In clinic, once the ovulated oocytes cannot be fertilized timely, they begin to undergo post-ovulatory aging, which impacts the embryo development and offspring health. Until now, there are no better methods to delay post-ovulatory oocyte aging. Epigallocatechin gallate (EGCG) is the most abundant bioactive component of tea polyphenols that are beneficial for alleviating the deleterious influence of environmental factors on oocyte quality. In the present study, we examined the role of EGCG in delaying oocyte aging after ovulation in vitro. Ovulated oocytes were treated with EGCG at different concentrations, and the fragmentation rate, induced by aging, was significantly reduced by EGCG at 50 μM in vitro. To further elucidate the influence of EGCG on the quality of post-ovulatory aged oocytes, we examined the spindle morphology of aged oocytes. The results showed that EGCG significantly decreased the abnormal rate of spindle morphology in post-ovulatory aged oocytes. The high level of reactive oxygen species (ROS) and mitochondrial dysfunction in post-ovulatory aged oocytes were also improved by EGCG, which might be a reason for the reduced apoptosis. The reduced sperm binding capacity of aged oocytes was increased by the addition of EGCG. These suggest that EGCG can improve the quality of post-ovulatory aged oocytes.
Chia (Salvia hispanica L.) and quinoa (Chenopodium quinoa Willd.) are pseudocereals that stand out due to their high nutrient density and functional component content. Their richness in protein, dietary fiber, unsaturated fatty acids, vitamins, minerals, and bioactive phytochemicals makes them important dietary components with potential roles in metabolic processes and inflammation-related mechanisms. This study aimed to evaluate the effects of chia and quinoa seeds, included at 5%, 10%, and 20% in isocaloric and isonitrogenous diets, on serum adipokines (omentin, visfatin, resistin, chemerin), insulin, anthropometric measurements, and body mass index (BMI) in healthy, normal weight male Wistar rats, relative to a control diet. Seventy male Wistar albino rats were divided into seven groups: control diet, 5%, 10%, and 20% chia seed diets (LD-CD, MD-CD, HD-CD) and 5%, 10%, and 20% quinoa seed diets (LD-QD, MD-QD, HD-QD). After 6 weeks, serum adipokine and insulin levels were measured. Weekly body weight, body length, abdominal circumference, and chest circumference were recorded, and BMI values were calculated. Chia and quinoa supplementation altered serum adipokine and insulin levels, with the observed responses varying according to the supplementation level. High-dose (20%) supplementation decreased serum chemerin, resistin, and insulin levels while increasing omentin levels compared with controls. Moderate (10%) and high (20%) supplementation altered serum visfatin levels. Chia and quinoa supplemented diets reduced BMI compared with controls. The HD-CD group also showed lower body weight gain and abdominal circumference. Including chia and quinoa seeds as dietary alternatives may positively affect serum adipokine levels and anthropometric parameters.
Cisplatin, a commonly used chemotherapy drug, inevitably causes damage to the liver and kidneys while exerting its anti-tumor effects. Carboxymethylated Poria cocos polysaccharides (CMP) have anti-inflammatory, antioxidant, and organ-protective properties, and also exhibit significant anti-tumor activity. Therefore, this study aims to investigate the effects of CMP on cisplatin-induced hepatorenal damage in mice. The results showed that CMP treatment significantly mitigated abnormalities in hepatorenal function indicators and tissue damage induced by cisplatin. Furthermore, CMP significantly suppressed the increase in levels of inflammatory factors in peripheral and hepatorenal tissues, as well as the infiltration of immune cells induced by cisplatin. Additionally, CMP significantly enhanced the antioxidant capacity of hepatorenal tissues and inhibited oxidative damage caused by cisplatin. In vitro, CMP have also been shown to protect renal tubular epithelial cells and hepatocytes from cisplatin-induced damage. Mechanistically, our studies indicated that CMP activate Nrf2 through a phosphorylation pathway that is independent of Keap1 and reliant on PLC-PKC signaling. In conclusion, our findings suggest that CMP has the potential to serve as an adjunctive agent in cisplatin chemotherapy to protect the kidneys and liver from damage.
