Lactoferrin (Lf) is a multifunctional glycoprotein with antimicrobial, anti-inflammatory, and antioxidant properties that have been proposed as a therapeutic adjunct in wound healing.As keratinocytes are key drivers of re-epithelialization, this study investigated the effects of Lf on human HaCaT cells with a focus on viability, proliferation, migration, and oxidative stress. HaCaT cells were treated with Lf at concentrations of 5, 7.5, and 10 µg/mL, and outcomes were assessed using metabolic and proliferation assays, a scratch wound healing assay, and measurements of nitrite, thiols, and antioxidant enzyme activities. Lf preserved keratinocyte viability and did not induce uncontrolled proliferation, indicating good cellular tolerance. Migration was not affected at early time-points but showed modest enhancement at 48 hours in the 5 and 7.5 µg/mL groups. At the redox level, Lf selectively increased catalase activity across all doses, while nitrite and superoxide dismutase remained unchanged and thiols and glutathione S-transferase decreased at higher concentrations.Together, these findings identify Lf as a redox-centric modulator that enhances hydrogen peroxide detoxification without compromising cell integrity, supporting its potential as a safe adjunctive agent to promote orderly wound repair.

Osteoarthritis (OA) is a degenerative joint disorder characterized by chondrocyte dysfunction and progressive cartilage destruction, with ferroptosis emerging as a key contributor to disease pathogenesis. This study investigates the therapeutic potential of Aucubin, a natural iridoid glycoside, in OA treatment through modulation of the Keap1/NRF2 pathway and ferroptosis inhibition. Using an in vitro OA model established by IL-1β treatment of immortalized human chondrocytes (HCs), we demonstrate that Aucubin significantly improves cell viability, colony formation capacity, and migratory potential while promoting cell cycle progression. Aucubin also reduces ferroptosis-associated oxidative damage by decreasing MDA, GSSG, Fe2+, and ROS levels while increasing GSH content. Fer-1 ferroptosis inhibitor further supports this result. Molecularly, qPCR and Western blot analyses reveal that Aucubin upregulates NRF2 expression while suppressing Keap1 and modulating the levels of ferroptotic markers. These findings are further corroborated in a surgically induced OA rat model, where Aucubin administration attenuates cartilage degradation, restores Keap1/NRF2 expression, and normalizes ferroptosis-related protein expression. Our preliminary findings suggest that aucubin exerts its chondroprotective effects by inhibiting ferroptosis, at least partially through activation of the Keap1/NRF2 pathway, and may represent a potential therapeutic agent for osteoarthritis.

Chromatin remodeling plays a crucial role in gene expression. Chromatin architecture is governed by the interaction of a variety of proteins and transcription factors, including histones and non-histone chromatin-binding factors. Non-histone proteins, such as high mobility group-associated proteins (HMGA), are key players in this process. They do not have transcriptional activity per se but comprise flexible intrinsically disordered proteins (IDP) which interact with nucleosomes to change the compaction of chromatin at enhancers and promoters, thereby modulating the process of transcription. HMGA proteins have attained significant attention for their role in the regulation of gene expression during development, cell differentiation and in cellular senescence. Their molecular interactions are controlled by posttranslational modifications which determine nucleoprotein complex formation and function. This review highlights the role of HMGA proteins in nuclear organization, at telomeres and centromere regions and in senescence-associated heterochromatin foci (SAHF) and links these spatiotemporal chromatin architectural functions to the molecular domain structure of HMGA proteins in fine-tuning dynamic chromatin states.

Rosacea is a chronic inflammatory skin condition marked by excessive M1 macrophage polarization and angiogenesis, resulting in erythema and tissue inflammation. Despite available treatments, many patients experience recurrent flare-ups. This study explores chelerythrine, a bioactive component of Phellodendri Chinensis Cortex, for its therapeutic potential in rosacea through modulation of NF-κB, p38 MAPK and STAT3 signaling, inflammation, and vascular regulation.

Pseudogenes, traditionally considered non-functional gene copies, have garnered attention due to emerging evidence of their transcription and translation. Ubiquitin is canonically expressed from UBA52 and RPS27A genes as fusion proteins, with additional polyubiquitin precursors encoded by UBB and UBC. Several pseudogenes of these loci are annotated as non-functional. Here, we report that the RPS27A pseudogene, RPS27AP5, expresses two proteins: a ubiquitin variant (UbP5) and a ribosomal protein variant (S27aP5). These proteins mature through cleavage and exhibit localization and biochemical characteristics similar to their parental counterparts. S27aP5 integrates into ribosomes, and its overexpression leads to an increased 80S monosome fraction. Using affinity purification and polysome profiling, we show that S27aP5-containing ribosomes exhibit altered mRNA associations. The findings suggest that RPS27A, a processed pseudogene, can give rise to a ribosomal protein variant capable of integrating into monosomes and influencing mRNA association aligns with growing evidence that ribosomes may exhibit functional diversity.

