Disrupted sleep from obstructive sleep apnea or insomnia promotes an inflammatory environment. Studies in mice have shown that experimental sleep fragmentation (SF) increases pro-inflammatory cytokine gene expression in the brain and peripheral tissues as well as increases circulating glucocorticoids. Although glucocorticoids classically exert anti-inflammatory effects at pharmacological concentrations, they may also modulate immune function in a context- and dose-dependent manner. To investigate whether glucocorticoids regulate pro-inflammatory responses, we used a classical adrenalectomy (ADX) ablation and corticosterone (CORT) replacement design. Adult male C57BL/6J mice received bilateral ADX, sham surgery, or ADX with CORT replacement in drinking water and were exposed to acute (24 h) SF or no SF (NSF). Pro-inflammatory gene expression (interleukin-1 beta and tumor necrosis factor-alpha) was quantified in the hippocampus, hypothalamus, and prefrontal cortex, as well as peripheral tissues including the liver, spleen, heart, and epididymal white adipose tissue. ADX effectively reduced serum CORT, while CORT replacement produced pharmacological hormone levels. Pharmacological CORT treatment robustly suppressed pro-inflammatory gene expression across multiple brain regions (prefrontal cortex, hypothalamus) and peripheral tissues (liver, heart, spleen) compared with ADX and/or sham mice, indicating a strong anti-inflammatory effect at elevated glucocorticoid concentrations. In contrast, ADX mice showed modest, tissue-specific increases in inflammatory gene expression, primarily in the liver and heart, suggesting limited regulation by physiological CORT levels. SF did not significantly increase pro-inflammatory gene expression compared with controls. In conclusion, CORT exerts potent, dose-dependent, and tissue-specific suppression of pro-inflammatory gene expression that is not modulated by acute SF in male mice.

microRNAs (miRNAs) regulate gene expression post-transcriptionally and play essential roles in various cellular processes. Disruption of their normal expression contributes to the pathogenesis of numerous diseases, including cancer, diabetes, depression, and neurological disorders. Consequently, restoring or inhibiting miRNA function has become a major focus of research, utilizing advanced technologies. Despite their potential, the clinical application of these technologies remains limited due to challenges related to delivery, specificity, cost, and safety. In contrast, growing evidence suggests that natural compounds-particularly flavonoids, a class of polyphenolic compounds abundant in plant-based foods such as fruits, vegetables, tea, and whole grains-can beneficially modulate miRNA expression. Studies across various disease models have demonstrated that flavonoids influence disease-related pathways through miRNA regulation, showing promising results in reducing disease complications, especially in preclinical mouse models. Therefore, diets rich in flavonoids are not only beneficial for disease prevention but also hold potential therapeutic value through the targeted modulation of miRNAs. The effectiveness of flavonoids across a wide spectrum of chronic diseases introduces a novel class of miRNA modulators with significant potential for clinical application. This review comprehensively examines recent studies on the use of various flavonoids in cellular models, animal models, and human tissue samples related to cancer, diabetes, depression, and neurological disorders. It further explores the molecular mechanisms through which flavonoids regulate miRNA expression, including transcriptional control, epigenetic modifications, and modulation of signaling pathways. Additionally, the review discusses the therapeutic opportunities and challenges associated with flavonoid use and proposes potential strategies to overcome these limitations.

Root immunity is vital for defending plants against soilborne pathogens, yet the regulatory mechanisms underlying this process remain incompletely understood. Here, we characterize a novel regulatory module comprising the long non-coding RNA LASSO, microRNA miR169f, and the transcription factor NF-YA1 that regulates rice root immunity. miR169f overexpression lines were generated and investigated using pathogen susceptibility assays, transcriptome analysis (mRNA-seq), and hormone measurements. Target genes were validated using 5' RLM-RACE (RNA Ligase-Mediated Rapid Amplification of cDNA Ends), degradome sequencing, transient co-expression in Nicotiana benthamiana, and RT-qPCR was used to validate and extend the transcriptome dataset. miR169f-overexpression plants exhibit reduced susceptibility to nematodes and the oomycete Pythium arrhenomanes. On the contrary, lines with reduced miR169f expression (STTM169f) as well as LASSO- and NF-YA1-overexpression lines show enhanced susceptibility. LASSO is rapidly induced in roots in response to nematode secretions, and most likely acts as a target mimic that sequesters miR169f, thereby relieving repression of NF-YA1. The miR169f fine-tunes defence responses by influencing the jasmonic acid (JA) pathway under basal conditions while enabling strong JA-responsive gene activation during nematode infection. This dynamic regulation links post-transcriptional control with hormone signalling to shape root immunity. Together, our results reveal a previously uncharacterized non-coding RNA network that orchestrates root defence responses.

