Phenotypic, physiological and molecular analyses demonstrate that MdNAC1 enhances plant tolerance to cadmium stress. Cadmium (Cd) is a highly toxic heavy metal that severely impairs the growth and development of crops. NAC transcription factors are key regulators of plant abiotic stress responses, but their roles in apple (Malus domestica) Cd stress responses remain largely unclear. Here, we identified the apple NAC gene MdNAC1, which exhibits elevated transcriptional levels under Cd stress. The increased abundance of MdNAC1 upregulates the expression of antioxidant enzyme genes, thereby significantly enhancing antioxidant enzyme activity in transgenic plants. This in turn lowers reactive oxygen species (ROS) accumulation and alleviates oxidative damage to plant growth and development under cadmium stress. Ectopic expression of MdNAC1 in Arabidopsis reduced Cd accumulation by negatively regulating the expression of Cd uptake and transport genes, thereby enhancing Cd tolerance. Further Quantitative Real-time PCR analysis revealed that the expression of multiple Cd uptake and transport-related genes was suppressed in transgenic apple seedlings to regulate Cd stress resistance. Moreover, MdNAC1 overexpression enhanced biomass and chlorophyll content in transgenic plants under Cd stress. Our findings indicate that MdNAC1 enhances plant tolerance to Cd through multiple mechanisms in both apple and Arabidopsis, including increased antioxidant activity, ROS scavenging, increased biomass production, and regulation of Cd uptake and transport. These findings provide key candidate genes for fruit tree breeding programs and offer novel insights into the regulatory mechanisms mediating Cd stress resistance.

To evaluate the effects of rexinoid NEt-3IB on desiccating stress-induced dry eye, as well as monocyte/macrophage gene expression and cellular trajectory.

Background/Objectives:Syringa vulgaris L. (common lilac) is one of the most popular ornamental plant species. Through the ages, many parts of S. vulgaris, including fruits, flowers, leaves, and branches, have been used in folk medicine due to their beneficial biological activity. Lilac flowers are the basis of many supplements available on the market. Moreover, its petals and flowers are edible and are an aromatic ingredient in preserves and desserts. However, the data about the antioxidant properties of various parts of S. vulgaris is limited only to the in vitro antioxidant capacity of the extracts-so far, the effect of S. vulgaris flower extract on the parameters of oxidative stress in biological materials, including plasma, has not been demonstrated. Therefore, the aim of our study was to investigate the protective effects of the extract from S. vulgaris L. flowers against oxidative stress in human plasma, and its influence on the coagulation process in vitro. Methods: We measured the levels of three parameters of oxidative stress in human plasma treated with H2O2/Fe2+ (the donor of hydroxyl radicals): lipid peroxidation (based on the level of thiobarbituric acid reactive substances (TBARS)), protein carbonylation, and thiol oxidation. Ascorbic acid (vitamin C) was used as a reference antioxidant. In addition, we studied the effect of the extract on three coagulation parameters of human plasma-activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). We also compared the biological properties of the extract from S. vulgaris flowers with the properties of a phenolic extract from Taraxacum officinalis (dandelion) flowers, as they have proven antioxidant activity in both in vitro and in vivo models and can modulate hemostasis in vitro. Results: Our UHPLC-HRMS analyses of S. vulgaris extract led to a tentative identification of 50 compounds, mainly phenolics and secoiridoids. For the first time, the present study demonstrated that the extract from S. vulgaris flowers (at the concentrations of 1-50 µg/mL) significantly reduced plasma lipid peroxidation and protein carbonylation induced by H2O2/Fe2+. Moreover, the concentrations of 1-25 µg/mL significantly reduced the oxidation of thiol groups in plasma treated with H2O2/Fe2+. The anticoagulant tests also demonstrated that S. vulgaris flowers extract, at physiologically relevant concentrations (1-50 µg/mL), did not affect blood clotting times in vitro, suggesting that it is hemostatically safe. Conclusions: Despite the differences in composition, the extracts from lilac flowers and dandelion flowers exhibited similar protective effects against oxidative damage to human plasma components. However, the extract from S. vulgaris flowers had a stronger inhibitory effect on lipid peroxidation than the extract from dandelion flowers.