Research is undergoing a paradigm shift. High-impact discoveries frequently require multidisciplinary approaches and are increasing in technological complexity. The days in which a single trainee can master all of the knowledge and technical skills necessary to complete a project by themselves are passing. In this evolving landscape, scientific cores-centralized facilities that provide advanced technologies and expert guidance-are becoming indispensable to the research pipeline. In this editorial, we suggest how core facilities in academic centers can evolve to meet these changes in research expectations by acting as full-service facilities, or like academic contract research organizations. In this new model, full-service cores will offer comprehensive project support, including the execution of the experiment. This paradigm shift will speed discovery, but requires modifications to existing research culture, including changing lab and project management approaches, increased recognition of the role of core directors and revised training expectations. Investigators and trainees will be expected to master narrow analytical but broad conceptual domains, while scientific cores will provide the technical and multidisciplinary expertise required to generate complex datasets. Revising the existing model will also require significant financial investment from host institutions and funding agencies. While initially challenging to implement, we predict that early adopters of this new model will be at the forefront of scientific discovery.

Recently, proteomics analyses using databases of unannotated ORFs revealed that ubiquitin (Ub) variants can be encoded and expressed from pseudogenes. One such pseudogene, UBBP4, produces Ub, which contains four substitutions (Q2K, K33E, Q49K, and N60S) relative to canonical Ub. Unlike Ub, Ub does not promote proteasomal degradation through K48 linkages and instead modifies a distinct set of proteins. To elucidate the structural basis of this divergence, we solved the NMR solution structure of Ub and characterized its backbone dynamics by 15N-relaxation. While Ub retains the overall helix-grip fold, we observed significant rearrangements and amplified motions in residues governing the Ub pincer mode, a conformational switch that determines whether UIMs engage the canonical I44 interface or the α1-β3 edge. Specifically, Q2K and K33E cooperate to enhance motions on both fast (ps-ns) and slow (µs-ms) timescales within α1, the β1-β2 loop, and β5-regions central to pincer mode regulation. In addition, Q49K, adjacent to I44, perturbs UIM recognition and likely interferes with K48 chain formation and binding to the proteasomal receptor S5a. Collectively, our findings identify structural and dynamical determinants that explain Ub distinct substrate profile and inability to target proteins for degradation.

Increasing evidence has indicated that transforming growth factor beta 1 (TGFB1) is engaged in tumorigenesis and progression. Nevertheless, the underlying role and mechanism of TGFB1 in stomach adenocarcinoma (STAD) chemotherapy remains unknown. TGFB1 levels in various types of cancers were first analyzed by the TCGA database. Next, the degree of cellular damage, apoptosis and autophagy were detected by lactate dehydrogenase kit, flow cytometry, autophagy fluorescence analysis, and Western blot assay. The gene highly correlated with TGFB1 expression was searched by LinkedOmics and KEGG. We disclosed TGFB1 was enhanced in STAD. Besides, TGFB1 was remarkably higher in STAD patients in oxaliplatin (OXA) chemoresistant group than sensitive group. Additionally, the half maximal inhibitory concentration (IC50) values of OXA-resistant cells were markedly elevated. Furthermore, TGFB1 reduced AGS-OXA and HGC27-OXA cell injury, inhibited apoptosis and induced cellular autophagy. The addition of the autophagy inhibitor 3-methyladenine hindered this phenomenon. Further studies revealed that muscle RAS oncogene homolog (MRAS) is a downstream target gene of TGFB1. TGFB1 accelerated MRAS level in OXA cells, and MRAS knockdown reversed the effects of TGFB1 on OXA cell function. TGFB1 induces cellular autophagy via MRAS, thereby promoting STAD OXA resistance.

