Eosinophilic esophagitis (EoE) is a chronic immune-mediated disorder of the esophagus. However, the T-cell transcriptional programs distinguishing active disease from remission remain incompletely understood. In this study, we applied single-cell RNA sequencing to esophageal biopsies obtained from normal subjects and patients in remission or with active EoE, to characterize T-cell heterogeneity, state-dependent transcriptional changes, and intercellular communication networks. After stringent quality control, no substantial batch effects were observed. We identified five major T-cell subsets, with natural killer T (NKT) cells representing the most abundant population. Notably, naïve T cells and T helper 2 (Th2) cells were detected exclusively in active EoE samples, whereas regulatory T (Treg) and Th17 cells were present in all groups. The abundance of Tregs surpassed that of Th17 cells in active EoE but was similar to Th17 levels in normal and remission tissues. Functional enrichment analysis revealed a preferential association of NKT cells with receptor-binding functions. Th2 and naïve T cells shared signatures related to ribosome biology; naïve T cells additionally exhibited 5'-UTR binding and translation-regulator activity, aligning with their maturation potential. Consistent with the non-malignant nature of EoE, copy-number variation signals were minimal. Cell-type-specific differential expression analysis uncovered activated immune programs in NKT cells from active EoE, enriched for cytokine activity, along with epigenetic and transcriptional alterations in Tregs, including demethylase-linked functions, and protein-folding-related changes in Th17 cells. Cell-cell communication inference indicated a substantial rewiring of interaction networks in active EoE, suggesting activation of IL-16, IL-10, TRAIL, and PECAM1 pathways and heightened outgoing signaling from Th2 cells. Across all conditions, NKT cells served as dominant signaling senders and receivers. Collectively, these results delineate the T-cell composition and signaling circuits specific to active EoE, offering mechanistic insights into the maintenance of disease activity.
Background microRNAs play critical roles in modulating the molecular pathways involved in liver fibrosis. miR-107 has been reported to be elevated in the liver of patients with non-alcoholic fatty liver disease (NAFLD), and it is believed to regulate the PTEN gene, a negative regulator of the PI3K/PTEN/Akt signaling cascade. The PI3K/Akt pathway responds to metabolic stimuli like insulin and growth factors, influencing key functions such as lipid and glucose metabolism. In addition, hypoxia-inducible factors (HIFs), particularly HIF-2α, are also involved in liver metabolism and may contribute to lipid synthesis by activating the same signaling axis. The present study aimed to examine the gene expression patterns of miR-107, PTEN, Akt, and HIF-2α in liver tissue of patients with simple steatosis and cirrhosis. Methods This case-control study assessed the gene expression levels using quantitative real-time PCR (qRT-PCR) in liver tissues from individuals with simple steatosis (n = 6), cirrhosis (n = 32), and histologically normal controls (n = 7). Results miR-107 expression was significantly increased in cirrhotic tissues compared with controls (p < 0.05). The increase was mostly related to viral cirrhosis. PTEN expression was reduced in both disease groups; however, this decline reached statistical significance only in the NASH cirrhosis (p < 0.01). Also, HIF-2α and Akt gene expression decrease and increase in cirrhosis, respectively. No correlation was detected between miR-107 and PTEN gene expressions in liver tissues. Conclusion These findings suggest that there are complex interactions between miR-107 and PTEN gene expression. Apparently, Increased miR-107 expression in viral cirrhosis and reduced PTEN expression in NASH cirrhosis suggest potential associations with specific cirrhosis etiologies.