Cluster of Differentiation 47 (CD47) functions as a key "don't-eat-me" signal that enables cancer cells to evade macrophage-mediated immune clearance. GV1001, a 16-amino-acid peptide derived from human telomerase reverse transcriptase (hTERT), has been reported to exhibit antitumor and anti-inflammatory properties and to downregulate CD47 expression in human cells. In this study, we investigated whether GV1001 modulated CD47 expression and enhanced antitumor immunity in oral squamous cell carcinoma (OSCC). In vitro, GV1001 significantly reduced CD47 expression in both murine and human OSCC cells in dose- and time-dependent manners, resulting in a marked increase in macrophage-mediated phagocytosis. Mechanistically, GV1001 suppressed CD47 promoter activity and inhibited multiple upstream regulator expression in murine and human OSCC cell lines, while exerting minimal effects on normal human keratinocytes and fibroblasts. In vivo, GV1001 significantly inhibited tumor growth, suppressed CD47 expression, increased macrophage infiltration, and induced tumor cell necrosis and apoptosis in both murine OSCC syngeneic graft model and human OSCC xenograft model. GV1001 administered alone or in combination with cisplatin produced antitumor effects. Collectively, these findings demonstrate that GV1001 functions as a potent immunomodulatory anticancer peptide that downregulates CD47 expression and restores macrophage-mediated tumor clearance, highlighting its potential as a therapeutic strategy for OSCC.

Filamentous fungi exhibit high heavy metal resistance; elucidating their resistance mechanisms is of practical importance for fungal utilization and for engineering other microorganisms. However, the molecular basis of copper tolerance in filamentous fungi remains poorly understood, with few studies addressing this specific trait. Previously, we isolated a copper-hyper-resistant strain, P. janthinellum GXCR, and generated two mutagenized derivatives, EC-6 and UC-8. To investigate copper resistance, wild-type GXCR (WT) and mutants EC-6 and UC-8 were subjected to integrated physiological, biochemical, and transcriptomic analyses. Copper tolerance followed the rank order: WT > UC-8 > EC-6. Supplementation with Mn2+ or exogenous proline enhanced copper resistance. Under copper stress, intracellular reactive oxygen species (ROS) levels increased in all strains, correlating dynamically with activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as malondialdehyde (MDA) content, with all exhibiting a biphasic response: an initial rise followed by a decline with increasing Cu2+ concentration. WT accumulated less Cu and Cd but more Cr (at high concentration) than the mutants. In contrast, intracellular Pb accumulation in all three strains decreased monotonically with rising Pb doses. RNA-seq of WT and EC-6 grown in TYB with 0, 0.5 and 3 mM Cu2+ identified 8 copper-resistance-related genes, verified by real-time quantitative reverse transcription PCR (RT-qPCR). Weighted gene co-expression network analysis (WGCNA) clustered genes into 10 modules; integrating physiological data identified 10 traits, and the four most correlated modules yielded 116 hub genes mostly linked to energy metabolism, cell components and transporters. copA and ATP7, encoding Cu2+-exporting ATPases, were identified as central regulators of copper homeostasis and key contributors to enhance copper tolerance. These findings provide molecular insights into copper resistance of filamentous fungi and valuable genetic targets for rational strain engineering.