This study investigates the protective effects of polysaccharides (LBPs) on X-ray radiation-induced damage in rat spinal cord neurons (SCNs) and examines whether this protection is mediated through the activation of autophagy. In vitro and in vivo experiments revealed that high-dose radiation significantly reduced SCN viability and colony-forming ability. However, treatment with 40 mg/L LBP markedly increased cell survival and autophagy levels. Immunohistochemistry and Western blot (WB) analyses demonstrated a significant upregulation of autophagy-related proteins, protein 1 light chain 3-II/I and Beclin-1, in the LBP intervention group. In vivo studies further showed that LBP reduced oxidative stress markers, such as malondialdehyde, and enhanced superoxide dismutase activity in spinal cord tissue. These findings indicate that LBP mitigates neuronal damage caused by ionizing radiation via autophagy activation and antioxidative mechanisms, highlighting its potential as a radioprotective agent.

Immune checkpoint inhibitors (ICIs)-related myocarditis, a severe complication characterized by elevated cardiac troponin I, poses significant clinical challenges in distinguishing it from acute myocardial infarction (AMI). Our study aimed to identify plasma protein biomarkers that differentiate ICIs-myocarditis from AMI. Plasma samples from 5 ICIs-myocarditis patients (with paired baseline and diagnosis samples) and 5 angiography-confirmed AMI patients, matched for age, gender, smoking history, and pre-existing heart disease, were analyzed using label-free liquid chromatography-mass spectrometry proteomics. A total of 1521 plasma proteins were identified, with 1325 quantifiable. Proteomic profiling revealed differentially expressed proteins (DEPs) in ICIs-myocarditis associated with myocardial contraction, proteasome activity, NF-κB signaling, immunoregulation, and amino acid metabolism. Through validation in animal models of ICIs-myocarditis and AMI, two plasma proteins-MYOM3 (myomesin 3) and galectin-1 (LGALS1)-were identified as potential biomarkers linked to the onset of ICIs-related myocarditis. Further validation using expanded clinical cohorts confirmed their differential expression. These findings highlight MYOM3 and galectin-1 as promising biomarkers for distinguishing ICIs-related myocarditis from AMI, providing insights for clinical diagnosis and mechanistic research into immune-related cardiotoxicity.

The XVIth International Conference on Lactoferrin, held in Rome in November 2023, showcased cutting-edge research on the multifunctional glycoprotein lactoferrin (Lf). Known for its broad antimicrobial, anti-inflammatory, and immunomodulatory properties, Lf continues to challenge the one-protein-one-function paradigm. Presentations highlighted its evolving therapeutic applications, including anti-biofilm strategies, modulation of immune responses, iron homeostasis, gut health, and cancer inhibition. Studies emphasized the importance of Lf source, iron saturation, and formulation, including recombinant and peptide derivatives. The collection underscores the expanding biomedical relevance of Lf and sets the stage for continued exploration at the upcoming XVIIth Conference in Mazatlán, Mexico.

Determination of IC values at a fixed substrate concentration ([S]) is frequently used to rank the potency of enzyme inhibitors and estimate inhibitor concentrations ([I]) to use in full inhibition analyses, particularly for structure-activity studies wherein the mode of inhibition is often known. Assays at an [S] yielding the greatest difference between the initial rates observed in the absence () and in the presence () of an inhibitor (i.e., - ) will increase the sensitivity for the detection of enzyme inhibition. For noncompetitive and uncompetitive inhibitors of single-substrate enzymes, - increases with increasing [S]; however, for competitive and linear mixed-type (LMT) inhibitors, - obtains a maximum at a specific "optimal" substrate concentration ([S]). Equations are derived describing the dependence of [S] on [I], the dissociation constant for the inhibitor (), and the Michaelis constant for the substrate (). For example, for competitive inhibition, [S] =  . For [I]/ values typically employed for inhibition studies (e.g., 0.5 ≤ [I]/ ≤ 4), [S] ≈ 2 or 3 will generally maximize the - difference for competitive or LMT (α ≥ 7) inhibitors, respectively. For competitive inhibition of bireactant enzymes, the "optimal" substrate concentrations depend on the Michaelis constants for both substrates, [I]/, and the concentration of the second substrate.

In eukaryotic cells, phospholipid asymmetry is actively maintained, with phosphatidylserine (PS) typically confined to the inner leaflet of the plasma membrane (PM), due to the active performance of the PS flippase ATP11/CDC50A complex. However, in the tumor microenvironment (TME), PS exposure on the outer leaflet occurs from multiple sources, including apoptotic tumor cells, necrotic tissue, viable endothelial cells, and tumor-derived exosomes. Especially, in apoptotic cells, the PS scamblase Xkr8 mediates PS externalization. This exposed PS plays a crucial role in immune suppression within the TME. PS binds to receptors on phagocytes, primarily macrophages and dendritic cells (DCs), triggering efferocytosis (the engulfment of PS-positive cells, usually apoptotic cells) and promoting anti-inflammatory responses.

