Search Results (1,349)

Background: Extracellular vesicles (EVs) mediate intercellular communication in the central nervous system and are a major source of biomarkers. This study characterizes the EV-derived proteome secreted by human endothelial brain cells (HEBCs), astrocytes, and neurons to identify cell-specific roles in intercellular communication in the brain. Methods: Mass spectrometry analyses of EVs and corresponding parent cells were performed to identify differentially enriched proteins. Gene Ontology (GO) analysis of statistically significant, abundantly expressed proteins between EVs and parent cells (log₂ fold-change ≥ 2.0, p < 0.05) was performed to assess cell-specific functions. Results: Proteome analysis identified on average 932 proteins in astrocyte EVs (versus 1725 in parent cells), 1040 in HEBC EVs (versus 5451 in parent cells), and 470 in neuronal EVs (versus 578 in parent cells). The analysis indicated that astrocytes had the highest number of significantly abundant proteins (118), followed by HEBCs (24) and neurons (25). Astrocyte EVs were enriched in lipoproteins, complement factors, and protease inhibitors; HEBCs EVs in tight junction proteins, adhesion molecules, and protease regulators; and neuronal EVs in chromatin-associated histones, tubulin isoforms, and RNA-binding proteins. Conclusions: The proteomic signatures of EVs from different neurovascular unit cells suggest specialized roles in blood–brain barrier homeostasis, immune regulation, and synaptic and epigenetic signaling under healthy conditions. These baseline signatures provide a framework for future studies to investigate how brain cell-derived EVs may contribute to neurodegenerative disorders. Full article

Crop losses driven by fungal pathogens remain a major constraint to global food production, reinforcing agriculture’s dependence on fungicide-based disease control. Soil acts as a long-term reservoir and key hotspot for the evolution and persistence of antifungal-resistant Fusarium. The intensive, prolonged use of overlapping single-site fungicides in agriculture strongly selects for both intrinsic and acquired resistance in soilborne Fusarium populations, contributing to major crop losses, food insecurity, and One Health concerns. This review synthesizes current knowledge on (i) target-site (CYP51, β-tubulin, cytochrome b, SDH, myosin-5) and non-target-site (ABC/MFS efflux, multidrug resistance, epigenetic regulation) resistance mechanisms across the genus Fusarium; (ii) the influence of management practices and fungicide characteristics and behaviour in soil in reshaping microbial communities and selecting for resistant Fusarium; (iii) the consequences for plant disease management and the limitations of practices like cultural and biological control; and (iv) innovative strategies for plant disease management, as well as the monitoring and detection of antifungal resistance in soils. These aspects show that soil reservoirs of antifungal-resistant Fusarium are compromising fungicide-based control and increasing risks across sectors, highlighting the urgent need for sustainable, multi-layered, integrated pest management strategies combined with robust, molecularly informed resistance monitoring. Full article

Food deprivation induces intestinal adaptations in Triatoma pallidipennis, a hematophagous insect with intermittent feeding habits. The ability to survive long periods without food promotes the persistence of this vector in the environment and contributes to its evolutionary success. This study employed one- and two-dimensional electrophoretic techniques combined with Western blot to evaluate the abundance of α-tubulin, β-actin, and the heat shock protein HSP70. These proteins were more abundant in the anterior midgut tissue of unfed insects than in that of fed insects. As these responses were similar in females and males, the observed adaptations primarily depend on feeding status and intestinal region. These findings provide further insight into the intestinal physiology of T. pallidipennis, a vector of the flagellate parasite Trypanosoma cruzi, the causative agent of Chagas disease. Full article

