Search Results (2,192)

Scaffold architecture is a key determinant of cell behavior and tissue regeneration in bone tissue engineering, yet the influence of pore size under dynamic culture conditions remains incompletely understood. This study aimed to evaluate the effects of scaffold pore size on osteogenic differentiation of porcine bone marrow-derived mesenchymal stem cells (pBMSCs) cultured in a rotational oxygen-permeable bioreactor system (ROBS). Three-dimensionally (3D) printed beta-tricalcium phosphate (β-TCP) scaffolds with pore sizes of 500 µm and 1000 µm were seeded with pBMSC and cultured for 7 and 14 days under dynamic perfusion conditions. Gene expression analysis revealed significantly higher levels of osteogenic markers (Runx2, BMP-2, ALP, Osx, Col1A1) in the 1000 µm group, particularly at the early time point, with the later-stage marker Osteocalcin (Ocl) rising faster and higher in the 1000 µm group, after a lower expression at 7 days. ALP activity assays corroborated these findings. Despite having lower mechanical strength, the 1000 µm scaffolds supported a homogeneous cell distribution and high viability across all regions. These results suggest that larger pore sizes enhance early osteogenic commitment by improving nutrient transport and fluid flow in dynamic culture. These findings also support the use of larger-pore scaffolds in bioreactor-based preconditioning strategies and underscore the clinical importance of promoting early osteogenic differentiation to reduce in vitro culture time, an essential consideration for the timely preparation of implantable grafts in bone tissue engineering. Full article

A complex and gradual process, the epithelial–mesenchymal transition (EMT) occurs both during embryonic development and tumor progression. Cells undergo a transition from an epithelial to a mesenchymal state throughout this process. More and more evidence points to EMT as a cause of increased metastatic spread of prostate cancer (PCa), along with stemness enhancement and therapy resistance. Here, we used bioinformatic methods to analyze gene expression microarray data, single-cell RNA sequencing, oncogenes, and tumor suppressor genes (TSGs) in order to reconstruct the network of differentially expressed genes (DEGs) involved in the epithelial–mesenchymal transition with PCa. No prior study has documented this sort of analysis. We next validated our results using data from the Cancer Genome Atlas (TCGA), which included microarray and single-cell RNA sequencing. Potentially useful in PCa diagnosis and treatment are extracellular matrix in epithelial–mesenchymal transition genes, including ITGBL1, DSC3, COL4A6, ANGPT1, ARMCX1, MICAL2, and EPHA5. In this study, we aimed to shed light on the molecular characteristics and pathways of DEGs in PCa, as well as to identify possible biomarkers that are important in the development and advancement of this cancer. These insights have important implications for understanding prostate cancer progression and for the development of therapeutic strategies targeting ECM-mediated pathways. Full article

Drought stress is a major abiotic constraint that severely restricts the growth of Medicago falcata L. by inducing the accumulation of reactive oxygen species (ROS) in plants. WRKY transcription factors (TFs) play a key role in regulating plant responses to drought stress. In this study, we investigated the role of the MfWRKY40 gene in drought tolerance. Under mannitol and ABA stress treatments, MfWRKY40-overexpressing lines (OEs) showed significantly longer primary roots, increased lateral roots, and higher fresh weight compared to wild-type (Col) lines, indicating significantly enhanced growth and drought tolerance. Similarly, under soil drought conditions, transgenic Arabidopsis thaliana exhibited enhanced drought tolerance. NBT staining demonstrated decreased ROS accumulation in transgenic lines after stress treatment. Correspondingly, the MfWRKY40-overexpressing lines displayed significantly lower levels of hydrogen peroxide (H₂O₂), superoxide anion (O₂⁻), and malondialdehyde (MDA) compared to Col, along with elevated activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), as well as increased proline (Pro) content. Furthermore, MfWRKY40 upregulated the expression of antioxidant enzyme genes (AtPOD3, AtSOD4, and AtCAT1) and modulated the expression of other drought-related genes. In summary, our results demonstrate that MfWRKY40 enhances drought tolerance in A. thaliana by improving ROS scavenging capacity. This study provides a theoretical foundation for further exploration of MfWRKY40’s functional mechanisms in drought stress adaptation. Full article