Ziziphora clinopodioides Lam. extract, as a natural bioactive compound, possesses a wide range of health benefits; however, its inherent instability limits practical applications. To overcome this limitation, nanoparticle-based delivery systems, particularly chitosan-gum arabic nanoparticles, were employed. The aim of this study was to evaluate the effects of varying concentrations of chitosan (3-9 mg/mL), Z. clinopodioides L. extract (3-9 mg/mL), and gum arabic (0.5-1.5 mg/mL), on the physicochemical characteristics of chitosan-gum arabic nanoparticles containing the extract, using a two-step emulsion-ionic gelation approach. Key parameters, including particle size, zeta potential, and encapsulation efficiency, were assessed by examining both individual and interactive effects. The process was optimized using response surface methodology (RSM) with a Box-Behnken design. The FTIR and XRD analyses were conducted to characterize the nanoparticles. Nanoparticle sizes ranged from 13.90 to 1722.75 nm, while the zeta potentials ranged from 1.63 to 46.67 mV, and the encapsulation efficiencies ranged from 20.51% to 63.66%. Notably, increasing the extract concentration reduced the encapsulation efficiency, while increasing particle size and zeta potential. Higher chitosan concentrations increased nanoparticle zeta potential. In contrast, increasing gum arabic concentration enhanced the encapsulation efficiency. The results demonstrated significant interaction effects among the formulation parameters across all measured responses. Optimal concentrations of chitosan, extract, and gum arabic were determined to be 9, 4.90, and 1.5 mg/mL, respectively, yielding a particle size of 50 nm, a zeta potential of 5.86 mV, and an encapsulation efficiency of 60.63%. The FTIR and XRD analyses confirmed proper encapsulation of the extract. These findings highlight the critical effects of studied parameters on practical characteristics of chitosan-gum arabic nanoparticles loaded with Z. clinopodioides L. extract.
Ulcerative colitis (UC) is a prevalent chronic inflammatory bowel disease globally, driven mainly by immune dysregulation and gut microbiota dysbiosis. This study focused on Lactiplantibacillus plantarum BF_15, an infant-derived strain with potential to alleviate UC. Four experimental groups were established, including the control group, DSS group, BF_15 group, and LGG group. Probiotics were administrated at a dosage of 1 × 109 CFU/mL and 0.1 mL/10 g body weight (BW) for 21 days. The therapeutic effects and mechanisms of BF_15 against UC were explored based on physiological indicators, gut microbiota, and tandem mass tag (TMT)-based quantitative proteomics. The results showed that BF_15 intervention significantly reduced the disease activity index (DAI) from 3.00 ± 0.50 to 1.16 ± 0.36 (p < 0.05) and mitigated UC-related symptoms including body weight loss and colon shortening, with colon length restored from 4.43 ± 0.32 cm to 5.41 ± 0.26 cm (p < 0.05). Meanwhile, BF_15 downregulated the levels of pro-inflammatory factors such as interleukin IL-6 and IL-17A, and upregulated the levels of IL-10 (from 565.92 ± 79.40 pg/mL to 656.84 ± 66.01 pg/mL) and secretory immunoglobulin A (SIgA, from 9.75 ± 1.29 μg/mL to 12.79 ± 1.77 μg/mL) (p < 0.05), with a therapeutic effect comparable to that of LGG. Further, gut microbiota analysis demonstrated that BF_15 restored the relative abundance of Bacteroidetes (from 28.02% to 30.85%) and regulated gut microbiota homeostasis. Finally, TMT-based proteomic analysis indicated that BF_15 regulated 528 differentially expressed proteins (DEPs) and exhibited a superior regulatory effect on the antigen presentation and processing (APC) pathway compared with LGG. The above findings provide potential novel therapeutic targets and probiotic strain resources for the treatment of UC.
This study examined how the inoculation sequence of Saccharomyces cerevisiae (Sc) and Wickerhamomyces anomalus (Wa), along with Dendrobium nobile Lindl. (D. nobile) addition, affects rice wine quality. Sequential inoculation with D. nobile yielded the highest levels of polysaccharides, alkaloids, flavonoids, and phenols. While simultaneous inoculation raised alcohol proportion, sequential inoculation increased ester content. D. nobile reduced total flavor concentration but enhanced its diversity. Metabolic analysis indicated that both factors alter metabolites mainly via amino acid and sugar pathways. Thus, sequential Sc-Wa inoculation with D. nobile is an effective strategy for modulating wine composition, providing a reference for improving fermented rice wine quality.
The appearance of the "oxidation band" in the fluorescence spectrum of anthocyanins is an indicator of oxidative deterioration of anthocyanins and can be a useful preliminary index of food freshness. Chokeberry (Aronia melanocarpa) fruit, rich in anthocyanins, can be subject to drying or lyophilization for domestic or industrial applications. The study was aimed at determining whether the measurement of fluorescence of dried chokeberry extracts can provide information on the extent of anthocyanin oxidation during prolonged drying at 60°C, and if fluorescence of fruit extracts can monitor anthocyanin oxidation during storage of the lyophilizates at various temperatures under air exposure. The increase in the fluorescence intensity of the extracts of chokeberry fruits or their lyophilizates at the "oxidation band" was a regular function of the time of fruit drying and the time of lyophilizate storage at 22°C and 37°C, confirming the usefulness of this parameter for estimation of oxidative changes in dried fruits and their lyophilizates. However, fluorescence of whole fruits, pulverized fruits and pulverized lyophilizate did not show a regular increase as a function of the time of fruit drying and lyophilizate storage, apparently due mainly to the effects of light scattering. The results point to the necessity of controlling the drying process of chokeberry fruits and storing their lyophilizates at ambient or higher temperatures with air access to avoid anthocyanin oxidation.
