Salt stress is a major abiotic factor limiting plant growth and productivity. One of the primary consequences of salinity is the enhanced production of reactive oxygen species (ROS). This study investigates the role of glycolate oxidase (GOX), a key enzyme in photorespiration and a source of ROS, in the salinity response of Arabidopsis thaliana. We used two GOX T-DNA insertion mutants, gox1 and gox2, alongside wild-type (WT) plants, grown hydroponically under control conditions or exposed to 100 mM NaCl for 24 h. Results showed that shoot and root fresh weight did not differ significantly between genotypes and after 24 h of NaCl treatment. In addition, both mutants, particularly gox2, accumulated less Na+ and Cl- in shoots and roots than WT. This result was supported by ion flux analysis in roots. This fact was associated with the upregulation of key ion transporters: NHX1 (Na+compartmentalization), SOS1 (Na+ exclusion), and KUP11 and HAK5 (K+ uptake). Additionally, gox2 showed differential regulation of nitrate/Cl- transporters, with downregulation of NPF2.4, SLAH1, and SLAH3 and upregulation of NPF2.5 and NPF7.2. Furthermore, gox2 exhibited reduced oxidative damage and increased peroxidase activity under salt stress. These findings suggest that GOX2 expression may regulate plant resilience to salinity by improving ion homeostasis and antioxidative responses.

Nanobiotechnology offers sustainable strategies to enhance plant resistance by activating innate immune responses. This study evaluates the effect of chitosan nanoparticles (CNPs) on transcriptional activation of defense-associated genes in Arabidopsis thaliana.

Celastrus paniculatus (CP) is a traditional medicinal plant widely used in Ayurveda and Southeast Asian medicine for enhancing memory and treating cognitive dysfunction. Although CP has been reported to exhibit antioxidant, anti-inflammatory, and neuroprotective effects, its direct impact on activity-dependent synaptic plasticity remains insufficiently characterized. This study is aimed at investigating the effects of CP seed extract on memory performance and synaptic plasticity in a rat model, with a particular focus on AMPA receptor modulation and associated synaptic proteins. Five-week-old male Sprague-Dawley rats were randomly assigned to five groups: control, CP (80 mg/kg), donepezil (1.5 mg/kg), scopolamine (1 mg/kg), and scopolamine followed by CP. Treatments were administered daily for 14 days. Spatial memory performance was assessed using the Morris water maze. Following behavioral testing, hippocampal tissue was collected for immunohistochemical analysis of Arc protein and Western blotting of pSer831-GluA1, Arc, and PSD-95. CP-treated rats exhibited significantly reduced escape latency and increased time in the target quadrant, with outcomes comparable to those of donepezil-treated rats. In scopolamine-pretreated rats, CP administration reversed memory deficits by enhancing platform crossings and reducing escape latency. Molecular analysis revealed that CP significantly upregulated hippocampal expression of pSer831-GluA1, Arc, and PSD-95, indicating enhanced AMPA receptor trafficking and synaptic integrity. CP seed extract enhances spatial memory and synaptic plasticity by modulating critical molecular components of the glutamatergic synapse. These findings suggest that CP may support memory performance in both baseline conditions and in animals with scopolamine-induced deficits.

The link between glutaminolysis and osteoarthritis (OA) has only recently begun to be elucidated. Here, we report the association of obesity- and injury-induced cartilage damage with impaired glutaminolysis in chondrocytes. Defective glutaminolysis triggered the onset and progression of OA, with enhanced catabolism and decreased anabolism. Supplementation of α-ketoglutarate (αKG), a key component in glutaminolysis and an epigenetic factor, effectively protected cartilage against degradation in vivo via a TCA cycle- and HIF-1α-independent manner. Mechanistically, OA pathogenic factors increased H3K27me3 deposition on promoters of key glutaminolysis genes, including Slc1a5 and Gls1, leading to impaired glutaminolysis. Conversely, αKG facilitated Kdm6b-dependent H3K27me3 demethylation of not only glutaminolysis genes to rescue Gln metabolism but also Ube2o to reverse OA. Elevated Ube2o expression led to TRAF6 ubiquitination and subsequent inhibition of NF-κB signaling, thereby reversing the pathological reprogramming of glycolysis and oxidative phosphorylation and protecting against cartilage destruction. Collectively, these results demonstrated that OA pathogenic factors impair glutaminolysis through epigenetic regulation, which further exacerbate OA. Moreover, αKG restores metabolic homeostasis and alleviates OA through H3K27me3 demethylation.