Plant architecture is a critical agronomic trait in watermelon (Citrullus lanatus), with vine length directly influencing planting density, light interception, and field management efficiency. Short-vine forms have become important agronomic targets in breeding due to their advantages of high-density planting, efficient light utilization, and simplified field management. In this study, a dwarf mutant, designated PKH207, was identified from an ethyl methanesulfonate (EMS)-mutagenized population of the watermelon inbred line G42. The mutant exhibited significantly reduced plant height and shortened internodes due to decreased cell expansion in stem tissues. Genetic analysis indicated that the dwarf phenotype in PKH207 is controlled by a single recessive gene, which was named Cldw2 (Citrullus lanatus dwarf mutant 2). Using a population of 558 F₂ plants, bulked segregant analysis sequencing (BSA-seq) and linkage mapping delimited the causal locus to a 540.6 kb region on chromosome 10. Within this interval, a single-nucleotide polymorphism (SNP) mutation was identified in the gene ClG42_10g0100600, encoding an α-tubulin, and this gene was determined to be the candidate gene for the dwarf phenotype. Transcriptome analysis revealed that this mutation significantly disrupts key biological processes, including cell wall biosynthesis, microtubule cytoskeleton organization, and auxin signaling pathways, contributing to the dwarfism phenotype. This study identifies a novel dwarfing allele in cucurbits and provides a direct molecular target for breeding compact watermelon cultivars suited to high-density production. Full article

Background: Colchicine is a microtubule-targeting anti-inflammatory agent with emerging relevance in cardiovascular disease; however, its effects on injury-induced vascular remodeling remain incompletely defined. Methods: In this study, a rat iliac artery clamp injury model was used to evaluate the effects of colchicine (0.5 mg/kg/day, oral gavage) over 28 days. Histomorphometric, histopathological, and immunohistochemical analyses were performed to assess vascular remodeling. In parallel, molecular docking and STRING/Cytoscape-based protein–protein interaction (PPI) network analyses were conducted to provide structural and systems-level context. Results: Colchicine significantly reduced intimal thickness, the intima-to-media (I/M) ratio, luminal stenosis, adventitial thickness, and collagen deposition, while preserving the lumen area and improving the remodeling index. Medial thickness was not significantly affected. Proliferative activity showed a decreasing trend without statistical significance. Circulating inflammatory cytokines, including TNF-α and IL-1β, did not differ significantly between groups. Docking analyses suggested potential interactions with β-tubulin, ADAM17, NLRP3, IKKβ, and RELA, while network analysis identified an interaction architecture centered on NF-κB-related regulatory components and inflammasome-associated signaling pathways. Conclusions: Colchicine attenuates injury-induced vascular remodeling in this experimental model. These findings, together with complementary in silico analyses, suggest a multi-target, inflammation-associated framework involving NF-κB-related and inflammasome-linked pathways. The in silico analyses provide supportive mechanistic context but do not establish causal relationships. Full article

With global cancer cases projected to reach 35 million by 2050 and drug resistance to existing chemotherapeutic drugs remaining a significant threat in cancer therapy, accounting for up to 90% of chemotherapy failures, the search for novel anticancer compounds continues to be increasingly important. This systematic review (2021–2025) examined the role of thiazoles and 4-thiazolidinones/thiazolidinediones as popular scaffolds in existing anticancer drug design. While researchers continue to focus on well-established molecular targets, such as EGFR, VEGFR-2, and tubulin, there is a notable difference regarding other preferred choices for thiazoles and 4-thiazolidinones/thiazolidinediones. Among analyzed mechanisms of anticancer activity notably favored for thiazoles was the inhibition of serine/threonine protein kinases (CDK-2, BRAF^V600E), while for 4-thiazolidinones/thiazolidinediones more studied were ROS generation and PPARγ activation. Furthermore, less-researched mechanisms of anticancer activity with no FDA-approved drugs such as PTP1B, SIRT2, PKM2, eIF4E, CA XI and XII inhibition for thiazole derivatives and pan-PIM kinase and BAG3 protein inhibition for 4-thiazolidinones/thiazolidinediones were evaluated as well. Notable was the popularity of the multi-targeting approach for modern drug design, with ~30% reporting two or more targets for their compounds. Despite these advancements, the review identified critical gaps in ADMET evaluations, safety analyzing against normal human cells and the lack of mechanistic studies connecting the targeted protein and the compounds anticancer effects. Full article