The prediction of drug response would significantly improve the treatment of lung cancer. Tumor heterogeneity and complex signal transduction pathways lead to varied treatment effects among patients, but traditional computational approaches struggle to model the nonlinear, high-dimensional relationship between genes and drug responses. In order to develop a Generative Adversarial Network (GAN)-based model that can predict drug-induced gene expression profiles from lung cancer cell lines, we developed GRIP-Lung (Generative Model of Response to Drug-Induced Perturbation in Lung Cancer). By making use of biologically informed embeddings of cell line identity as well as drug treatment conditions, this model is able to gain a fairly good understanding of cell types and their transcriptional perturbations induced by different drugs. The GRIP-Lung model displayed reasonably good prediction ability in terms of predictive accuracy and showed high concordance between the predicted and experimental expression profiles. We not only predicted transcriptional changes induced by drug therapy but also used single-sample Gene Set Enrichment Analysis (ssGSEA) to classify post-treatment response states based on characteristic molecular biomarkers, offering a means for selecting effective drugs to target specific heterogeneity within lung tumors. The proposed GRIP-Lung framework faithfully reproduces drug-induced transcriptional perturbations in lung cell line models. By integrating biologically informed embeddings and adversarial learning, the model advances drug response prediction. This makes it a flexible computational tool for drug repositioning.

Arsenic induces apoptosis in both cancerous and non-cancerous cells. The mechanism of arsenic-induced apoptosis is complex. We previously demonstrated that the antioxidant acetyl L-carnitine prevented sodium arsenite-induced apoptosis in zebrafish embryos. To gain more insight into the mechanism of arsenic-induced apoptosis, we explored the effect of another antioxidant, N-acetylcysteine (NAC). Co-treatment of sodium arsenite with 1 or 2 mM NAC had no effect on zebrafish development. There was a significant but partial reduction in apoptosis in the embryos co-treated with sodium arsenite and 1 mM NAC, while embryos treated with 1 mM NAC alone showed the loss of normal apoptosis that was observed in the control embryos. Complete abolition of apoptosis occurred in embryos co-treated with sodium arsenite and 2 mM NAC; however, 2 mM NAC alone resulted in 100% mortality, indicating antioxidant toxicity at high doses. NAC (1 mM) did not prevent sodium arsenite-induced increase in motor neurons, suggesting that arsenic-induced apoptosis and supernumerary motor neuron development are mediated via distinct pathways. To determine whether NAC prevented arsenic-induced apoptosis via reactive oxygen species (ROS) signaling, we assessed ROS levels and oxidative modification of proteins (carbonylation) using an OxyBlot assay. Neither sodium arsenite nor NAC altered protein oxidation, ROS levels, or p53, a pro-apoptotic protein, transcript levels. Additionally, dicoumarol, an inducer of p53 protein degradation, did not inhibit sodium arsenite-induced apoptosis. These results indicate that protein oxidation and p53 signaling are not involved in arsenic-induced apoptosis and that NAC prevents arsenic toxicity in zebrafish embryos through a hitherto unknown mechanism.

Tumor microenvironments, particularly hypoxia and inflammation, heavily influence colorectal cancer (CRC) pathogenesis by altering polyamine metabolism. Identifying natural compounds targeting these vulnerabilities remains critical. Integrating untargeted metabolomics, network pharmacology, and a human endogenous metabolite library screen, we identified apigenin (API) as a potent anti-CRC candidate. API significantly inhibited the proliferation, migration, and invasion of RKO and HCT116 cells in vitro and suppressed xenograft tumor growth in vivo. Crucially, high-throughput screening revealed that polyamines rescued CRC cells from API-induced cytotoxicity. Mechanistically, API exerts its effects by dismantling a newly identified HIF-1α/SMOX positive feedback loop. In CRC, HIF-1α transcriptionally activates spermine oxidase (SMOX), while SMOX-driven polyamine metabolism fuels the TLR4/MyD88 inflammatory cascade to continuously stabilize HIF-1α. API acts as a "circuit breaker" for this axis, significantly reducing the spermidine/spermine ratio and downregulating inflammatory signaling. Ultimately, API effectively remodels polyamine metabolism and suppresses CRC progression by disrupting the HIF-1α/SMOX and TLR4/MyD88 pathways, offering a novel metabolic mechanism for API in CRC therapy.