Ferroptosis, a recently discovered form of cell death, plays an important role in cancer progression. It has been reported that ferroptosis plays a complex regulatory role in the biological processes of glioma. In glioma, the long noncoding RNA, HOXD cluster antisense RNA 1 (), functions as an oncogene, contributing to glioma progression. However, its potential functions in ferroptosis are unclear. Herein, we aimed to clarify the biological function and detailed molecular mechanism of in regulating ferroptosis in glioma. Firstly, cell viability, reactive oxygen species (ROS), and malondialdehyde (MDA) content assays were detected. The mechanisms of 's effect on ferroptosis were evaluated by detecting glutathione, Cys, and solute carrier family 7 member 11 (SLC7A11) levels. RNA immunoprecipitation and RNA pull-down techniques were employed to explore whether can directly bind AMP-activated protein kinase (AMPK). Our findings indicated that levels were augmented significantly in glioma tissue. knockdown induced MDA and ROS accumulation, subsequently resulting in ferroptosis. Further molecular analysis showed that the binding between and AMPK regulated the mechanistic target of rapamycin kinase pathway, inhibit the transport of Cys, and decrease the production of glutathione, eventually resulting in ferroptosis. Our study revealed that regulates cell ferroptosis, thus its downregulation might be an effective strategy to suppress glioma.

Ovarian cancer is one of the most common and lethal malignancy tumors in women. Chemoresistance is one of the main reasons for ovarian cancer relapsing. Understanding the regulatory mechanisms of chemoresistance generation is critical to develop novel therapeutic strategies. Here, we found that TRIM46 was upregulated in ovarian cancer cells and tissues with chemoresistance and associated with poor outcomes. Functional assays showed that TRIM46 promoted cisplatin (CDDP) chemoresistance. Furthermore, TRIM46 interacts with PI3K/AKT pathway inactivator pleckstrin homology domain leucine-rich repeat protein phosphatase 2 (PHLPP2) and downregulated PHLPP2 level. Treating with PI3K/AKT pathway inhibitor significantly reversed the effects of TRIM46-overexpressing on chemoresistance. In summary, our study reveals that TRIM46 promoted chemoresistance via downregulating PHLPP2, leading to activating PI3K/AKT pathway. This study provides a novel potential target for ovarian cancer therapy.

Galanthamine, an alkaloid derived from the Amaryllidaceae family, serves as an acetylcholinesterase inhibitor. Due to its central cholinergic properties, this compound is being actively studied as a potential treatment for Alzheimer's disease. However, the broader scope of its biological effects remains poorly understood. In this study, we explored the therapeutic potential of galanthamine in promoting neuronal differentiation and enhancing neurite outgrowth in neural stem and progenitor cells (NSPCs). Our detailed analysis demonstrated notable changes in neuronal morphology and complexity during maturation following galanthamine exposure. Notably, the compound significantly increased the proportion of neurons with multiple neurites, indicating its ability to stimulate neurite formation and foster the development of complex neuronal networks. Furthermore, galanthamine treatment led to a marked rise in the number of mature-appearing neurons, distinguished by elongated and intricate dendrites, highlighting its potential to enhance neural plasticity and repair mechanisms. Importantly, we also identified that galanthamine facilitates neuronal differentiation in NSPCs by up-regulating the insulin-like growth factor 2 signaling pathway. Collectively, these findings provide valuable insights into galanthamine's role in Alzheimer's disease and emphasize its promise as a therapeutic agent for this neurodegenerative disorder.

Cellular aging, driven by oxidative stress, mitochondrial dysfunction, and inflammation, is exacerbated by a high-glucose and high-lipid (HGHL) diet, leading to collagen degradation and skin aging. Psilocybin, a naturally occurring compound, has shown potential in reducing symptoms of aging. This study explores the protective effects of psilocybin on BJ-5ta fibroblasts exposed to HGHL, focusing on cellular viability, apoptosis, senescence, the inflammatory responses, and wound healing. First, fibroblasts were exposed to 25 mmol/L glucose and 400 µmol/L palmitic acid to establish cell aging. Then, psilocybin effects were tested in co- and post-treatment with HGHL. Post-treatment with psilocybin at 15 µmol/L (P15) and co-treatment with psilocybin at 10 µmol/L (P10) preserved cellular viability and decreased beta-galactosidase activity. P10 was most effective in reducing apoptosis and alleviating HGHL-induced S phase arrest. P15 also reduced senescence markers and decreased the expression of inflammatory cytokines , , and . Additionally, psilocybin promoted nonsignificant fibroblast migration, and P10 co-treated with HGHL significantly upregulated gene expression. These findings suggest that psilocybin's antioxidative, anti-inflammatory, and regenerative properties make it a promising natural compound for reducing skin aging, particularly under oxidative stress conditions. Further research is needed to explore its long-term effects, optimal dosages, and clinical applications.