Toxoplasma gondii, the causative agent of toxoplasmosis, a disease widely distributed, is an intracellular parasite that invades host cells of different tissues using specialized endocytic activity. Recent studies suggest that tunneling nanotubes (TNTs), thin cell-surface projections, may participate in the parasite–host cell interaction. Here we report results that suggest the involvement of host-cell TNTs in the adhesion of T. gondii tachyzoites to epithelial LLC-MK² cells. Microscopy analysis showed that incubating cells in a medium containing 0.45 M sucrose induces reversible assembly of TNTs without affecting cell viability. The presence of extended TNTs correlated with increased parasite adhesion and reduced parasite entry, thus suggesting a structural or signaling role in mediating adhesion. TNTs assembled following sucrose incubation contain both actin and tubulin components as determined by immunofluorescence microscopy. These results highlight a possible functional relevance of TNTs in T. gondii host cell interaction, especially in parasite adhesion, opening new perspectives for understanding T. gondii-host cell interaction. Full article

Assessing genotoxicity, specifically gene mutations and chromosomal aberrations, is fundamental to chemical risk assessment. Notably, the early identification of an aneugenic mechanism is of crucial importance, allowing, in principle, for a threshold-based risk assessment approach. To investigate this issue while pushing towards innovation in risk assessment by leveraging New Approach Methodologies, in silico approaches stand out as a particularly promising avenue. Building on these premises and given the lack of QSAR models for aneuploidy in the public domain, the present study exploited the genotoxicity-relevant alert lists implemented in the OECD QSAR Toolbox to base the investigation of structure-activity relationships for aneuploidy. To address the lack of relevant structured data resources, a dataset of 65 confirmed aneugenic substances was specifically curated and designed for the study. The results highlighted widely differing performances among the various profilers, confirming a general limited discriminatory power for aneuploidy. On the other hand, a granular analysis of the results from individual structural alerts enabled the successful isolation of some features associated with the aneugenic mode of action. Moreover, a subset of tubulin-binding chemicals was investigated to determine whether targeting a specific protein improves the characterization of toxicological alerts. The findings provide a refined definition of specific toxicity determinants for tubulin binders and serve as a promising tool for early hazard assessment, potentially informing relevant AOPs. While the computational approach appears promising, the overarching challenge that emerges is the limited availability of well-curated experimental data. In fact, reliable data on aneuploidy are scarce and fragmented across the literature. Furthermore, existing compilations of micronucleus study results are often complicated by conflicting interpretations. Full article

Euonymus bungeanus is a highly valued ornamental tree/shrub species widely utilized in landscaping and afforestation in Northeast Asia, yet molecular studies on this species remain limited due to the lack of validated reference genes for reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). In this study, 16 candidate reference genes were selected based on classical plant reference genes and our previous transcriptome data. Their expression stability was comprehensively evaluated using 64 samples collected from diverse tissues and plants subjected to various abiotic stress/hormone treatments across multiple time points. Across all samples analyzed, PBG1 (20S proteasome beta subunit G1) exhibited the highest overall expression stability, followed by VAPD (vacuolar ATP synthase subunit D) and EIF4A (eukaryotic translation initiation factor 4A). For tissue-specific analysis, TSR2 (pre-rRNA-processing protein), VAPD, and PBG1 demonstrated the greatest stability. Under specific stress conditions, PBG1 and EIF4A were identified as the most stable genes under low- and high-temperature conditions. PP2A (protein phosphatase 2A) and TUB6 (beta-6 tubulin) were optimal for drought stress, while TSR2, SRP (nuclear speckle splicing regulatory-like protein), and PBG1 exhibited superior stability under salt stress. These findings establish a validated panel of reference genes enabling accurate and reliable gene expression normalization in E. bungeanus, thereby facilitating future functional genomics studies in this economically and ecologically important species. Full article

Endometriosis is characterized by enhanced cellular proliferation, migration, and resistance to apoptosis, contributing to lesion persistence and progression. Targeting cellular plasticity and mesenchymal-associated functions may therefore represent a promising therapeutic strategy. Here, we investigated the effects of Pelargonium graveolens essential oil (PGEO) on proliferation, apoptosis, migration, cytoskeletal organization, transcriptional regulation, and metabolic alterations in human endometriotic 12Z cells. PGEO treatment suppressed proliferative capacity in a concentration-dependent manner and significantly impaired cell migration, accompanied by reduced β-tubulin expression and decreased levels of mesenchymal-associated markers CD73 and CD105. Increased GRP78 expression together with ultrastructural alterations, including cytoplasmic vacuolization and mitochondrial and endoplasmic reticulum changes, indicated activation of cellular stress responses. Although transcriptional analysis revealed increased CCND1 and PIK3CA mRNA levels, these changes did not parallel the observed suppression of proliferation, suggesting compensatory regulatory responses. Untargeted metabolomic profiling revealed alterations in energy metabolism characterized by increased levels of glycolysis-related metabolites, reduced levels of several amino acids including glutamine and histidine, and changes in lipid-associated metabolites. Collectively, these findings demonstrate that PGEO suppresses proliferative and migratory behavior in endometriotic cells while modulating cytoskeletal, transcriptional, and metabolic pathways, highlighting its potential as a candidate for further investigation in endometriosis-targeted therapeutic strategies. Full article