Background/Objectives: Cancer-associated fibroblasts (CAFs) play a pivotal role in the tumor microenvironment. We conducted an analysis using RNA sequencing to identify specific markers for CAFs compared to normal fibroblasts (NFs) in non-small-cell carcinoma (NSCLC). Methods: CAFs and NFs were isolated and cultured from tumor tissues (primary tumor or metastatic lymph nodes) and matched non-tumor tissues, respectively. Bulk RNA sequencing was conducted on isolated CAFs and normal fibroblast NFs. Differential expressions, gene set enrichment, and CAF subpopulation prediction analyses were performed. Results: During the study period, 27 CAFs and 12 NFs were isolated and cultured from tumor and non-tumor tissues in patients with treatment-naïve NSCLC. Among them, 22 CAFs and 11 NFs were included in the RNA sequencing analysis. The 22 CAF samples consisted of 12 adenocarcinomas and 10 squamous cell carcinomas (SqCC), with 16 samples from the lungs and 6 samples from the lymph nodes. Notably, COL11A1, GREM1, CD36, and GAS6 showed a higher expression in CAFs than in NFs, whereas TNC and CXCL2 were more abundantly expressed in NFs. CD36 levels were elevated in CAFs from lymph nodes (LN-CAFs) compared with those from lung specimens (Lung-CAFs) and NFs. COL11A1 levels in Lung-CAFs surpassed those in LN-CAFs and NFs. Both GREM1 and GAS6 showed a strong expression in Lung-CAFs and LN-CAFs relative to NFs. CAFs exhibited features of the myofibroblast CAF subpopulation, whereas NFs displayed traits of the antigen-presenting CAF subtype. In the co-culture model of CAFs and THP-1 cells, the knockdown of GREM1 or GAS6 in CAFs significantly decreased the M2 marker expression in macrophages. Conclusions: In NSCLC, GREM1 and GAS6 can be valuable diagnostic targets for CAFs from primary tumors and metastatic sites; they warrant further study. Full article

The electrochemical oxidation of anodic metal (cobalt, nickel, zinc or cadmium) in a cell containing an acetonitrile solution of the ligand (E)-N-(2-(((2-hydroxynaphthalen-1-yl)methylene)amino)phenyl)-4-methylbenzenesulphonamide (H₂L) affords complexes with the general formula [ML] (M = Co, Ni, Zn and Cd). Additionally, it was possible to obtain complexes with the general formula [MLL′] when L′ = 2,2-bipyridine (2,2-bpy), 4,4-bipyridine (4-4′-bpy) or 1,10-phenanthroline (phen) was present in the electrolytic cell. All of the compounds obtained have been characterized via microanalysis, IR spectroscopy, mass spectrometry, UV–visible spectroscopy and, in the case of diamagnetic compounds, via ¹H NMR spectroscopy. Further structural and electronic characteristics of these adducts have been obtained via DFT simulations. The compounds NEt₄[CoL₂] (1), [NiL(H₂O)] (2), [NiL(CH₃CN)(H₂O)]₂ (3), [Ni₂L₂(4,4′-bpy)] (4), [Zn₂L₂(MeOH)₂] (5) and [ZnL(2,2′-bpy)](CH₃CN) (6) have been characterized via X-ray diffraction. In this paper, we present a detailed study of the different behavior of the above-mentioned ligand depending on the metal and/or the presence of ancillary ligands. Full article

The present study is aimed at investigating the effects of grazing on the meat quality of Tibetan sheep, as well as the associated molecular mechanisms. A total of ten Tibetan sheep were utilized and equally allocated into two groups: grazing and pen-feeding. To assess the intramuscular fat (IMF) content, Soxhlet extraction was performed on the biceps femoris muscle. Additionally, transcriptome sequencing was carried out to evaluate the expression profiles of RNAs, facilitating the construction of a ceRNA regulatory network. The results demonstrated that the IMF content in the grazing group was significantly higher compared to the pen-feeding group, implying that grazing might foster the formation of Type I muscle fibers, thereby enhancing meat quality. Moreover, the expression levels of circRNAs, such as novel_circ_001331, novel_circ_012918, novel_circ_029843, and novel_circ_059962, were markedly up-regulated in the grazing group. These circRNAs may alleviate the inhibitory effects on genes like COL8A1, MYLK3, and NOX4 by interacting with miR-381-y, miR-7144-x, miR-16-z, miR-8159-x, novel-m0040-3p, novel-m0092-5p, and oar-miR-329a-3p. These circRNAs and miRNAs are predominantly involved in the MAPK, Wnt, and VEGF signaling pathways and could be implicated in biological processes such as muscle fiber type switching and energy metabolism. This research offers valuable insights for improving the meat quality of Tibetan sheep and provides a foundation for exploring the role of circRNA and miRNA in the regulation of meat quality under grazing conditions. Full article