Colorectal cancer (CRC) is a prevalent and malignant tumor of the digestive system, characterized by high incidence and mortality rates. This study aimed to investigate the heterogeneity of the tumor microenvironment (TME) and the involvement of immune cells in CRC. Single-cell RNA sequencing (scRNA-seq) data obtained from the Gene Expression Omnibus database were used to analyze and identify six major cell types across normal, core, and border tumor samples. A total of 27 414 cells from various regions of patients with CRC were selected for subsequent analyses. Cellular interaction analysis revealed that differential signaling pathways between the TME and normal tissues, with several pathways involving interactions between myeloid cells and epithelial cells. Myeloid cells were extracted and classified into six subtypes based on markers identified in the literature. Monocle3 revealed the trajectory of tumor-associated macrophages (TAMs) and identified genes associated with pseudotime. Single-Cell ENrichment analysis for Interpreting Cellular Heterogeneity analysis identified specific regulons and target genes associated with TAMs. This study reanalyzed single-cell RNA-sequencing data and provided insights into the heterogeneity of the TME, particularly in relation to the role of TAMs.

Once considered a metabolic "dead end", lactate is now increasingly recognized as a key mediator of diverse biological functions. Elevated lactate concentrations are associated with many physiological stress and strain responses, suggesting a role in homeostatic control. Recent evidence also indicates that lactate may suppress intrinsic metabolic heat production in mammalian cells. However, the mechanisms underlying this effect-and its implications for cellular responses to heat stress-remain unclear. Here, we (1) extend the observation of lactate-mediated heat suppression to baker's yeast (); (2) link this phenomenon to increased mitochondrial Complex II (succinate dehydrogenase) activity; and (3) demonstrate a corresponding reduction in heat-induced cell mortality. These results support an evolving view of lactate, offering new insight into its role in metabolic control and thermoprotection.

Filamins are dimeric actin-binding protein that play a critical role in mechanical signaling. They contain a mechanosensory region (MSR) that naturally folds into a globular closed conformation. Under mechanical stress, the MSR unfolds into an open conformation, exposing binding sites for numerous proteins. Filamins are involved in diverse cellular functions, and their mechanical binding targets are highly context-dependent. In this study, we employed AlphaFold2 modelling for screening proteins that specifically recognize the open conformation of filamins. We focused on the filamin, Cheerio, and conducted a biased screen to identify mechanical binding proteins. We selected the top 132 hits from the initial screening for further characterization. All identified binding proteins specifically recognize the open conformation of the MSR and not the closed conformation. Interestingly, the binding regions of these proteins lack obvious sequence similarity. While some false positives were identified, they could be effectively filtered out based on the secondary structure formed at the binding interface. This study provides a framework for identifying specifically filamin interactions in mechanosignaling.

Astronauts experience the reactivation of latent viruses in spaceflight, an indicator of reduced immunity. It is unclear how the immune system responds to pathogens in a microgravity environment. A longitudinal profile of leukocytes' transcriptome changes from participants to an Earth model of microgravity and from astronauts sojourning aboard the International Space Station revealed a reduced expression of immune-related genes while in microgravity. In the current study, we identified transcriptomic changes specific to the transition to and from bed/space, as well as the adaptation, and the recovery from microgravity/space exposure. The expression of immune-related gene shifted in opposite direction at phase transition compared to within the bed rest and reambulation phases. Differential expression of cytokine genes supported a reduced immune-response during the head down tilt bed rest phase and return to baseline levels at reambulation. Immunoglobulin gene expression increased after participants left the facility. The enrichment analysis of the differentially expressed genes identified the gene ontology terms virus/viral and genes previously involved in the modulation of the response to latent reactivation, including IFNL1, TNFSF14, IL10, and ISG15. Leukocytes' transcriptomic analysis revealed dynamic changes of immune-related gene expression timed with phases of spaceflight. The current analysis combined with previous evidence of herpesvirus reactivation during space mission represent a valuable model for the study of viral latency in vivo.