Liver cancer is one of the most prevalent and lethal cancers worldwide, characterized by poor prognosis and limited treatment options. Triptophenolide (TRI), a diterpenoid compound, has shown anti-proliferative activity in breast and pancreatic cancers, but its role in liver cancer remains largely unexplored. In this study, TRI significantly inhibited the proliferation of HepG2 (hepatoblastoma) and HuH7 (hepatocellular carcinoma) cells in a dose-dependent manner, with IC50 values decreasing from 279.9 to 229.4 µg/mL (24-48 h) in HepG2 and from 441.1 to 282.6 µg/mL in HuH7. Colony formation assays confirmed the suppression of HCC cell growth. TRI also promoted apoptosis, increasing apoptotic rates to 68.99% in HepG2 and 43.34% in HuH7 at 400 µg/mL (48 h). Cell cycle analysis revealed S-phase arrest, with TRI raising the S-phase population to 42.02% and 45.38%, respectively. Mechanistically, TRI upregulated pro-apoptotic genes (TP53, CASP3/9/10, BAX, BAK1, BID, BIM) and proteins, activating the mitochondrial apoptotic pathway. In vivo, TRI (10 mg/kg) markedly reduced tumor volumes in HepG2 and HuH7 xenografts compared with controls, without obvious systemic toxicity. These findings suggest that TRI exerts anti-proliferative, pro-apoptotic, and cell cycle regulatory effects in HCC. However, further preclinical studies are warranted to elucidate its mechanisms and evaluate its safety profile.

Corneal alkali burns represent one of the most severe forms of ocular surface injury and frequently result in persistent inflammation, corneal neovascularization, stromal remodeling, and permanent visual impairment. Current therapeutic approaches incompletely control the inflammatory mechanisms that sustain pathological angiogenesis and tissue disorganization. In this study, we evaluated the effects of a transglutaminase-binding fusion protein (FP) in a rat model of alkali-induced corneal injury. Following standardized alkali burns, animals were treated topically with FP, secretory leukocyte protease inhibitor (SLPI), or Buffer. Corneal epithelial healing, opacity, and neovascularization were assessed clinically and by digital image-based quantification, while histological and immunofluorescence analyses were used to evaluate stromal organization and vascular invasion. Molecular mechanisms were investigated by RT-qPCR and Western blot analysis of key inflammatory, angiogenic, and signaling mediators. FP treatment significantly accelerated corneal re-epithelialization, reduced corneal opacity, and markedly attenuated corneal neovascularization compared to SLPI and Buffer controls. These effects were associated with coordinated downregulation of pro-inflammatory cytokines and angiogenic mediators, including TNF-α, IL-17, VEGF, and cPLA2. Notably, FP suppressed transglutaminase 2 expression and induced early and sustained downregulation of NF-κB pathway components, identifying modulation of an upstream inflammatory pathway central to corneal angiogenesis and stromal remodeling. Collectively, these findings demonstrate that FP effectively limits inflammation-driven corneal neovascularization and tissue remodeling following alkali injury, supporting its potential as a disease-modifying therapeutic strategy for inflammatory ocular surface disorders.

Atherosclerosis (AS) is a cardiovascular disease characterized by diverse etiological factors and complex pathological mechanisms. In recent years, the role of long non-coding RNAs (lncRNAs) in AS has received increasing attention. Research shows that lncRNAs regulate key biological processes involved in AS, such as vascular endothelial function, proliferation and migration of vascular smooth muscle cells (VSMCs), macrophage polarization, and lipid metabolism, through various mechanisms, including epigenetic modification, transcriptional regulation, and post-transcriptional control. As important components of traditional medicine, Chinese herbal monomers and compounds have been found to modulate the expression of lncRNAs, thereby improving vascular endothelial function, reducing lipid deposition, and inhibiting inflammatory responses, ultimately exerting anti-atherosclerotic effects. This review systematically examines the role of lncRNAs in the disease mechanism of AS and summarizes recent advances in Traditional Chinese Medicine (TCM) interventions targeting lncRNA expression for the treatment of AS, offering new insights and directions for the prevention and management of AS with Chinese medicine.