Long-term exposure to ambient particulate matter ≤2.5 μm in aerodynamic diameter (PM2.5) has been associated with optic neuropathy and retinal neurodegeneration, but experimental evidence remains limited. We assessed two certified environmental particulate reference materials in primary mouse retinal cells and after repeated ocular-surface exposure in mice. For the in vitro analyses, technical replicate wells were first averaged within each independent cell isolation, and each experimental condition was then normalized to the corresponding control from the same biological replicate. Particulate exposure produced condition-dependent changes in 2',7'-dichlorofluorescin diacetate (DCFDA) fluorescence and ATP-dependent viability, with statistically significant effects concentrated in selected retinal glial-cell conditions. After within-experiment normalization and multiplicity adjustment, direct comparisons did not demonstrate a statistically significant protective effect of α-tocopherol or N-acetylcysteine on particulate-associated responses, and retinal glial-cell coculture did not significantly improve RGC survival relative to monoculture. In postnatal mice, topical particulate administration was associated with inner retinal thinning and lower RGC-marker-positive cell counts. In adult Thy1-CFP mice, neither sample-specific nor post hoc pooled within-mouse comparisons with contralateral saline-treated eyes were statistically significant. A separate post hoc exploratory between-animal comparison showed fewer CFP-positive cells in pooled particulate-treated eyes than in independent bilateral-saline controls (mean difference, -14.4 cells; 95% CI, -28.1 to -0.7; P = 0.040). These findings support cellular and retinal susceptibility under the conditions used, but they do not establish particle transport to the retina, a causal oxidative mechanism, or quantitative risk from ambient PM2.5 exposure in humans.
Objective:To clarify the core regulatory role and molecular mechanism of farnesoid X receptor (FXR) in the progression from liver inflammation/fibrosis to hepatocellular carcinoma (HCC), and to provide a theoretical basis for targeted prevention and treatment of liver diseases. Methods:A systematic review was performed to integrate FXR's key regulatory mechanisms in bile acid homeostasis, inflammatory signaling, metabolic reprogramming, tumor pathways, and the tumor immune microenvironment. Intervention effects of natural compounds and FXR agonists/antagonists were analyzed for clinical translational potential. Results:FXR acts as a core regulator throughout the inflammation-fibrosis-HCC axis via bidirectional mechanisms. (1) In inflammation/fibrosis, FXR activation upregulates BSEP, FGF15/19, and SHP, inhibits CYP7A1 to maintain bile acid homeostasis, suppresses the TLR4/NF-κB axis and NLRP3 inflammasome, reduces IL-6/TNF-α, and directly inhibits hepatic stellate cell activation and matrix deposition. Agonists like INT-767 block progression from chronic hepatitis to fibrosis/cirrhosis (2) In HCC, FXR dysfunction (e.g., HBx C40-mediated). causes bile acid accumulation, glucose metabolic disorders, and Notch1/STAT3 activation, driving HCC progression. FXR activation inhibits STAT3 phosphorylation, glycolytic enzymes, and Wnt activity. Quercetin and obeticholic acid suppress tumor growth via FXR signaling. (3) Aberrant FXR signaling correlates with immune microenvironment dysregulation and immune escape. FXR agonists reshape the microenvironment and enhance anti-PD-1 efficacy; combined FXR and GPC3 detection improves HCC diagnostic specificity. Conclusion:FXR is a central hub regulating the liver inflammation-fibrosis-HCC axis. FXR-targeting agonists, natural compounds, and combination immunotherapy hold significant translational potential. Further studies are needed to clarify FXR's tissue-specific functions and bidirectional mechanisms, and to develop highly selective modulators for precise liver disease prevention and treatment.
metaKEGG is a comprehensive software package designed to streamline the visualization and integration of pathway enrichment results from multi-omics data, providing accessible and detailed insights into the molecular mechanisms driving health and disease. Unlike standard pipeline approaches, metaKEGG incorporates novel concepts allowing for clear, granular representation of gene-level or transcript-level expression changes. Beyond transcriptomic analysis, metaKEGG also supports epigenetic and regulatory metadata layers, such as methylation profiles and miRNA target annotations, offering users a versatile solution to depict complex regulatory interactions within a single pathway map. Its modular architecture provides nine analysis pipelines to suit various experimental designs, from comparing gene expression across multiple conditions to the integration of compound-based metabolomics data. Its implementation in Python ensures easy adoption and reproducibility, while a user-friendly web app allows researchers with limited bioinformatics expertise to harness metaKEGG's full potential.
Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, underscoring the need to better understand tumor-intrinsic mechanisms underlying aggressive disease progression. Aberrant glycosylation is increasingly recognized as a key regulator of cancer cell behavior; however, the functional contribution of α2,8-sialylation to CRC remains poorly defined. Here, we investigated the role of the α2,8-sialyltransferase (ST8SIA6) in CRC progression and its impact on mucin-mediated signaling. Analysis of human CRC tissue arrays showed that ST8SIA6 expression was reduced in higher-stage tumors. ST8SIA6 silencing enhanced migration and invasion and was associated with EMT-like molecular changes. Using Siglec-7-based lectin enrichment and proteomic analysis, we identified MUC16 as a prominent Siglec-7-reactive glycoprotein associated with ST8SIA6. Loss of ST8SIA6 reduced Siglec-7 reactivity of MUC16 and increased its cell-surface localization. Increased membrane-associated MUC16 was accompanied by enhanced association with integrin β4 and increased Src and FAK phosphorylation. Blockade of integrin β4 attenuated Src/FAK activation and suppressed CRC cell migration, supporting a functional role for this axis. In vivo, ST8SIA6-deficient CRC cells exhibited increased lung tumor colonization following tail-vein injection in xenograft models, accompanied by elevated membrane MUC16 and increased MUC16-integrin β4 colocalization. Consistent with these findings, human CRC specimens from advanced stages exhibited increased MUC16 expression and integrin β4 colocalization compared with early-stage tumors. Together, these findings support a model in which loss of ST8SIA6 enhances MUC16-integrin β4 signaling and contributes to aggressive CRC phenotypes through increased MUC16 membrane localization and integrin β4-dependent signaling. These findings identify a previously unrecognized glycosylation-associated signaling axis that warrants further investigation as a biomarker and potential therapeutic target in colorectal cancer.
Introduction Fertility depends on the complex process of folliculogenesis, yet many aspects of early human follicle development remain unclear. Immunohistofluorescence (IHF) enables detailed analysis of follicular structures and allows quantification. However, it is often affected by tissue autofluorescence. This study compares four commercially available kits to identify the most effective method for reducing autofluorescence and improving IHF accuracy. Methods Sections of human ovarian tissue were treated according to the staining protocol without antibodies, but with one of four kits reducing autofluorescence to compare their efficacy. Images were taken on the Ti2-E fluorescence microscope in three different fluorescence channels.The kit with the highest reduction in autofluorescence was applied for a strong and a rather subtle staining in order to analyze the signal-to-background ratio (SBR). Results All tested reagents reduced tissue autofluorescence in all three fluorescence channels. Among the evaluated products, TrueBlack showed the greatest reduction in autofluorescence. When combined with IHF staining, it significantly reduced background fluorescence and improved the SBR. Nevertheless, TrueVIEW and Ready Probes were similarly effective. Conclusion Autofluorescence in human ovarian tissue can be effectively reduced using commercially available reagents. Among the tested products, TrueBlack showed the highest efficacy with our specific protocol and setup. Given the high demand for research and the rarity of suitable human ovarian samples, sharing these results may support more accurate and reproducible immunofluorescence analyses in future studies.
The Notch1 signaling pathway, via the Notch1 receptor and its endogenous ligands, has been suggested to play a role in heart development, as mutations in Notch1 and associated genes are associated with abnormal development of the myocardium, aortic valve, and outflow tract. The cell lineages receiving this signaling are, however, still unclear, partly owing to technical difficulties in labeling Notch1 signaling rather than Notch1 receptor expression. To identify the cell lineages that receive Notch1 signal during heart development, we re-analyzed publicly available single-cell RNA-sequencing data from human embryos (post-conception weeks 6, 8, 12, and 19) and mouse embryos (embryonic days 12.5, 14.5, and 16.5). We found that endothelial cells expressed Notch1 in both species. To confirm Notch1 signaling rather than Notch1 receptor expression in heart development, we carried out transgenic mouse experiments that enabled tracing of both past and ongoing Notch1 signaling by combining the Notch1 receptor protein with the Gal4/UAS and Cre/loxP systems. Past Notch1 signal analysis confirmed that coronary endothelial cells, in addition to a small proportion of endocardial cells, received Notch1 signals during development, consistent with our findings in single-cell RNA-seq re-analysis. Ongoing Notch1 signaling was observed in the surface-covering cells of the vascular lumens, although its colocalization with an endothelial marker (Pecam1/Cd31) was unclear. Collectively, our findings suggested that coronary endothelial cells are Notch1 signal receivers, which should contribute to understanding the pathogenesis of congenital heart diseases.