Background: Cardiomyocyte death is a key factor in myocardial ischemia–reperfusion injury (MI/RI), and the expression patterns and molecular mechanisms of pyroptosis-related genes (PRGs) in ischemia–reperfusion injury are poorly understood. Methods: The mouse MI/RI injury-related datasets GSE61592 and GSE160516 were obtained from the Gene Expression Omnibus database, and differential expression analysis was performed on each to identify differentially expressed genes (DEGs). The DEGs were intersected with the PRGs obtained from GeneCards to identify differentially expressed PRGs in MI/RI. Enrichment analysis identified key pathways, while PPI network analysis revealed hub genes. The expression patterns and immune cell infiltration of hub genes were also investigated. The molecular docking prediction of key genes was performed using MOE software in conjunction with the ZINC small molecular compounds database. Key gene expression was validated in an external dataset (GSE4105), a mouse MI/RI model, and an HL-1 cell hypoxia/reoxygenation model via RT-qPCR. Results: A total of 29 differentially expressed PRGs were identified, which are primarily associated with pathways such as “immune system process”, “response to stress”, “identical protein binding”, and “extracellular region”. Seven key genes (Fkbp10, Apoe, Col1a2, Ppic, Tlr2, Fstl1, Serpinh1) were screened, all strongly correlated with immune infiltration. Seven FDA-approved small molecule compounds exhibiting the highest docking potential with each key gene were selected based on a comprehensive evaluation of S-scores and hydrogen bond binding energies. Apoe, Tlr2, and Serpinh1 were successfully validated across external datasets, the mouse MI/RI model, and the cardiomyocyte H/R model. Conclusions: Apoe, Tlr2, and Serpinh1 may be key genes involved in MI/RI-related pyroptosis. Targeting these genes may provide new insights into the treatment of MI/RI. Full article

Stauntonia hexaphylla (Thunb.) Decne (SH), a medicinal plant from the Lardizabalaceae family, holds traditional importance in East Asia for treating rheumatism. Steam treatment is commonly applied to enhance its medicinal properties, but the chemical and biological changes resulting from this process remain unexplored. This study compared steamed and untreated SH fruit (SHF) extracts, analyzing their chemical composition, antioxidant activity, and effects on bone health using in vitro models. Steamed SHF extracts exhibited increased levels of 5-hydroxymethylfurfural (5-HMF), total flavonoids, phenolics, and enhanced antioxidant activity. Bone health assessment using osteoclasts differentiated from RAW 264.7 cells and osteoblasts from MC3T3-E1 cells revealed that steamed extracts promoted alkaline phosphatase activity, calcium nodule formation, and collagen synthesis in osteoblasts while inhibiting tartrate-resistant acid phosphatase (TRAP) activity in osteoclasts. Additionally, steamed SHF extracts effectively modulated gene expression related to osteoclastogenesis and osteoblastogenesis by downregulating TRAP, NFTAc1, RANK, MMP9, c-Fos, and TRAF6 while upregulating ALP, Runx2, BGLAP, Col1a1, and OPG. The component 5-HMF played a pivotal role in promoting alkaline phosphatase and inhibiting TRAP activities. These findings suggest that steamed SHF may offer a promising therapeutic approach for postmenopausal osteoporosis. Full article

This study addresses the role of cytosine methylation in the fine-tuning of flowering time under water deficit in Arabidopsis thaliana. A drm1 drm2 cmt3 (ddc) triple methylation mutant was used together with the Col-0 wild type. The plants were grown under long-day conditions with water deficit induced by cessation of watering starting 12 days after seeding. Col-0 showed a 1-day delay in flowering as a result of the treatment. In contrast, ddc showed a 2-day delay regardless of the experimental conditions. We found that the two b-box domain proteins, BBX16/COL7 and BBX17/COL8, became overexpressed in the ddc background and in Col-0 under water deficit 24 days after seeding. Additionally, the NF-YA2 transcription factor became correspondingly down-regulated. Our results support a model where BBX16/COL7 and BBX17/COL8 interact with CONSTANS to delay the induction of FT under long-day conditions. NF-YA2, which is also recognized as a promoter of FT expression, with its down-regulation causes additional delay of FT-induced flowering. The plants overcome the BBX/NF-YA inhibition easily, resulting in a relatively small delay in flowering. The expression patterns of the three genes suggest the involvement of cytosine methylation in their regulation; however, no differential methylation could be found in cis that can explain these effects. The results therefore suggest a trans acting mechanism. Considering that the activities of BBX16/COL7 and BBX17/COL8 in different physiological conditions are not elucidated, this paper provides a background for future experiments targeting the role of these genes in the fine-tuning of flowering time in A. thaliana. Full article

The glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) are ligand-activated transcription factors that regulate epidermal homeostasis, inflammation, and function. Prior studies using epidermal-specific conditional single and double knockout mice have shown their importance in skin physiology; however, clinically human disease is largely treated pharmacologically. Our objective was to examine how systemic MR/GR antagonism affects cutaneous gene expression and epidermal thickness in aged (18-month-old) C57BL/6J female mice. Mice were treated with selective GR (relacorilant), selective MR (eplerenone), or dual GR/MR (miricorilant) antagonists for 8 weeks. Quantitative RT-qPCR analysis of the skin showed that miricorilant significantly upregulated Sgk1, a GR/MR target. Miricorilant also increased the expression of keratinocyte differentiation markers and downregulated key inflammatory cytokines and Col3a1, a collagen subtype associated with tissue remodeling. Relacorilant suppressed Scnn1g, a subunit of the epithelial sodium channel. None of the antagonists significantly altered proliferation markers, epidermal thickness, or regulators of glucocorticoid activity. Our findings show that miricorilant downregulated inflammatory cytokines and increased differentiation marker expression without affecting epidermal thickness, suggesting its potential to treat inflammatory skin diseases. The results contrast with data from GR/MR knockout studies, highlighting the likely significance of receptor dynamics. Further studies of antagonist effects on receptor interactions with co-regulators appear warranted. Full article

Seed size is a key trait affecting evolution and agronomic performance by influencing seedling establishment in natural populations and crop yields. The Arabidopsis thaliana Seed Size QTL1 (SSQ1) locus explains 10–15% of the variation in seed size. We report here that the causal gene for this locus is Tetratricopeptide Repeat Protein 2 (TPR2), which encodes a co-chaperone. Expressing TPR2 across ecotypes and genotypes showed consistent dosage effects. Each additional TPR2^Col-0 allele increased seed mass and volume by 10–14% with high reliability in Col-0, Sha, Tsu-1, and tsu2 genetic backgrounds. Reciprocal genetic crosses indicated that this locus acts maternally, consistent with female sporophytic or female gametophytic mutations. To elucidate how TPR2 regulates seed size, the biomass composition of seeds was measured. While oil content remained unchanged, sucrose levels were markedly elevated in TPR2^Col-0 transformant lines and reduced in tpr2 mutants. Interestingly, heterologous expression of TPR2^Col-0 across genetic backgrounds increased seed protein accumulation by 18% on average. Based on these changes in sucrose and protein levels, potential modes of action for TPR2 are discussed. Full article

Abscisic acid (ABA) is a crucial phytohormone that functions as a master regulator of plant growth and development, as well as responses to diverse abiotic stresses, by integrating environmental cues with developmental programs. The transcription factor ABA INSENSITIVE 5 (ABI5) functions at the central hub of the ABA signaling pathway and mediates the expression of its target genes. Emerging evidence reveals extensive crosstalk between calcium-dependent protein kinases (CPKs)-mediated calcium signaling and the ABA-ABI5 cascade, enabling plants to balance growth and stress responses. However, the molecular mechanisms underlying the interactions between CPKs and ABA-ABI5 signaling are still elusive. In this study, we revealed that CPK2 enhances sensitivity to ABA during both seed germination and seedling root growth by promoting ABA-induced ABI5 expression and increasing ABA-mediated ABI5 stability. Compared to the wildtype (Col-0), the CPK2-OE line exhibited the highest sensitivity to ABA in both seed germination and root growth, while the cpk2abi5-7 double mutant showed the least sensitivity. The single mutants cpk2 and abi5-7, as well as the abi5-7CPK2-OE2 line, displayed intermediate phenotypes, suggesting that CPK2 acts upstream of ABI5. Biochemical and molecular biological studies revealed that CPK2 physically interacts with ABI5 and directly phosphorylates it at Ser42, Ser145, and Thr201. Moreover, both ABA-induced ABI5 expression and protein accumulation were significantly reduced in cpk2 mutants upon ABA treatment. Taken together, these findings provide compelling evidence that CPK2 exacerbates the ABA inhibition of seed germination and root growth by enhancing both the expression and stability of ABI5, thereby reinforcing stress adaptation during early plant development. Full article