Herein, we have identified the polyfunctionalized 1-(phenylsulfonyl)-1H-indole-2-carboxylic acid derivative MTP150 for the treatment of neurodegenerative diseases owing to its efficacy in reducing protein aggregation, modulating matrix metalloproteinase activity, mitigating neuroinflammation, and enhancing DNA damage repair pathways across in vivo Caenorhabditis elegans models of Alzheimer's disease, Parkinson's disease (PD), and Huntington's disease. Further experiments in an in vivo Drosophila model of PD showed that MTP150 increased motor performance, reduced oxidative stress levels, and restored mitochondrial function in model flies. In addition, MTP150 exhibited neuroprotective effects in PD model cells, thereby supporting its therapeutic potential for this disease.

Erlotinib (ER) and gefitinib (GB) are frequently prescribed for patients with advanced lung adenocarcinoma (ADC). Although both ER and GB significantly extend median survival, several patients experience early disease relapse due to treatment resistance. This study aimed to identify common genes associated with acquired resistance to ER or GB. This integrative analysis of transcriptomic data identified differentially expressed genes by comparing sensitive PC-9 cells with ER-resistant (PC-9/ER) or GB-resistant (PC-9/GB) cell lines. A Venn diagram was used to identify common genes. Candidate genes were confirmed in our generated drug-resistant cell lines. After six months, the PC-9/ER and PC-9/GB cell lines exhibited 40.7-fold and 109.2-fold increases in resistance to ER and GB, respectively. Flow cytometric analysis demonstrated that both resistant cell lines were resistant to cell cycle arrest and apoptosis induced by ER or GB. Through integrated analysis, we found that two genes, KRT13 and CEACAM5, were up-regulated in both the PC-9/ER and PC-9/GB cell lines. The expression patterns of these two genes did not align with the transcriptome, with only CEACAM5 exhibiting consistent gene and protein expressions. Our findings indicate that CEACAM5 serves as a biomarker for predicting the response of patients to ER and GB treatments.

Airway remodeling is a process that occurs in chronic obstructive diseases, such as asthma and COPD. It is associated with adverse changes in the structure and function of the airways. An increasing amount of literature points to the potential protective effects of vitamin D and caffeine against inflammation and fibrosis. The aim of the study is to evaluate the effect of calcitriol and caffeine on the expression of genes and proteins associated with airway remodeling. The Calu-3 cell line was treated with TGF-β, calcitriol, and caffeine in different combinations. Subsequently, the expression of VDR, CDH1, VIM, MMP-2, and MMP-9 were examined at the mRNA and protein levels using real-time PCR and Western blot, respectively. One-way analysis of variance was used to determine differences in several groups. Both calcitriol and caffeine were associated with a decrease in the expression of MMP-2 and VIM in TGF-β-treated cells (p = 0.01 and p = 0.006, respectively). Both compounds also reduced the expression of MMP-9 in comparison to TGF-β alone (p = 0.03), though the changes in MMP-9 protein levels did not reach statistical significance. Calcitriol was associated with a decrease in CDH1 expression at both levels in comparison to TGF-β (p < 0.0001 and p = 0.02, respectively). A potential synergistic effect was demonstrated for CDH1 at the mRNA level and for the vitamin D receptor at the protein level. Both vitamin D and caffeine may influence the pathways involved in airway remodeling. Preliminary in vitro findings suggest a potential role of these substances for future therapeutic strategies targeting obstructive diseases; however, the observations require confirmation in further in vivo studies.

Uveal melanoma (UM) represents an uncommon intraocular malignancy with high aggressiveness. Dysregulation of ferroptosis has been associated with UM progression. Dihydroartemisinin (DHA), a natural derivative of Artemisia annua, exhibits potent antitumor activity with a favorable safety profile, yet its role in ferroptosis regulation in UM remains unclear. Here, we showed that DHA significantly reduced the proliferation and invasiveness of UM cells-both primary and secondary-with effects intensifying over time and dose. Transcriptomic analysis indicated that DHA may exert antitumor effects by modulating the ferroptosis-related pathway, characterized by upregulating heme oxygenase-1 (HO-1) and downregulating the SLC7A11 (xCT)/GPX4 axis, leading to iron accumulation, increased ROS and lipid peroxidation, and mitochondrial dysfunction. Iron chelators and pancaspase inhibitors partially reverse these effects, whereas HO-1 inducers enhance them. Overall, our results suggest that DHA suppresses UM progression by inducing ferroptosis and mitochondrial dysfunction, while the HO-1 and xCT/GPX4 pathways may contribute to these effects. DHA may represent a potential therapeutic approach for UM, warranting further investigation.