Claudin-1 (CLDN1), a tight junction protein overexpressed and mis-localized in colorectal cancer (CRC), plays a critical role in tumor progression, stemness, and therapy resistance. Integrative analyses of bulk and single-cell transcriptomic datasets revealed that CLDN1 is enriched in stem-like CRC cells, increases during metastatic progression, and is associated with microsatellite stable disease. High CLDN1 expression correlates with epithelial-to-mesenchymal transition and activation of oncogenic pathways including AKT/mTOR, Myc, and NF-κB. Here, we evaluate the therapeutic efficacy of an investigational humanized monoclonal antibody (H3L3) directed against overexpressed non-junctional CLDN1 in preclinical models of CRC. In xenograft mouse models and patient-derived organoids (PDOs), CLDN1 mAb significantly reduced tumor growth in CLDN1-expressing tumors. Mechanistically, CLDN1 mAb disrupted CLDN1-mediated signaling, notably inhibiting the AKT/mTOR pathway. CRISPR-mediated CLDN1 knockout abolished H3L3 efficacy, confirming target specificity. Transcriptomic analysis of PDOs treated with H3L3 revealed broad suppression of stemness, oncogenic signaling, and hallmark cancer pathways. Single-cell analysis further demonstrated that targeting CLDN1 modulates cellular plasticity, driving stem-like tumor cells toward a more differentiated epithelial phenotype. Together, these findings establish anti-CLDN1 monoclonal antibodies as a promising therapeutic approach for CRC. The selective binding of CLDN1 mAb to tumor-associated exposed and overexpressed CLDN1, along with its capacity to inhibit key oncogenic pathways and reprogram cancer cell states including tumors with different mutations conferring resistance to standard of care, underscores its potential to improve treatment outcomes in patients with CLDN1-expressing CRC.

Chronic hepatitis B virus (HBV) infection poses a significant global public health challenge. Current therapies rarely achieve a functional cure, defined as the clearance of hepatitis B surface antigen (HBsAg) and fulfillment of other criteria. A critical unmet need is the development of agents that effectively suppress HBsAg production. Notably, serum HBsAg in patients predominantly originates from subviral particles (SVPs). To identify compounds inhibiting SVP production, we previously developed a cell model (HepG2-S-HiBiT) secreting HiBiT-tagged HBsAg, enabling high-throughput screening. Using this model, we screened a library comprising more than 5000 compounds and identified a hit compound (Compound 2) that potently inhibited HBsAg production. Subsequent structural optimization yielded a lead derivative, designated D4. In vitro cellular assays confirmed that D4 suppresses HBsAg production through a transcription-dependent mechanism. Transcriptomic profiling further revealed that Transmembrane Protein 40 (TMEM40) is significantly upregulated following D4 exposure. Further mechanistic investigations established that TMEM40 exerts anti-HBV activity via activation of the JAK-STAT signaling pathway. Collectively, our findings demonstrate that D4 represents a promising lead scaffold for the development of novel anti-HBV therapeutics, exerting its antiviral effects by upregulating TMEM40 and subsequently activating the JAK-STAT pathway to suppress viral transcription.

Resveratrol is a polyphenolic compound with reported anti-aging properties that exhibit complex and context-dependent biological effects, while its potential multigenerational influence remains unclear.

Brassinolide (BR) plays a pivotal role in regulating plant secondary metabolism and can promote the accumulation of bioactive compounds. However, relatively few studies have explored the application of BR in Salvia miltiorrhiza, and the mechanistic basis for its function in this context remains poorly understood.

Axial elongation and cell fate decisions during early embryonic development are regulated by gradients of signaling molecules, such as retinoic acid (RA) and fibroblast growth factor (FGF). To assess the role of RA in vitro, studies have relied on its direct addition to the medium, which can produce teratogenic effects. Here, we examined the role of a RA precursor, retinol, on axial elongation, anteroposterior development, and FGF-dependent signaling in murine gastruloids, an in vitro model system for early embryogenesis. Rather than applying RA ectopically, we supplemented a retinoid-free medium with its precursor, retinol, prompting the cells to produce RA endogenously. Our results showed that the spatiotemporal RA and FGF signaling can be manipulated with different doses of retinol, influencing gastruloid elongation and lineage specification. Gastruloids cultured in high doses of retinol showed an enrichment of ectoderm, whereas low doses resulted in early mesoderm specification and increased FGF signaling. We further used our approach to confirm that RA signaling inhibits FGF in gastruloids. Thus, retinol can modulate cellular composition in gastruloids, allowing us to investigate fate commitment of ectoderm and mesoderm progenitors in vitro.