Orange-fleshed sweet potato (OFSP) is widely promoted in sub-Saharan Africa, including Senegal, for its contribution to vitamin A deficiency mitigation. However, its potential to improve mineral nutrition remains limited by low iron (Fe) and zinc (Zn) concentrations and their restricted bioavailability, which may contribute to persistent micronutrient deficiencies affecting 42% of children under 5 years and 37% of women of reproductive age in Senegal. In this context, organic waste products (OWPs) and microbial inocula (MIs) represent promising agroecological alternatives to conventional fertilizers for enhancing the nutritional quality of OFSP while valorizing locally available resources. This study aimed to evaluate the combined effects of various OWPs and MIs on tuber yield and Fe/Zn concentrations in OFSP within an agroecological production system. Field experiments were conducted across two contrasting seasons: a rainy season (June 2021-November 2021), followed by a dry season (December 2021-May 2022). The OFSP variety used was "Apomudem". Two types of MIs were tested: local beneficial microorganisms (BMs), a bacterial-fungal consortium derived from fermented forest litter, and mycorrhizal fungi (MF; Glomus mosseae). A factorial arrangement in a randomized complete block design with four replications was employed. Results revealed several combinations that significantly improved both yield and micronutrient content. The treatment combining poultry litter (PL) with BMs yielded 21 t ha-1 of tubers-a 2.6-fold increase compared to the unfertilized control-and doubled Zn concentration to 20 mg kg-1 DM. Additionally, the combination of PL, BMs, and MF increased Fe concentration to 44 mg kg-1 DM, a 1.7-fold increase. These effects varied with OWP and inoculum type, confirming that their interactions strongly influenced nutrient enrichment. Overall, combining OWPs with microbial inocula provides an effective agroecological approach for enriching crops with essential micronutrients and opens promising avenues for improving staple crop productivity and nutritional quality by effectively valorizing locally available resources, supporting agroecological transition, and food security.
Obesity causes chronic kidney damage. This study investigated the renoprotective effects of Gochujang, a Korean traditional fermented food, despite its high salt content. Gochujang extract (GE) significantly inhibited lipid accumulation and reduced inflammation-related proteins, including TNF-α, in renal tubular epithelial (HK-2) cells. Additionally, Gochujang markedly attenuated kidney injury-associated changes by lowering blood urea nitrogen (BUN) and kidney injury molecule-1 (KIM-1) levels, and by improving histopathology in high-fat diet (HD)-induced obese mice. Moreover, Gochujang altered HD-induced gut microbiota changes by increasing diversity and reducing the Firmicutes-to-Bacteroidetes ratio, whereas table salt had no effect. Importantly, alterations in the gut microbiota were strongly correlated with improvements in renal injury and inflammatory indicators, implying an association between the renoprotective effects of Gochujang and gut microbiota modulation. These findings highlight the potential of Gochujang to attenuate obesity-related kidney injury-related changes despite its high salt content.
This study aimed to optimize chlorophyll extraction from Ceratophyllum submersum L. and develop a stable spray-dried chlorophyll powder. Microwave-assisted extraction was optimized using Response Surface Methodology (Box-Behnken design) with MgCO3 concentration, microwave power, and extraction time as variables. The optimal conditions (0.763% MgCO3, 372.1 W, 5.28 min) yielded 0.187 mg/g dry mass, which closely matched the predicted value. Spray drying parameters were optimized at 25% maltodextrin, a feed rate of 6 mL/min, and an inlet temperature of 160°C. SEM revealed spherical particles with typical surface wrinkling; FTIR showed spectral features consistent with the chlorophyll-maltodextrin matrix, without allowing specific differentiation between chlorophyll and its degradation products, while XRD revealed a predominantly amorphous structure. The resulting powder exhibited a low moisture content (3.25%), high solubility (83.72%), and acceptable flowability, with a Carr index of 23.45% and a Hausner ratio of 1.30. The measured microbiological, heavy-metal, and residual-solvent levels met the evaluated safety criteria. These findings demonstrate the potential of C. submersum as an underutilized source of natural chlorophyll and provide laboratory-scale processing conditions for further compositional, storage-stability, application, and scale-up studies.