Background:Vital pulp therapy (VPT) aims to preserve pulp vitality and stimulate regeneration of the dentin-pulp complex. Dental pulp stem cells (DPSCs) are clonogenic mesenchymal stem cells with high self-renewal and multipotent differentiation capacity. Bioactive cements such as mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM) are widely used as pulp-capping agents due to their biocompatibility. Platelet-rich fibrin (PRF), an autologous fibrin matrix rich in cytokines and growth factors, has been used to support tissue healing and regeneration. Methods:Human third-molar DPSCs were isolated and characterized, then cultured with MTA, CEM, and combinations of cement with PRF. Cell viability was assessed using the MTT assay, and expression of pluripotency markers (OCT4, SOX2, NANOG) was quantified by real-time PCR. Results:MTT assays showed that CEM alone significantly reduced DPSC viability compared to control, whereas MTA and MTA + PRF did not differ from control. Notably, co-treatment with PRF attenuated CEM's adverse effect. NANOG was significantly upregulated in the MTA and MTA + PRF groups relative to control, and SOX2 was significantly elevated in the MTA + PRF group. OCT4 remained unchanged across all conditions. Conclusion:The combination of MTA and PRF enhanced DPSC survival and upregulated pluripotency markers (NANOG, SOX2), supporting its potential application in regenerative VPT. In contrast, CEM alone had a detrimental effect on cell viability; however, this adverse effect was attenuated by PRF co-application. These findings suggest that MTA + PRF is a promising pulp-capping strategy for preserving DPSC function and promoting pulp regeneration.
Momordicae Semen (MS) is a traditional medicinal herb with reported antitumor potential. However, the impact of processing on its chemical composition and the underlying mechanisms against hepatocellular carcinoma (HCC) remain insufficiently understood. This study aimed to characterize processing-induced chemical changes and elucidate the bioactive constituents and molecular pathways responsible for improved anti-HCC effects. UPLC-QTOF/MS was used to compare chemical differences between crude and processed samples. Network pharmacology, molecular docking, and molecular dynamics (MD) simulations were applied to predict active compounds and core pathways. A DEN-induced rat HCC model was established to validate pathway regulation. Forty-one constituents were identified, with obvious chemical variations occurring after processing. A total of 207 disease-related targets were obtained, and Akt/mTOR/STAT3 served as the key pathway. The primary bioactive constituent, methyl gypsogenin 3-O-β-d-glucuronopyranoside (MG), exhibited favorable binding affinities toward the key proteins in the Akt/mTOR/STAT3 pathway, with calculated binding energies of -5.47 kcal/mol, -7.25 kcal/mol, and -5.71 kcal/mol, respectively. The 4RDD-MG complex maintained stable conformation during 100 ns MD simulation, with a Molecular Mechanics/Poisson-Boltzmann Surface Area (MM/PBSA) total binding free energy of -18.95 kcal/mol. In vivo results showed that high-dose MSF markedly alleviated liver injury and suppressed inflammatory cytokines (all P < 0.05), downregulated Ki67 expression, and significantly inhibited the phosphorylation of Akt, mTOR and STAT3, with superior efficacy compared with crude MS at the same dosage.
Diabetic retinopathy (DR), a major microvascular complication of diabetes, is a leading cause of vision loss. Neovascularization is a hallmark of DR, and circRNA-PSEN1 (circPSEN1) has emerged as a key regulator in its development. However, the precise molecular mechanisms by which circPSEN1 contributes to DR remain unclear. This study investigates the role of circPSEN1 in DR, focusing on its interaction with miR-150-5p and TRIM65. Human retinal microvascular endothelial cells (hRMECs) were cultured under high-glucose (HG) conditions to establish an in vitro DR model, and streptozotocin-induced DR rats were used for in vivo analysis. Lentivirus-mediated gene modulation was employed to alter expression levels. Cell viability, migration, angiogenesis, and inflammatory cytokine release were assessed using CCK-8, transwell, tube formation assays, and ELISA. Dual-luciferase reporter assays, RNA pull-down, and RNA immunoprecipitation (RIP) techniques were used to examine the interactions between circPSEN1, miR-150-5p, and TRIM65. Elevated circPSEN1 and TRIM65 levels, along with reduced miR-150-5p expression, were observed in retinal tissues from DR rats. In HG-stimulated hRMECs, circPSEN1 knockdown inhibited cell proliferation, migration, and angiogenesis, while reducing inflammatory cytokine release. These effects were reversed by miR-150-5p suppression or TRIM65 overexpression, suggesting a regulatory role of both molecules in DR. Mechanistically, circPSEN1 acts as a sponge for miR-150-5p, enhancing TRIM65 expression and contributing to DR pathogenesis. Our findings indicate that circPSEN1 may serve as a therapeutic target for DR.