Small cell lung cancer (SCLC) is known for its rapid growth and early metastasis, and SCLC patients are highly susceptible to chemoresistance. Studies have shown that the combination of ferroptosis induction and TRX pathway inhibition can significantly inhibit SCLC tumor growth, but the molecular mechanisms underlying ferroptosis in SCLC are poorly understood. In this study, we explored the regulatory role of the ALDH1L2-related metabolic pathway in SCLC chemoresistance by machine learning. We found that ALDH1L2 expression is a poor prognostic factor for SCLC patients and that high ALDH1L2 expression can negatively regulate the level of cellular lipid peroxidation and inhibit ferroptosis, thereby promoting SCLC chemoresistance. Mechanistically, ALDH1L2 interacts with the TRX2-PRDX3 antioxidant network to reduce the levels of hyperoxidized PRDX3 and oxidized PRDX3 dimers in the plasma membrane under cisplatin-induced stress and decrease cellular susceptibility to ferroptosis, thus promoting SCLC chemoresistance. In addition, we found that thiostrepton, a PRDX3 inhibitor, can synergize with chemotherapy to suppress tumor growth in SCLC, suggesting that thiostrepton might be a promising new tool for overcoming SCLC chemoresistance.

The expression patterns and potential regulatory correlates of CLDN3 in cancers remain insufficiently characterised, necessitating further investigation. We employed R software alongside bioinformatics platforms to analyse the aberrant expression of CLDN3. Experiments in vitro, including proliferation, wound healing, cell cycle progression and apoptosis assays, were conducted to evaluate the role of CLDN3 in CRC. Co-immunoprecipitation (CO-IP) and immunofluorescence analyses were conducted to investigate the interaction between CLDN3 and TRIM28. Western blotting was employed to evaluate the effect of TRIM28 on CLDN3 SUMOylation and protein stability. CLDN3 was found to be overexpressed in several cancers. Genomic alterations and promoter hypomethylation were identified as key contributors to CLDN3 dysregulation. Bioinformatic analysis suggests that CLDN3 is associated with tumour progression and poor prognosis by influencing pathways, it also contributes to immune dysregulation and chemo-resistance mechanisms. Knockdown of CLDN3 in CRC cells decreased proliferation and migration. CLDN3 overexpression was shown to reduce the sensitivity to 5-FU in CRC cells. CO-IP and immunofluorescence confirmed a direct interaction between CLDN3 and TRIM28. Western blot analysis demonstrated that TRIM28 mediates CLDN3 SUMOylation and degradation. CLDN3 influences the growth and chemotherapy resistance of CRC cells, its interaction with TRIM28 makes the TRIM28/CLDN3 axis as a promising therapeutic target for CRC.

Aberrant DNA methylation patterns are increasingly recognized as contributors to a wide range of conditions, including metabolic, oncogenic, and neurodevelopmental disorders. Nutritional factors, such as choline, can shape methylation potential via methyl group donation. The purpose of this narrative review is to synthesize current evidence on the DNA methylation landscapes underlying health and disease paradigms, with a focus on the role of choline as a compelling target for modulating epigenetic states. A comprehensive literature review search was conducted in PubMed to identify relevant studies, with additional articles retrieved from review papers.

Lineage plasticity has emerged as an important mechanism of treatment resistance in prostate cancer, increasingly associated with loss of androgen receptor (AR) signaling, and in many cases induction of stemness phenotypes and neuroendocrine features. However, targeted therapies for this stage of the disease are currently lacking. In this study, we demonstrated the critical role of the epigenetic regulator UHRF1 in the enzalutamide resistance development of prostate cancer. We have shown that UHRF1 is highly expressed in enzalutamide-resistant prostate cancer cells and its expression correlates with the loss of AR-dependent glandular features. Knocking down UHRF1 led to increased AR expression and enhanced the activity of canonical AR signaling pathway in prostate cancer cells. The combination of UHRF1 knockdown with enzalutamide treatment demonstrated synergistic tumor inhibitory effects both in vitro and in vivo. Mechanistically, UHRF1 was found to bind to AR and promote its ubiquitination and degradation. Furthermore, inhibition of UHRF1 restored AR pathway activity and re-sensitized resistant prostate cancer cells to enzalutamide. Therefore, our findings elucidate an intracellular molecular mechanism that promotes prostate cancer lineage plasticity and suggest that UHRF1 may serve as a potential therapeutic target for overcoming resistance to AR-targeted therapies.

