronosis. The aim of this study was to evaluate the effect of three-month dark deprivation (constant lighting) and the corrective action of exogenous melatonin on the morphofunctional state of the liver in young mature rats. The experiment used 3-month-old male Wistar rats, divided into groups: control (standard light:dark cycle 10:14 h), dark deprivation group (DD, constant lighting 24 h/day), and DD + Melatonin group (DD + Mel, dark deprivation with melatonin administered in drinking water at a dose of 12 mg/L). After 3 months (animal age 6 months), a comprehensive analysis was performed. It was shown that dark deprivation causes a profound (more than five-fold) suppression of plasma melatonin levels, which is accompanied by the formation of a pro-senescent and metabolically dysfunctional phenotype of the liver. This was manifested by the development of steatosis, an 18% increase in hepatocyte area, a 30% decrease in the proportion of binucleated hepatocytes, activation of cellular senescence markers (p16, p21) and stress markers (p53), and suppression of the expression of circadian transcription factors BMAL1 and CLOCK. At the ultrastructural level, lipofuscin accumulation, damage to mitochondria and the Golgi apparatus were noted. Biochemically, hyperglycemia, increased AST activity, hypoproteinemia, hypoalbuminemia, hypercholesterolemia, and hypertriglyceridemia were revealed. Administration of exogenous melatonin completely prevented the development of these disorders, normalizing hormone levels, morphology, ultrastructure, biochemical parameters, and the expression of key molecular markers. Thus, three-month dark deprivation induces complex pro-senescent remodeling and metabolic dysfunction of the liver, mediated by melatonin deficiency, while exogenous melatonin demonstrates a pronounced hepatoprotective and chronoprotective effect. Full article