Multiple myeloma (MM) is characterized by the production and secretion of large quantities of immunoglobulins, making this malignancy highly dependent on mechanisms that maintain cellular proteostasis. While significant clinical progress has been made by targeting the degradative branch of proteostasis, much less attention has been given to the biosynthetic branch. In this study, we demonstrated that inhibiting COPII-dependent endoplasmic reticulum (ER) export induces cell death in several MM cell lines and primary patient-derived cells. The induction of cell death was dependent on the secretory status of MM cells. Blocking ER export in secretory MM cells caused the accumulation of misfolded proteins, which activated ER-associated degradation (ERAD). Consequently, we observed an ERAD-dependent increase in the levels of free cytosolic amino acids and a subsequent activation of mTORC1 signaling. Simultaneously, we observed mitochondrial dysfunction. These alterations resulted in a mismatch between the increased energy demand due to mTORC1 activation, and the disrupted energy supply from mitochondrial impairment. This energetic imbalance results in homeostatic collapse and cell death of secretory MM cells. The therapeutic potential of the concept was demonstrated in two in vivo myeloma models. These findings suggest that the ER export machinery could be a promising therapeutic target in multiple myeloma.

Soil salinization severely restricts the growth and productivity of ginger (Zingiber officinale Rosc.). The PYL gene family encodes core components of the abscisic acid (ABA) signaling pathway, which plays a pivotal role in plant responses to abiotic stresses. However, the PYL gene family in ginger has not been systematically characterized to date. In this study, 11 ZoPYL genes were identified from the ginger genome and phylogenetically clustered into four subfamilies. Promoter sequence analysis revealed the presence of multiple stress-responsive cis-elements, and transcriptional regulatory network prediction suggested that ZoPYL proteins are potentially regulated by MYB, bZIP, and other transcription factor families. Expression profiling indicated that most ZoPYL genes were upregulated in response to ABA treatment, while salt stress induced tissue-specific expression patterns of ZoPYLs, with ZoPYL11 being significantly upregulated in both leaves and roots. Silencing of ZoPYL11 enhanced ginger sensitivity to salt stress, accompanied by reduced activities of antioxidant enzymes and increased accumulation of reactive oxygen species (ROS). Concomitantly, the expression levels of ZoPP2C and ZoSnRK2 genes were dysregulated in ZoPYL11-silenced plants, indicating disrupted ABA signaling. This study provides a comprehensive characterization of the PYL gene family in ginger and demonstrates that ZoPYL11 plays a crucial role in enhancing salt tolerance by mediating ABA signaling and antioxidant defense.

Preeclampsia is a major cause of maternal and perinatal morbidity, yet the pathogenic role of clinical metabolic marker homocysteine (Hcy) remains poorly defined. This study aimed to verify whether gestational Hcy exposure triggers preeclampsia-like phenotypes, clarify underlying decidualization-related mechanisms, and investigate the rescue effect of folate supplementation.

Acquired resistance to sorafenib remains a major obstacle to effectively treating advanced hepatocellular carcinoma (HCC). Ferroptosis has emerged as a key mechanism influencing therapeutic response. EGF-containing fibulin extracellular matrix protein 1 (EFEMP1) contributes to sorafenib resistance in HCC cells by regulating ferroptosis, but its upstream regulatory mechanisms remain elusive.

Claudin-14 (CLDN14), a tight junction protein, contributes to cell proliferation and chemoresistance in human colorectal cancer (CRC)-derived DLD-1 cells. However, small molecules targeting CLDN14 remain unexplored. Here, we identified mupirocin (MUP), a clinically approved topical antibiotic, as a modulator of CLDN14 protein expression. Quartz crystal microbalance analysis revealed interaction between MUP and recombinant CLDN14 protein with a dissociation constant (Kd) of 2.59 ± 0.54 μM. MUP did not alter CLDN14 mRNA levels but reduced CLDN14 protein stability. Pharmacological inhibition of clathrin-mediated endocytosis and lysosomal degradation significantly reversed the MUP-induced reduction of CLDN14 protein. These results suggest MUP accelerates endocytosis-lysosomal degradation of CLDN14 protein. Other antibiotics failed to decrease CLDN14 expression. Functionally, MUP increased paracellular permeability to mineral ions and enhanced the transepithelial flux of doxorubicin (DXR), an anthracycline anticancer drug, and lucifer yellow, an aqueous small compound. In DLD-1 spheroids, MUP reduced intracellular oxidative stress and nuclear factor erythroid 2-related factor 2 (Nrf2) expression. Consequently, MUP promoted intracellular accumulation of DXR and significantly potentiated its cytotoxic effects in spheroids. Moreover, MUP enhanced the antitumor efficacy of other chemotherapeutic agent oxaliplatin. Sulforaphane, an Nrf2 activator, attenuated the MUP-induced enhancement of anticancer efficacy. These findings suggest that MUP enhances anticancer drug sensitivity in CRC through lysosome-dependent downregulation of CLDN14 protein and suppression of oxidative stress signaling.