Silicon dioxide nanoparticles (SiO2 NPs) have been shown to alleviate cadmium (Cd) toxicity in plants, but the mechanisms regarding SiO2 NPs-mediated Cd uptake and detoxification on hyperaccumulators are unknown. In this study, physiological and transcriptomic analyses were performed to investigate the impact of SiO2 NPs on the growth and Cd accumulation of the Cd-hyperaccumulator Sedum alfredii. The results showed that SiO2 NPs significantly increased root fresh weight (32.71% to 121.50%) and shoot fresh weight (22.72% to 86.36%). Simultaneously, SiO2 NPs enhanced the Cd uptake and accumulation by S. alfredii (with maximum increases of 232.14%). In the leaves, the Cd content in the phloem of the Si treatment group increased by 1.24 to 1.79-fold, indicating that SiO2 NPs enhanced the transport of Cd to the shoots. Furthermore, SiO2 NPs improved the photosynthetic parameters, with the net photosynthetic rate, transpiration rate and stomatal conductance increasing by 2.86-fold, 1.14-fold, and 1.84-fold of the control, respectively. Furthermore, SiO2 NPs significantly strengthened the antioxidative enzymes activities (SOD, CAT, POD). Transcriptomic analysis revealed that SiO2 NPs enhanced the Cd tolerance and accumulation capacity of S. alfredii by upregulating the expression of genes encoding photosynthesis-related functions (PSI, PSII), antioxidant enzymes, cell wall synthesis functions, heavy metal-responsive transcription factors (WRKY, NAC, bZIP, MYB), and Cd chelation and transport protein functions. Combining physiological and transcriptomic analyses, our findings provide the mechanisms of SiO2 NPs in promoting Cd accumulation and enhancing Cd tolerance in S. alfredii, highlighted the potential of SiO2 NPs to enhance phytoremediation efficiency.

Cytochrome P450s (CYPs) 2C subfamily (eg, 2C8, 2C9, and 2C19) and phase II enzymes such as uridine 5'-diphospho-glucuronosyltransferases (UGTs) are increasingly relevant in drug development and key targets for enzymatic induction. However, for these enzymes, weak induction signals in standard in vitro tools, such as sandwich-cultured human hepatocytes, challenge drug-drug interaction (DDI) risk assessment. This study evaluated an all-human hepatocyte coculture system (TruVivo) as a more sensitive model for CYP2Cs and UGT1A1 induction. After treatment of cells with rifampicin, carbamazepine, and phenytoin, we demonstrated robust mRNA and activity-fold-induction exceeding or meeting the 2-fold threshold in the coculture system, allowing for estimation of CYP2Cs and UGT1A1 induction parameters (IndC50, Indmax), unlike sandwich culture. Using TruVivo IndC50, Indmax of these precipitants in physiologically based pharmacokinetic (PBPK) modeling resulted in high predictive accuracy. In rifampicin studies, using TruVivo mRNA-derived data from the most sensitive donor 1 and average parameters across donors, was essential to properly predict in vivo DDI, particularly for object drugs mainly metabolized by CYP2Cs and UGT1A1, or with moderate to low CYP3A4 contribution (fm ≤ 0.5) in multipathway metabolism. For object drugs metabolized by CYP3A4 beyond 2Cs and UGTs, carbamazepine and phenytoin PBPK predictions highlighted the applicability of TruVivo uncalibrated CYP3A4 data for accurate assessment, whereas parameters calibrated against rifampicin showed a conservative trend in estimating DDI. Overall, the all-human coculture system, paired with PBPK, offers a breakthrough for CYP2Cs and UGT1A1 preclinical DDI induction risk assessment. SIGNIFICANCE STATEMENT: Cryopreserved human hepatocytes in sandwich culture show limited sensitivity toward cytochrome P450s 2C and uridine 5' -diphospho-glucuronosyltransferases induction, challenging in vitro-in vivo translation of the drug-drug interaction risk. This study confirms that TruVivo is a more sensitive in vitro model. By using physiologically based pharmacokinetic modeling, we investigated the impact of the measured induction parameters on predictive accuracy, showing TruVivo as a useful tool for cytochrome P450s 2C and uridine 5' -diphospho-glucuronosyltransferases risk assessment.