The cytotoxic effect of islet amyloid polypeptide (IAPP) misfolding and aggregation has a well-recognized role in the pathogenesis of type 2 diabetes mellitus, mediated by failure of the beta cell's protein quality control system to rescue the cell from overwhelming proteotoxic stress induced by IAPP aggregates, ultimately leading to apoptosis. A small but growing body of research also links IAPP-mediated proteotoxic stress to the pathogenesis of type 1 diabetes and to the functional decline of transplanted islets. Among the most promising therapeutic approaches under investigation are small organic molecules that may act as direct chemical chaperones to prevent IAPP aggregation, promote the activity of endogenous chaperones, or alter gene networks of the unfolded protein response (UPR) to promote pro-survival rather than pro-apoptotic pathways in response to IAPP-mediated proteotoxic stress. Compounds warranting special attention include 4-phenylbutyrate (PBA), tauroursodeoxycholic acid (TUDCA), and epigallocatechin gallate (EGCG), as each has a growing body of evidence supporting their ability to ameliorate this process, and given that each of these are already known to have good safety profiles in humans, potentially accelerating the timeline to interventional studies. This review explores the evidence for IAPP-mediated proteotoxicity in multiple forms of diabetes, the mechanisms of cytotoxicity at different levels of the cell's protein quality control systems, how these small organic compounds may act on these processes including new insights on the role of thioredoxin-interacting protein (TXNIP), and the current evidence supporting each of these compounds in mitigating diabetogenesis.

Ochratoxin A (OTA), a prevalent food contaminant, is closely linked to the development of various cancers, including clear cell renal cell carcinoma (ccRCC). However, the potential mechanisms remain to be explored. In this study, we employed network toxicology, machine learning, and molecular docking techniques to systematically investigate the potential molecular mechanisms underlying OTA-associated ccRCC. We normalized transcriptional data from two Gene Expression Omnibus (GEO) datasets and analyzed it using differential expression analysis and weighted gene co-expression network analysis (WGCNA), identifying 3224 ccRCC-associated target genes. These were intersected with 232 predicted OTA target genes, yielding a total of 56 overlapping targets. The results of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that these targets were primarily enriched in critical biological processes, including extracellular matrix remodeling, immune microenvironment regulation, signaling pathway transduction, cellular metabolism, and protein homeostasis. Machine learning analysis identified "glmBoost + RF" (a sequential combination of feature selection and classifier) as the optimal model, from which nine key genes were extracted. SHapley Additive exPlanations (SHAP) analysis revealed five core genes (IGFBP3, ITGA5, PYGL, SLC22A8, LTB4R), with IGFBP3 and ITGA5 serving as the principal driver genes of the model. Validation of the model's diagnostic efficacy and single-cell transcriptome analysis indicated that the core genes exhibited significant differential expression patterns, cell-type-specific expression characteristics, and high independent diagnostic efficacy. Molecular docking analyses predicted stable interactions between OTA and the core target proteins. These findings suggest potential molecular links between OTA exposure and ccRCC, providing a foundation for hypothesis generation and future experimental validation.