Palmatine, a protoberberine alkaloid with recognized anti-inflammatory and anti-resorptive activities, has recently attracted interest as a potential modulator of bone homeostasis; however, its direct osteo-inductive effects on human adipose-derived mesenchymal stem cells (hADMSCs) remain undefined. Given the accessibility, proliferative efficiency, and clinical relevance of hADMSCs, identifying natural small molecules capable of enhancing their osteogenic differentiation is of considerable therapeutic significance. This study investigated the osteogenic potential of Palmatine using a multi-level in vitro approach. Cytotoxicity assays established a non-toxic working concentration. Osteogenic differentiation was assessed by quantifying alkaline phosphatase (ALP) activity, calcium deposition, and matrix mineralization using Alizarin Red and von Kossa staining. Mechanistic insight was gained by evaluating antioxidant enzyme activity (superoxide dismutase, catalase) and profiling key osteogenic genes via real-time RT-PCR. Low-dose Palmatine (1 μM) enhanced hADMSC viability, whereas higher concentrations displayed dose-dependent cytotoxicity. Functional assays demonstrated that Palmatine significantly increased ALP activity and calcium content at both early (Day 7) and late (Day 14) stages. Transcriptionally, Palmatine promoted early upregulation of Runt-related transcription factor 2, osteocalcin, and osteonectin, followed by robust induction of ALP and collagen type I at later stages, reflecting canonical osteogenic progression. Furthermore, Palmatine augmented antioxidant capacity through early elevation of superoxide dismutase and later activation of catalase, suggesting a redox-associated mechanism that stabilizes osteogenic signaling and supports matrix maturation. Overall, these findings provide the first comprehensive evidence that Palmatine functions as a potent, multi-level osteo-inductive molecule in hADMSCs, highlighting its promise as a natural candidate for redox-modulated bone regenerative strategies.

African trypanosomiasis is a neglected disease affecting humans and animals in sub-Saharan Africa, caused by extracellular parasites of the Trypanosoma brucei species complex. Despite recent progress in disease control, current treatments are limited by toxicity, emerging resistance, and incomplete understanding of their mechanisms of action, highlighting the need for new targeted therapeutic strategies. In Trypanosoma brucei, post-translational modifications such as SUMOylation play essential roles in cellular processes, including cell cycle progression and variable surface antigen expression, making this pathway an attractive therapeutic target. We evaluated the effect of ML-792, a selective inhibitor of the SUMO-activating enzyme (E1, SAE), on bloodstream form parasites. Structural analysis showed conservation of key features of the E1 enzyme, consistent with susceptibility to inhibition. ML-792 treatment reduced global SUMO conjugation and nuclear SUMO levels and impaired parasite proliferation in a dose-dependent manner, with an IC50 in the micromolar range, and induced defects in cell cycle progression, including abnormal nuclei/kinetoplast configurations. These results demonstrate that pharmacological inhibition of SUMOylation disrupts essential processes in T. brucei and support the SUMO pathway as a potential target for therapeutic intervention.

Gestational exposure to environmental toxicants has been reported to be associated with epigenetic alterations in the male germ line. We have previously demonstrated that gestational arsenic exposure induces characteristic DNA methylation alterations in F1 sperm, namely the global DNA hypomethylation and the retrotransposon-associated alterations, including an increased frequency of differentially methylated cytosines (DMCs) and a predominance of hypomethylated DMCs (hypoDMCs) within long terminal repeats (LTRs) and long interspersed nuclear elements (LINEs). Hypomethylation of retrotransposon promoters is associated with increased detrimental retrotransposition and may contribute to disease development. Determining when and how these methylation alterations occur during spermatogenesis is important for elucidating the underlying molecular mechanisms.