A number of published studies suggest that HIV infection accelerates epigenetic ageing. The main aim of this study was to ascertain if HIV infection is an independent factor leading to DNA hypomethylation and accelerating epigenetic ageing in men successfully treated with integrase inhibitor (INSTI)-based combined antiretroviral therapy (cART). Forty-eight (48) men living with HIV receiving INSTI-based cART and fifty (50) uninfected men in the control group were included. All participants filled out a questionnaire probing into lifestyle factors. Global and site-specific DNA methylation and expression of methyltransferase genes were examined in all participants. As well, all patients underwent basic laboratory blood tests. The results were analysed using statistical and machine learning methods. We found a strong association between HIV infection and global DNA hypomethylation as well as significant association with higher expression of the methyltransferase gene DNMT1. However, there was no association with DNA methylation levels of CNOT2, DPP6, FOXG1 and NPTX2 genes or expression levels of DNMT3a and DNMT3b. The results confirm that in men successfully treated with INSTI-based cART, HIV infection is an independent factor causing global DNA hypomethylation and increased DNMT1 expression and thus accelerating epigenetic ageing.

TP53 is a tumor-suppressor gene involved in regulating apoptosis, DNA repair, and genomic stability. Mutations in TP53 are implicated in approximately half of all detected cancers, including breast, lung, colorectal, and ovarian cancers, making it a significant target for therapeutic interventions. Many pharmaceutical drugs aim to restore TP53 function, and there is a need for predictive tools to assess how compounds may affect TP53 expression. In this study, we propose a new ensemble machine-learning model to predict the direction of TP53 relative gene expression in response to pharmaceutical compounds. Our model utilizes molecular fingerprints, descriptors, and scaffold-based features extracted from SMILES representations of compounds concatenated into a single feature vector. Trained using our newly generated benchmark dataset based on the Connectivity Map (CMap) database and addressing class imbalance with the Synthetic Minority Over-sampling Technique (SMOTE), our model achieves 62.9%, 93.9%, 40.3%, and 0.39 in terms of accuracy, sensitivity, specificity, and Matthews Correlation Coefficient (MCC), respectively. As the first-of-its-kind TP53 gene regulation prediction, our study serves as a convincing proof-of-concept that paves the way for future investigation. GenReP as a stand-alone predictor, its source code, and our newly generated benchmark dataset are publicly available.

Adult glioblastoma multiforme (GBM) is the most common primary malignant brain tumour caused by multiple molecular factors. N6-methyl-adenosine (m6A) is an abundant RNA modification that governs cellular RNA metabolism. We hypothesise that changes in m6A-modified RNA and regulatory machinery such as the writer proteins, Methyltransferase 3 (METTL3) and WT1-associating protein (WTAP), the demethyltransferase protein, and Alpha-ketoglutarate dependent dioxygenase (FTO), are driving factors of GBM development and treatment resistance. Here, we investigated m6A-RNA spatial and quantitative abundance and expression of m6A effector proteins directly in GBM tissue and patient-derived low-passage primary adult GBM and low-grade glioma (LGG) cells, and explored the consequences of m6A-RNA disruption on GBM invasive capabilities, self-renewal and responsiveness to temozolomide (TMZ). We observed that METTL3, WTAP and FTO transcript and protein expression were significantly increased in cells derived from invasive regions of GBM tumours, and elevated WTAP and FTO expression significantly correlated with poor GBM patient survival. We further found that the abundance of m6A-modified RNA in GBM tumours was significant higher in rim and invasive tissue, as well as significantly higher in patient-derived cells from GBM tumour invasive regions. Functional depletion of these effector proteins significantly altered m6A levels on and the expression of the pluripotency stem cell marker SOX2 while also impairing self-renewal and cell invasion behaviour and increasing sensitivity to TMZ. The targeting of RNA modification regulatory mechanisms reveals novel therapeutic strategies aimed at improving clinical outcomes for GBM patients.