Walnut (Juglans regia) is an economically significant woody oil tree species widely cultivated in China. However, its production is increasingly threatened by extreme low-temperature events, such as unseasonal frosts and late-spring cold. Salicylic acid (SA) is a key phytohormone known to enhance cold tolerance in plants, yet its underlying mechanism in walnut remains unclear. In this study, we present the first integrated analysis combining physiological measurements, transcriptomics, and metabolomics to investigate how exogenous SA improves cold tolerance in walnut leaves. Our results showed that SA treatment significantly increased the accumulation of soluble sugars, chlorophyll, and proline, enhanced peroxidase (POD) activity, and reduced malondialdehyde (MDA) levels under cold stress. Multi-omics analysis revealed that SA modulated the expression of genes involved in multiple hormone signaling pathways, including those of SA, auxin, jasmonic acid, and abscisic acid, and altered corresponding hormone levels. Notably, carbohydrate metabolism emerged as a central pathway mediating SA-induced cold adaptation. Weighted gene co-expression network analysis (WGCNA) further identified several core candidate genes, such as JrTGA, JrPP2C, JrTPS, and JrBAM, which may play key roles in this process. Collectively, this study provides the first multi-omics perspective on the regulatory network underlying SA-enhanced cold tolerance in walnut. These findings offer both a theoretical and technical foundation for applying SA in cold-resistant walnut cultivation and contribute to the development of stress-resilient production strategies.

Neonicotinoids are widely used pesticides that have caused a catastrophic decrease in bee and bumblebee populations worldwide. In addition to insects, neonicotinoids induce toxic effects in other species, including lizards, birds, and mammals. Previous studies have shown that gestational exposure to thiacloprid promotes transgenerational effects in the testes and thyroid. In this project, we described the epigenetic effects of thiacloprid on prostate tissue in directly exposed F1 and non-directly exposed F3 outbred Swiss male mice. We used paraffin sections for morphological analysis and frozen tissue for immunofluorescence analysis, RT-qPCR, and protein analysis. We purified histones and analyzed them through Western blot. We used ChIP-qPCR for histone H3K4me3 occupancy analysis. A tendency to increase in epithelial hyperplasia in F1 but not in F3 prostate was detected. Elevated levels of phosphorylated histone H3 at serine 10, a marker of mitosis, in both the F1 and F3 prostates were noted. A significant increase in the level of the Ki-67 marker of proliferation was detected in the F1 but not in the F3 anterior prostate. Hox gene expression was upregulated in the F1 and downregulated in the F3 prostate. The changes in gene expression were positively associated with histone H3K4me3 alterations at the promoters of the Hoxa and Hoxb13 genes. We determined that regions of Hox genes that play important roles in prostate development had altered DNA methylation in the sperm of F1 and F3. These alterations in DNA methylation were negatively related to gene expression. This is an observational study, as it was part of our previous research on the effects of thiacloprid on the testis and thyroid. Our analysis revealed that gestational exposure to thiacloprid induced an increase in cell proliferation in the prostates of directly exposed F1. Some persistent epigenetic alterations in the prostate of F3 males were not associated with phenotypic changes.

Rare earth elements (REE), such as gadolinium (Gd) and erbium (Er), are increasingly recognised as emerging environmental contaminants due to their widespread use in industrial processes, electronics, and medical imaging applications. Despite their extensive presence in aquatic ecosystems, little is known about their developmental toxicity. In this study, Xenopus laevis embryos were exposed to environmentally relevant concentrations of Gd and Er during critical early developmental stages. The assessed endpoints included survival, malformations, growth (body length), and heart rate. Both Gd and Er caused significant sublethal effects, including increased axial malformations, reduced growth, and altered cardiac activity. To explore potential mechanisms of toxicity, the expression patterns of key developmental genes (fgf8, bmp4, sox9, egr2, rax1, pax6) and pro-inflammatory cytokines (tnfα, il1β, p65) were analysed using Real-Time PCR. The results showed dysregulation of gene expression, indicating disruption to pathways involved in morphogenesis and neurodevelopment. Elevated reactive oxygen species levels suggested that oxidative stress was a contributing factor. Raman spectroscopy confirmed biochemical changes affecting proteins, lipids, and nucleic acids, providing evidence of cellular stress and metabolic imbalance. Overall, our findings demonstrate that even low-level exposure to Gd and Er can impair amphibian embryonic development and disturb molecular homeostasis. These results emphasise the ecological risks of REE pollution and highlight the importance of ongoing environmental monitoring and long-term toxicological research.