Neem, or Azadirachta indica A. Juss., is a tree that is both versatile and native to the Indian subcontinent and Southeast Asia. It is frequently employed in traditional medicine to treat skin issues and pyrexia, as well as for environmental protection and insect control. This review reports the multifaceted potential of A. indica (neem), a plant that is profoundly rooted in traditional medicine. Neem has high value due to its rich composition of bioactive compounds, and in particular, has a potential for combating several pathogenic microbial communities. There is an increasing necessity to identify novel antimicrobial agents in light of the alarming increase in drug-resistant pathogens. Neem stands out as a promising option, showcasing a wide range of therapeutic benefits that go beyond just treating common issues like fever and skin problems. Its bioactive compounds boast impressive medicinal properties, offering anti-inflammatory, antimicrobial, and immunomodulatory effects that can be incredibly useful for tackling complex health conditions. This review emphasizes the importance of methods of bioactive extraction and biochemical features of neem leaf extracts. There are many bioactive substances such as, triterpenes, flavonoids, and phenolics that contribute to the plant's antibacterial, anti-inflammatory, and antioxidant properties. In addition, this review discusses the immunomodulatory effects and the influence of various extraction solvents on the yield and composition of bioactive compounds.
Radiosensitivity (RS) is an important biological factor that may contribute to future radiotherapy individualization. However, a comprehensive metric that adequately captures the complexities of radiation sensitivity has not yet been proposed, and the survival fraction at 2 Gy (SF2) remains the prevailing gold standard. Considerable variability in radiation responses across different cell types, together with heterogeneity within neoplastic and normal cell populations, continues to complicate radiosensitivity assessment.This study aimed to investigate the relationship between the radiosensitivity of malignant and non-malignant cells, with particular emphasis on cancer stem cells (CSCs). Blood samples and tumor biopsy specimens were collected from 20 breast cancer patients. Radiosensitivity was evaluated using clonogenic survival assays in lymphocytes, breast cancer cells, and breast cancer stem cells, while DNA damage and repair kinetics in lymphocytes were assessed using the γH2AX assay.The results showed that the geometric mean measured 30 min after radiation exposure, as well as the residual double-strand breaks (DSBs) at 3 and 24 h in the γH2AX assay, were significantly correlated with lymphocyte SF2. These findings suggest that the γH2AX assay may provide complementary information to clonogenic survival assay.However, no significant correlation was observed between the radiosensitivity of lymphocytes and that of tumor cells or CSCs. Breast cancer stem cells exhibited slightly higher survival following irradiation compared with the corresponding tumor cells, indicating a trend toward increased radioresistance.The observed radiosensitivity patterns may provide preliminary biological insights, but they should not be interpreted as direct clinical predictors without validation in larger cohorts with treatment outcome data. Further studies incorporating larger cohorts and additional biological indicators are required to improve the prediction of radiation response and clarify their potential relevance for future individualized radiotherapy research.
TRPA1 channels function as cold sensors and have been implicated in energy metabolism. We investigated the impact of TRPA1 on thermoregulation. Trpa1 -/- mice exhibited body weight loss and increased food intake when transitioning from 30°C to mild cold (14 days at 22°C), indicating increased energy metabolism in response to cold. Trpa1 -/- mice exhibited elevated oxygen consumption, while their core temperature remained unaffected. The BAT proteome in WT mice after 14 days at 22°C showed increased proteins related to glucose utilization and reduced proteins related to lipolysis, whereas Trpa1 -/- mice revealed increased PRDM16 and proteins associated with peroxisomal/mitochondrial β-oxidation. Transcripts of Cidea, Ucp1, Cpt1a, and Lcad were higher in Trpa1 -/- mice at 22°C compared to 30°C, relative to WT mice, with Prdm16 expression uniquely increased in Trpa1 -/- mice. Thermographic images revealed higher interscapular surface temperature in Trpa1 -/- mice exposed to 22°C. These data suggest that the TRPA1 channel reduces metabolic stress and contributes to thermogenic regulation in mice.