Because prostate cancer proliferates in an androgen-dependent manner, various inhibitors of androgen production and antagonists of the androgen receptor (AR) are used as therapeutic agents. However, the emergence of castration-resistant prostate cancer has prompted the development of additional treatment strategies. In this study, we focused on the antiprostate cancer effects of vitamin D3 and examined novel antiproliferative effects through the crosstalk with androgen signaling. In human prostate cancer LNCaP cells, homeobox C9 (HOXC9) was identified as a common regulated target gene by dihydroxytestosterone and 1α,25-dihydroxyvitamin D3, but in opposite directions. Ligand-stimulated AR and vitamin D receptor competitively shared binding sites in the HOXC9 regulatory region, but dihydroxytestosterone stimulation preferentially suppressed HOXC9 expression due to the stronger binding properties of AR and the induction of DNA methylation. Forced expression of HOXC9 inhibited androgen signaling to eliminate the androgen-dependent proliferation by associating with the AR transcription complex, in part due to interference with AR binding to some of its targets in LNCaP cells. In summary, this study provides evidence for the involvement of HOXC9 in antiproliferative effects through a regulatory mechanism mediated by a crosstalk between vitamin D receptor and AR.

Lavender essential oil (LEO) is commonly used in aromatherapy for stress reduction, relaxation and recovery from (muscle) fatigue. However, molecular mechanisms underlying its potential physiological effects on the skeletal muscle remain unclear. This study investigates whether LEO affects the intracellular signaling pathways in skeletal muscle cells that respond to physical activity. Prior to the experiment, GC-MS analysis confirmed linalyl acetate and linalool as the main components of LEO used in this study. Transdermal permeability was assessed using a reconstructed human epidermis model, which showed that linalool permeated the epidermal layer, while linalyl acetate showed minimal permeation. Following this confirmation, the differentiated C2C12 myotubes were treated with LEO in an in vitro muscle contraction model using electrical pulse stimulation (EPS). LEO significantly increased Interleukin 6 (IL-6) mRNA expression under EPS, and DNA whole-genome microarray analysis showed that LEO induced different gene expression profiles depending on the contraction state of the muscle cells. These results provide the first molecular evidence that LEO modulates skeletal muscle gene networks in a stimulation-dependent manner and may indicate its potential use as an aid to recovery (from fatigue) after exercise. Notably, the skin permeation of LEO components showed a saturation trend at concentrations above 5%, suggesting the presence of an optimal concentration range for topical application in sports aromatherapy.

DNA methylation is a conserved regulatory mechanism of gene expression, genome stability, and development, and is highly associated with the effective induction of defense responses for plant priming. In the Green Deal era, the use of plant defense inducers (PDIs), compounds that activate defense and prime plants against imminent pathogen attacks, is a safe and environmentally sustainable approach to support plants against pathogens. Though efforts have succeeded at deciphering part of the mode of action of PDIs, more information is needed to understand the underlying pathways of their effectiveness. Here, salicylic acid (SA), loaded in chitosan nanoparticles, increased hypomethylation by more than 25% for 56 genomic regions that corresponded to defense-related genes, such as pectin lyases, defensins and leucine-rich repeat transmembrane protein kinases against the biotrophic fungal pathogen Podosphaera xanthii. A genomic region of the promoter of SKP1A, which is a core member of the SCF E3 ubiquitin ligase complex, was found to be a differentially methylated region (DMR), with 60% hypomethylation, both after PDI application and pathogen inoculation, possibly indicating a similar activation mechanism. Examination of this DMR revealed the presence of SA-, auxin-, and defense-related cis-elements. Investigation of the proteins associated with the above cis-elements showed significant upregulation in expression after PDI. Moreover, association of the identified DMR with transcriptomics showed enrichment of the SA pathway. Overall, these findings shed light on the epigenetic mechanisms that underlie SA-related defense priming in plants.