Parkinson’s disease (PD) is a common neurodegenerative disorder marked by progressive loss of dopaminergic neurons in the substantia nigra pars compacta and the accumulation of Lewy bodies, intracellular inclusions enriched in α-synuclein. Synphilin-1 interacts with α-synuclein, localizes to Lewy bodies, and has been implicated in inclusion formation and neuroprotection in cellular and animal models; however, its physiological function in vivo remains poorly defined. Here, we generated and characterized a synphilin-1 knockout (Sph-1 KO) mouse by targeted genetic deletion of the Sph-1 locus and performed a comprehensive phenotyping battery including behavioral testing as well as biochemical, histological, structural, and ultrastructural analyses. Sph-1 KO mice survived to nearly two years of age and showed normal body weight, lifespan, motor performance, learning and memory, anxiety-like behavior, attention, and gross brain morphology. Western blot analyses indicated that levels of α-synuclein and synaptic proteins were largely unchanged. While outer mitochondrial membrane proteins were unaffected, the mitochondrial matrix protein HSP60 was reduced, consistent with altered mitochondrial proteostasis in the absence of synphilin-1. Strikingly, histochemical analyses, magnetic resonance imaging, and electron microscopy revealed early-onset hydrocephalus in Sph-1 KO mice associated with severe loss and disorganization of motile ependymal cilia in the ventricular lining, a cell type that normally expresses high levels of synphilin-1. Ultrastructural and immunohistochemical analyses revealed disrupted ependymal architecture, mislocalization of acetylated α-tubulin to the cytoplasm, cellular swelling, and enlarged, aberrant mitochondria, whereas cortical neurons appeared largely structurally unaffected. Together, these findings identify synphilin-1 as a key regulator of microtubule organization and cytoskeletal/organelle homeostasis in ependymal cells, required to maintain motile ciliogenesis, cerebrospinal fluid flow, and ventricular integrity. This unexpected role for synphilin-1 in ciliated brain epithelia, along with a reduction in the critical mitochondrial chaperone HSP60, broadens our understanding of synphilin-1 biology and provides a new framework for its potential relevance to PD-associated pathology. Full article

Pyrazole derivatives have emerged as an important class of heterocyclic compounds in anticancer research due to their structural versatility and broad spectrum of biological activities. This review provides a concise overview of recent advances in the development of pyrazole-based anticancer agents, with emphasis on synthetic strategies, structure–activity relationships, and molecular mechanisms of action. Common synthetic approaches, particularly condensation and cyclization reactions, have enabled the preparation of structurally diverse pyrazole derivatives for biological evaluation. Available evidence indicates that the type and position of substituents within the pyrazole scaffold markedly influence anticancer potency, selectivity, and target affinity. Reported compounds act through multiple mechanisms, including inhibition of cancer-related targets such as tubulin, epidermal growth factor receptor (EGFR), cyclin-dependent kinases (CDKs), Bruton tyrosine kinase (BTK), and deoxyribonucleic acid (DNA)-associated pathways, as well as induction of apoptosis and disruption of cell-cycle progression. Several pyrazole derivatives have shown promising activity in in vitro and in vivo models. Overall, the findings summarized in this review identify the pyrazole scaffold as a valuable platform for the design and optimization of novel anticancer agents and support its continued exploration in medicinal chemistry. Full article