Colistin is increasingly used as last-line therapy for A. baumannii infections. This has led to the emergence of colistin-resistant strains of A. baumannii. Therefore, exploring novel approaches to fight colistin resistance in A. baumannii is urgently required. In our previous study, we reported that capsaicin combined with colistin could effectively circumvent colistin resistance in A. baumannii. Here, we employed untargeted metabolomics to explore the global metabolic perturbations of ATCC 19606-R induced by a combination of colistin (2 μg/mL) and capsaicin (32 μg/mL) compared to each of them acting alone at 1, 4, and 24 h.Data revealed that the metabolic changes of ATCC 19606-R were initially driven by capsaicin (at 1 h) and then by colistin (at 4 h). The combination treatment induced remarkable metabolic perturbations at 4 h, which were mainly caused by colistin. This metabolic impairment continued for 24 h. Results also showed that vital metabolic pathways, such as those involved in carbohydrate, nucleotide, and amino acid metabolism, underwent significant changes. On the other hand, the combination treatment had a negligible effect on lipid metabolism. Data in this study suggest that a combination of capsaicin and colistin is a promising candidate for treating A. baumannii infections.
The consumption of A2 milk, which contains only the A2 type of beta-casein, has been increasing because it is considered healthier. In contrast, milk that contains the A1 type of beta-casein may cause gastrointestinal discomfort due to the release of the peptide beta-casomorphin-7 during digestion. The milk is composed of valuable proteins, liposoluble vitamins, and mineral salts, that could be essential source of dietary lipids for humans. The lipidomic analysis allows the characterization of important milk lipids. In this context, the objective of this work was to describe and determine the lipid profile and differential compounds in genotypes A1A1, A2A2, and A1A2 for beta-casein from bovine milk. Lipid extraction followed by gas chromatography coupled with mass spectrometry were performed on 54 bovine milk samples. As results, 12 differential metabolites were found, where the cholesterol; Cholesta-3,5-diene and Cholest-5-ene-3-ol(3. beta.)-carbonochloridate were the most abundant lipid compound in A2 milk. The identified lipids participate of the important pathways, such as the fatty acyl. The data obtained in this exploratory study provide insights into the main differences between the lipidomic profile of bovine milk present different beta-caseins in its composition.
Osteoclasts (OCs) play an important role in bone remodeling and bone resorption. Furthermore, OCs are considered to initiate the process of osteoinduction by stimulating the osteogenic differentiation of mesenchymal stromal cells (MSCs). OCs are large, multi-nucleated cells that originate from mononuclear precursors via cell fusion. Despite progress in understanding the molecular pathways involved, the dynamic morphological changes that precede and accompany fusion remain poorly defined. This study aims to characterize the spatiotemporal morphodynamics of osteoclast precursor cells (OCPs) during RANKL-stimulated differentiation, and identify structural features associated with fusion potential. Murine macrophages (RAW264.7 cells) were used as an in vitro model to study osteoclastogenesis. OCPs fusion and formation were induced by using 50 ng/mL receptor activator of nuclear factor kappa-B ligand (RANKL). Time-lapse holotomographic live-cell recordings were performed immediately after RANKL addition over 72 h using a Nanolive 3D cell explorer-fluo microscope, enabling continuous, label-free visualization of individual cell dynamics. Quantitative morphometric analysis was conducted using refractive index (RI)-based measurements with EVE analysis software to assess parameters including form factor, eccentricity, compactness, and cell extent. Finally, OCPs underwent progressive morphological remodeling prior to fusion, exhibiting increased elongation and shape irregularity. Multiple fusion phases were captured, including mono-mononuclear, mono-multinuclear, and multi-multinuclear fusion events. Notably, tunneling nanotubes-like membrane tethers (TNTs) frequently appeared before and during fusion and were associated with mediate directional movement and intercellular contact. Mononuclear cells displaying larger and more irregular morphology were more likely to participate in TNT-mediated fusion with multinucleated osteoclasts. Our findings demonstrate the predictive value of cell morphology in osteoclastogenesis and suggest that TNTs facilitate fusion coordination. This dynamic, single-cell-level perspective offers new insights into the morphodynamic characteristics of osteoclastogenesis and supports the notion that modulation of cell morphology may serve as a potential strategy to regulate osteoclast differentiation and function.