Thiocyanate (SCN-), a persistent inorganic contaminant widely present in industrial wastewater, poses severe risks to plant growth and photosynthesis. Hydrogen sulfide (H2S) is an emerging gaseous signaling molecule involved in the regulation of plant stress responses; however, its role in modulating Rubisco energy metabolism and activation under SCN- stress remains unclear. Here, we investigated the effects of exogenous H2S on magnesium homeostasis, ATP/NADPH metabolism, Rubisco activation, and photosynthetic performance in rice seedlings exposed to SCN- stress via physiological, biochemical, and transcriptional approaches. We found that exogenous H2S significantly increased Mg2+ accumulation, enhanced H+-ATPase and Mg2+-ATPase activities, and promoted Rubisco activase (RCA) abundance and activity. These changes were accompanied by reduced steady-state ATP and NADPH contents, indicating that increased energy consumption was driven by accelerated Calvin cycle turnover. At the transcriptional level, H2S regulated key genes involved in ATP hydrolysis, Mg2+ transport, Rubisco activation, and chlorophyll biosynthesis. Consequently, the chlorophyll content, stomatal conductance, and transpiration rate improved under SCN- stress. Collectively, our results demonstrate that exogenous H2S enhances photosynthetic efficiency and Rubisco carboxylation capacity by coordinating Rubisco energy metabolism and activation.

The incidence and mortality rates of lung adenocarcinoma (LUAD) continue to rise, highlighting an urgent need for novel therapeutic targets. In this study, bioinformatics analysis revealed that members of the metabotropic glutamate receptor (mGluR) family are significantly correlated with the expression profile, prognosis, genetic mutations, and tumor immune microenvironment of LUAD, with GRM5 being the most significantly associated member. Overexpression of GRM5 has been shown to inhibit LUAD proliferation and induce apoptosis, while cinchonine (CN) treatment further enhances these effects, suggesting that CN may act as a GRM5 agonist to synergistically exert antitumor activity. Transcriptome sequencing further identified four key downstream targets and their associated signaling pathways. In summary, this study confirms that GRM5 can serve as a potential prognostic biomarker and therapeutic target for LUAD, while the small-molecule compound CN shows promise as an antitumor candidate drug targeting GRM5.

Metastasis accounts for most cancer-related deaths and hepatocellular carcinoma (HCC) is no exception. Midkine (MDK) is a multifunctional secreted protein elevated in HCC with a vague role in HCC. In this study, we used bioinformatics to verify MDK expression in HCC tumors, and next, we inhibited the MDK protein in invasive Hep3B cells using an MDK inhibitor (iMDK) both in vitro and in vivo. Our results showed that iMDK promoted cell migration and enhanced lamellipodia formation while at the same time downregulating the expression of cell-matrix adhesion genes. In order to also consider forces exerted by the surrounding matrix, we performed cell adhesion, transwell invasion, and 3D tumor spheroid invasion assays in two different stiffness conditions. Adhesion and invasion always exhibited opposite patterns, with adhesion being inhibited in soft matrix environments, accompanied by increased invasion, and a reverse effect in stiff environments. In vivo experiments where cells pre-treated with iMDK were implanted to zebrafish embryos showed overall reduced metastasis, verifying that MDK is a central mechanotransduction regulator that enables HCC cells to adapt their metastatic strategies to ECM stiffness. Thus, MDK inhibition effectively disrupts mechanosensitive coordination during metastasis, highlighting its potential as a therapeutic target.

With the growing global population and the challenges posed by climate change on agriculture, improving seed germination quality has become an urgent task. Nanotechnology, particularly silver nanoparticles (AgNPs), offers a promising approach to this issue. However, their long-term environmental impact and health risks require further evaluation.This review first explores the physicochemical properties of AgNPs and their effects on plant growth and seed development. Next, the review discusses the mechanisms by which AgNPs enhance seed resistance to pathogens, regulate reactive oxygen species (ROS) balance, activate key metabolic enzymes, induce metabolite accumulation, and modulate plant hormone levels. Additionally, the review explores how AgNPs influence seed gene expression, proteomic networks, and the germination microenvironment. Given the lack of field data on long-term low-dose exposure and challenges in monitoring morphological transformation, the review also evaluates the potential risks of AgNPs in agriculture. These risks include their accumulation in the food chain, environmental transformation, and long-term effects.The review aims to summarize the mechanisms by which AgNPs impact seed germination and plant growth, providing a theoretical basis for their cautious use in agricultural and horticultural practices, while considering their environmental fate and health risks.