Objectives: This study aimed to investigated the role of Giardia extracellular vesicles (EVs) in intercellular communication and to evaluated their potential as vaccine candidates. Methods: The immunomodulatory effects of Giardia EVs were assessed in mouse macrophages and human monocyte-derived dendritic cells (Mo-DCs), with a particular focus on key inflammatory signaling pathways. In vivo immunogenicity was evaluated following EV administration, and the antigenic composition of EV cargo was characterized by proteomic analysis. Results: Giardia EVs activated pro-inflammatory signaling pathways in mouse macrphages, including SAPK/JNK, ERK1/2, and NF-κB. This activation was associated with IκB-α degradation and nuclear translocation of p65. Furthermore, EV stimulation significantly upregulated the expression of pro-inflammatory genes, including Il1β, Il6, Il4, Ptgs2, Nos2, and Tnf, with log₂ fold changes ranging from 3.9 to 15.8. Consistently, EVs increased iNOS protein expression (28–45%) and nitrite production (9.6–12.3-fold). In human Mo-DCs, Giardia EVs promoted cellular maturation, as evidenced by increased expression of MHC-II, CD80, and CD86, and enhanced T-cell proliferation with a Th1-skewed profile. In vivo immunization induced antigen-specific antibody responses, with IgG subclass distribution indicative of a balanced Th1/Th2 response. Proteomic analysis identified immunoreactive EV-associated proteins, including elongation factor 1-alpha, α-7.3 giardin, tubulin, and variant surface proteins (VSPs), which are well-established antigens in Giardia infection, with prominent bands observed at approximately 22 kDa and 50 kDa. Conclusions: Collectively, these findings demonstrate that Giardia EVs modulate innate immune responses in vitro, elicit antigen-specific humoral immunity in vivo, and contain conserved immunogenic proteins. These properties support their potential as a promising cell-free vaccine platform against giardiasis. Full article

Mitotic catastrophe refers to a complicated mechanism of cell death characterized by failure to complete the processes of mitosis correctly due to aberrant chromosome segregation and abnormal tubulin polymerization. Post-translational modifications (PTMs) play a crucial role in the functional diversity of the proteome by mediating the covalent attachment of functional groups to proteins, which regulates the proteolytic cleavage of subunits, facilitating the degradation of entire proteins. Recent studies suggest that PTMs of key proteins are closely implicated in the occurrence, regulation and potential therapeutic targets of mitotic catastrophe. Here, we summarize how multiple PTMs, including phosphorylation, ubiquitination, acetylation, methylation and other types of PTMs, regulate mitotic catastrophe. In addition, potential therapeutic approaches targeting mitotic catastrophe were also discussed. It is anticipated that the inducement of mitotic catastrophe can serve as a promising new therapeutic approach for various diseases in the future. Full article

Background/Objectives: Osteoarthritis has been increasingly described as associated with systemic inflammation, raising the question of how it would affect fertility in young women with or without reproductive hormone administration. We studied oogenesis in mice with collagenase-induced osteoarthritis (CIOA) as a model system with fewer ethical limitations after estradiol (E2) or follicle-stimulating hormone (FSH) treatment. Methods: Oocytes have been isolated from mice subjected to various treatment regimens. The meiotic spindle, the chromatin, and the actin cap were fluorescently labeled and analyzed. Results: In addition to reduced maturation rates, specific oocyte abnormalities were registered when CIOA, FSH, or E2 were applied in isolation. Combined treatments showed that the spindle, chromatin, and actin cytoskeleton parameters were differently affected in oocytes from groups with CIOA treated by estradiol and those treated with FSH. Enlarged spindles, ooplasmic tubulin asters, aligned metaphases, and predominantly normal actin caps, often with an actin halo, were typical for groups with CIOA combined with estradiol. The groups with CIOA and FSH had slightly enlarged spindles, unaligned metaphases with degenerated chromatin surrounded by a cloud of depolymerized tubulin, and small actin caps. Conclusions: Our results show that experimental osteoarthritis with or without exogenous reproductive hormones negatively affects oogenesis, presumably due to systemic inflammatory factors making the ovarian microenvironment less capable of supporting oocyte maturation. Estradiol supplementation does not benefit oogenesis. FSH treatment induced cytoskeletal and chromatin abnormalities that presumably disturb the fertilization and development potential of affected oocytes. These data can have implications for assisted reproduction in cases of patients with osteoarthritis. Full article