Tasisulam, an acyl-sulfonamide compound, is being investigated in clinical trials for the treatment of several malignancies, including non-small cell lung cancer, lymphoma, breast cancer, melanoma, ovarian cancer, colon cancer, and other solid tumors, by promoting apoptosis. However, anemia is among the adverse consequences of tasisulam therapy and is potentially caused by increased eryptosis or premature erythrocyte senescence, characterized by cell contraction and phosphatidylserine (PS) translocation. Underlying signals associated with eryptosis include increased intercellular calcium activity ([Ca2+]i), oxidative stress, excess ceramide production, and stimulation of various kinases (protein kinase C, p38 kinase, casein kinase-1, etc.) or caspases. This research investigated the potential of tasisulam to induce eryptosis and its underlying signaling pathways. Human erythrocytes (0.4%) were incubated with 75, 150, or 300 μg/ml tasisulam for 48 h at 37°C. Flow cytometry revealed that tasisulam (≥300 μg/ml) significantly increased erythrocyte apoptosis, [Ca2+]i, reactive oxygen species (ROS), and ceramide formation without causing cell membrane shrinkage. The effect of tasisulam on erythrocyte death was significantly reduced by the removal of extracellular calcium or the inhibition of casein kinase. In conclusion, tasisulam triggers eryptosis by stimulating calcium influx, ceramide generation, oxidative stress, and casein kinase 1 activation, which may be associated with tasisulam-associated anemia.
Background Keloid is a pathologic scar formation resulting from trauma, burns, surgery, or unknown causes, leading to deformity and varying degrees of functional impairment, significantly impacting the physical and mental well-being as well as the quality of life of affected individuals. Studies have indicated that Mesenchymal Stem Cells (MSCs) can effectively promote wound healing in various injuries, reducing the formation of keloid. This study aims to explore an in vitro model of keloid and investigates the effects of exosomes from human Umbilical Cord MSCs (HUCMSC) on fibroblast proliferation and apoptosis in keloid model. Methods Methods include isolating primary fibroblasts, extracting HUCMSC exosomes (HUCMSC-Exos), and establishing a histamine-induced keloid model in vitro. Results show dose-dependent inhibition of keloid fibroblast activity by HUCMSC-Exos, affecting proliferation, apoptosis, and cell cycle. Through gene sequencing analysis, we investigated the abnormal gene expression and signaling pathway changes of histamine-induced keloid fibroblast before and after co-cultivation with HUCMSC-Exos. Results It turns out that HUCMSCs-Exos effectively inhibit the proliferation of keloid fibroblasts, promoting apoptosis and exhibiting a dose-dependent effect. We also proved that HUCMSCs-Exos regulated genes and inhibit signaling pathway by upregulating the expression of intracellular genes, ultimately inhibiting keloid fibroblasts proliferation activity and enhancing apoptosis. The research found that HUCMSC-Exos as potential regulators of keloid fibroblast behavior, offering insights for keloid treatment strategies.
Adipose-derived stem/stromal cells (ASCs) are promising candidates for bone tissue engineering due to their abundance, accessibility, and osteogenic differentiation capacity. However, substantial inter-donor variability limits their standardized clinical application. This study investigated whether metabolic characteristics and donor-related parameters are associated with ASC osteogenic differentiation potential. ASCs were isolated from human subcutaneous adipose tissue (n = 26) and cultured under osteogenic conditions. Cell viability was evaluated at days 2 and 14, including cell proliferation (crystal violet staining), overall metabolic activity (resazurin conversion), as well as glucose uptake (2-NBDG) and mitochondrial membrane potential (TMRE). Osteogenesis was quantified by alkaline phosphatase activity, calcium deposition, and secretion of SPARC and osteocalcin (BGLAP). Osteogenic induction significantly enhanced osteogenic markers and mineralization across donors and was accompanied by a distinct metabolic phenotype characterized by reduced per-cell metabolic activity and glucose uptake, and elevated mitochondrial membrane potential. Early (day 2) metabolic parameters did not predict subsequent mineralization capacity, whereas metabolic activity at day 14 correlated inversely with calcium deposition. Donor age, body mass index, and blood glucose levels were not associated with osteogenic outcomes. These findings indicated that inter-donor variability in ASC osteogenesis was accompanied by a characteristic late-stage metabolic signature and suggested that metabolic profiling may support functional characterization of osteogenic differentiation potential.