Background: Early detection and prevention of colorectal cancer (CRC) are key goals of population-based screening. Several diagnostic tests have been proposed for CRC screening. This study compares the diagnostic accuracy of colon capsule endoscopy (CCE), colonoscopy (COL) and computed tomographic colonography (CTC), focusing on risk factors such as polyps. Methods: We conducted a systematic review with meta-analyses and network meta-analysis. Pooled estimates of sensitivity (SE), specificity (SP), positive predictive value (PPV) and negative predictive value (NPV) were calculated using a random-effects model. Diagnostic performance was assessed for first- and second-level screening based on effect size estimates. Results: For first-level screening, sensitivity was 0.79 (95% CI: 0.60–0.91) and specificity 0.95 (95% CI: 0.88–0.98); PPV and NPV were 0.89 and 0.97, respectively. In second-level screening, sensitivity was 0.75 (95% CI: 0.65–0.83), specificity 0.95 (95% CI: 0.92–0.97), PPV 0.76 and NPV 0.95. The indirect sensitivity estimate of CCE vs. COL (SMD = 0.30; 95% CI: 0.12–0.47) was lower than the direct estimate for CTC (SMD = 0.44; 95% CI: 0.29–0.59). CCE showed better comparative performance than CTC relative to COL (CCE SMD = −0.18; 95% CI: −0.29 to −0.06 vs. CTC SMD = −0.98; 95% CI: −1.07 to −0.90). However, both CCE and CTC had lower specificity than COL. Conclusions: CCE represents a valuable tool for early CRC detection. Test selection should be guided by clinical and epidemiological settings to optimize screening strategies. Full article

The interplay between angiogenesis and the immune system is intricate, with the potential to either enhance or repress the immune response. Angiogenesis-related genes (ARGs) are significant for the development, growth, and immune response of tumors. Understanding their prognostic significance and molecular characteristics in skin melanoma can guide and refine therapeutic strategies. Here, we analyzed the TCGA-SKCM dataset and explored the ARG expression between skin melanoma and normal skin, as well as between primary and metastatic tumors. Kaplan–Meier analyses were conducted to assess the overall, disease-specific, and progression-free survival. Additionally, comprehensive immune profiling was carried out utilizing advanced bioinformatics tools to evaluate immune checkpoint gene expression and immune cell infiltration. Our findings highlighted strong prognostic associations for S100A4, ITGAV, and COL3A1. Molecular characterization showed a significant upregulation of PTK2, CXCL6, COL3A1, COL5A2, PF4, TNFRSF21, LRPAP1, VTN, TIMP1, SPP1, and OLR1 in SKCM compared to that in normal skin. Immune analyses, including Immune Checkpoint Gene Analysis, Immune Infiltration Analysis, Immune Cell Analysis, and Immune Cell Profiling, demonstrated both positive and negative correlations between ARGs expression and immune cell infiltration, emphasizing the multifaceted role of these genes in immune modulation. The study underscores the prognostic relevance of ARGs in skin melanoma and their contribution to tumor immunity. Overall, our findings expand our understanding of melanoma immunogenetics, suggesting the use of angiogenesis-related genes not merely as vascular regulators, but also as immune modulators. Full article

Skin wound healing and barrier restoration are complex, tightly regulated processes critical for maintaining skin integrity, particularly in aged or compromised skin. This study investigated the wound healing efficacy and epidermal barrier-restoring effects of ε-Viniferin, a bioactive resveratrol dimer, and Vino Chocolate™, a grape flower-derived extract from Vitis labruscana ‘Campbell Early’ cell cultures enriched with ε-Viniferin. An HPLC analysis confirmed a high concentration of ε-Viniferin (547.58 ppm) in the cell culture-derived extract. In vitro assays conducted on HaCaT keratinocytes and HDFn fibroblasts demonstrated that the treatment with ε-Viniferin and Vino Chocolate™ significantly enhanced fibroblast migration. ELISA analyses showed that both treatments induced a dose-dependent increase in pro-collagen type I (COL1A1), with ε-Viniferin at 1 ppm demonstrating superior efficacy compared to TGF-β1. Additionally, these compounds notably suppressed the expression of matrix metalloproteinases MMP-1 and MMP-3, displaying effects comparable to or greater than retinoic acid. The Western blot analysis further revealed an increased filaggrin expression in keratinocytes, suggesting an improved epidermal barrier function. Collectively, these results indicate that ε-Viniferin and Vino Chocolate™ effectively promote extracellular matrix remodeling, modulate inflammatory responses, and enhance epidermal barrier integrity. These findings highlight their potential as multifunctional bioactive agents for cosmeceutical applications and emphasize the advantages of plant cell culture technology as a sustainable, innovative platform for advanced skincare ingredient development. Full article