Modulation of splicing is an established therapeutic strategy with clinical applications and potential to target specific exons to influence gene expression. Small-molecule splicing modifiers such as Risdiplam and Branaplam induce inclusion of exons typically skipped due to weak 5' splice sites. Risdiplam preferentially induces exons with an N-3G-2A-1 sequence at the 3' exon end, whereas Branaplam favors A-3G-2A-1-ending exons. However, determinants of specificity remain unclear, as many motif-matching exons are not induced. Here, we investigate the molecular basis of splicing-modulator specificity. Using biochemical assays, transcriptome analyses, and genetic perturbations, we identify sequence-dependent features that determine exon responsiveness to splicing-modulator induction. We further demonstrate that specificity can be reprogrammed through manipulation of U1 snRNA. These findings refine the determinants of splicing-modulator target space and may support identification of additional target exons and compounds.

Insulin-like growth factor 1 (IGF1) is a key somatotropic hormone that controls growth and metabolism, thereby conveying growth status to the reproductive endocrine system. The regulation of gonadotropin-releasing hormone (GnRH) to synthesize and release both the gonadotropin hormones (FSH and LH) is well documented. Numerous attempts have been made to understand the interaction of gonadotropic (reproductive) and somatotropic (growth) axis, but the dynamics of their interaction are still not clear at the pituitary level among teleost. The present study bioinformatically demonstrated protein-protein interaction of IGF1 with the proteins of both somatotropic and gonadotropic axes suggesting a cross-talk between them. In vitro, the effects of IGF1 alone or in combination with GnRH on pituitary transcripts for follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were examined. Experiments were conducted with dispersed pituitary cells of the catfish, Heteropneustes fossilis, to determine the time- and dose-dependent response of IGF1 and GnRH alone or in combination to assess fshβ and lhβ. The results showed a time-dependent effect with maximum transcripts level at 1 h of exposure. IGF1 and GnRH showed dose-dependent effects on differential expression of both transcripts when administered alone, with maximum fshβ and minimal lhβ levels at the higher dose. A synergistic impact on both the transcripts was observed in the presence of IGF1 and GnRH. Collectively, the study suggests that IGF1 plays a crucial role in regulating the synthesis of hormones in the catfish by interacting between the gonadotrophs and somatotrophs at the pituitary level.

The antitumor effects of Lasiokaurin (LAS), a natural compound derived from traditional Chinese medicine extracts, remain poorly understood in hepatocellular carcinoma (HCC). Therefore, this study investigates the anticancer effects of LAS in liver cancer. The experiment employed Wound healing assay, colony formation assays, Transwell assays, flow cytometry, Western blot analysis, and reverse transcription quantitative polymerase chain reaction (RT-qPCR) to evaluate the relationship between LAS and apoptosis, migration, proliferation, and cell cycle arrest. RNA-sequencing, RT-qPCR, and Western blot techniques were used to detect markers associated with the Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) pathway. The anti-tumor efficacy was assessed using a subcutaneous tumor-bearing mouse model. LAS promotes apoptosis and cell cycle arrest, significantly inhibiting tumor growth. These findings indicate that LAS mediates apoptosis through the JAK2/STAT3 pathway, suggesting its potential as a novel anticancer agent.

Abscisic acid (ABA) HYPERSENSITIVE GERMINATION 1 (AHG1) controls seed dormancy, germination, and early seedling establishment in response to ABA in Arabidopsis thaliana. However, whether and how AHG1 is degraded remain unclear. Here, we identified an unreported REALLY INTERESTING NEW GENE E3 ligase ARABIDOPSIS TÓXICOS EN LEVADURA 17 (ATL17) by RNA-seq analysis and investigated its functions in ABA responses using multiple biochemical, molecular and genetic methods. ATL17 positively regulates ABA-mediated suppression of seed germination and postgerminative seedling growth. It interacts with AHG1 and promotes AHG1 degradation by the 26S proteasome in the cytoplasm in response to ABA. AUXIN RESPONSE FACTOR 2 (ARF2), a crucial regulator of auxin signaling, binds to the ATL17 promoter to repress its transcription and functions in ABA responses. ATL17 prominently affects the phosphorylation of class III SUCROSE NONFERMENTING 1-RELATED KINASE 2.3 and the accumulation of ABA-INSENSITIVE 5 during ABA-arrested seed germination and early seedling growth. Our findings reveal ATL17 as a new and essential regulator of AHG1 turnover and uncover a new function of ARF2 in ABA signaling, highlighting the key roles of ATL17-regulated AHG1 degradation in governing ABA responses and likely mediating the crosstalk among ABA and auxin signaling in A. thaliana.