Clear cell renal cell carcinoma (ccRCC) is characterized by substantial metabolic and molecular heterogeneity, but the disease-relevant programs associated with acteoside, a dietary polyphenol, remain poorly understood. We integrated predicted acteoside targets with bulk, single-cell, and spatial transcriptomic data from ccRCC and combined molecular subtyping with cross-cohort machine-learning analysis. Acteoside-related signatures were preferentially enriched in malignant compartments and increased with tumor grade and stage. Consensus clustering identified two molecular subtypes with distinct biological and clinical features. C1 was associated with immune activation, metabolic activity, and more favorable survival, whereas C2 showed greater genomic instability, reduced renal epithelial differentiation, and poorer outcomes. We further benchmarked multiple machine-learning strategies and established a 10-gene prognostic model that retained predictive performance across independent cohorts, with IMPDH1 emerging as the strongest risk-associated feature. Functional experiments confirmed the biological relevance of IMPDH1: its knockdown suppressed ccRCC cell proliferation, DNA synthesis, colony formation, and migration, whereas overexpression produced the opposite effects. Together, these findings indicate that acteoside-related molecular signatures capture clinically relevant heterogeneity in ccRCC and provide a framework for linking dietary-polyphenol-related molecular space with tumor biology. The identification and functional validation of IMPDH1 further highlight its potential importance in ccRCC progression.
This study investigated a patented two-stage lyophilization technique to encapsulate phenolic extract from pomegranate peel using orange peel and commercial pectin as carrier materials. The aim was to improve the stability of bioactive compounds during storage and thermal processing in baked products. Pomegranate peel extract was encapsulated using two approaches: a two-stage lyophilization method with orange peel at a 7:1 extract-to-wall ratio and a conventional single-stage freeze-drying method with pectin. Encapsulated and non-encapsulated extracts were characterized by HPLC, evaluated for color stability during accelerated storage (60°C for 45 days), and incorporated into crackers (1000 ppm) to examine thermal stability, lipid oxidation, fiber enrichment, and sensory attributes. HPLC analysis identified 17-18 phenolic compounds across the different extracts, with punicalagin as the predominant phenolic compound. Compared with commercial pectin, the orange peel-based encapsulation system showed superior retention of phenolic compounds during storage. Following baking, phenolic loss was only 6.7% in the orange peel system, compared with 40.8% and 69.4% in the non-encapsulated and pectin systems, respectively. Encapsulation also improved color stability, reduced lipid oxidation, increased dietary fiber, and maintained sensory acceptability of fortified crackers. Overall, the patented orange peel-based encapsulation system demonstrated superior protection of pomegranate peel phenolic compounds compared with the conventional pectin-based system, highlighting the potential of citrus by-products as sustainable natural carrier materials for developing functional bakery products.
Osteoporosis remains a critical public health challenge worldwide, particularly for older adults. Dietary sodium intake is considered a potentially modifiable factor related to bone mineral density (BMD) and osteoporosis risk, but existing findings remain inconsistent, and large-scale evidence is limited. This study combines cross-sectional analysis from the National Health and Nutrition Examination Survey (NHANES) with Mendelian randomization (MR) to investigate the association and potential causal relationship between sodium intake and osteoporosis. Drawing on NHANES data from the 2005-2010, 2013-2014, and 2017-2018 cycles (n = 3350), we applied weighted multivariable logistic regression to evaluate the association between sodium intake and osteoporosis risk. Leveraging sodium metabolism-related single-nucleotide polymorphisms (SNPs) as instrumental variables, we performed two-sample Mendelian randomization (MR) analysis. MR estimates were derived using inverse-variance weighting (IVW), MR-Egger regression, and weighted median methods, with robustness assessed using pleiotropy tests and leave-one-out sensitivity analysis. Cross-sectional analysis showed that each 1 mg/day increase in two-day average dietary sodium intake was associated with approximately 0.011% lower odds of osteoporosis (OR = 0.99989, 95% CI: 0.99979-1.0000). Compared with the lowest quartile (Q1), participants in Q3 (OR = 0.48, 95% CI: 0.30-0.78, p = 0.004) and Q4 (OR = 0.51, 95% CI: 0.28-0.93, p = 0.028) had lower odds of osteoporosis (p-trend < 0.05). MR analysis using the IVW method provided genetic evidence supporting a potential inverse association (OR = 0.51, 95% CI: 0.28-0.95), although the MR-Egger estimate was nonsignificant and had a wide confidence interval (p = 0.312). These findings support a potential inverse association between sodium intake and osteoporosis risk in this study population. Further prospective and experimental studies are needed to validate these findings, clarify the underlying mechanisms, and determine whether they have any implications for dietary recommendations.
The objective of this study was to evaluate the effects of adding Jerusalem artichoke to the diet of fattening boars and the effects of polymorphisms in the CYP2E1 gene on skatole levels in back fat and feces and on CYP2E1 mRNA expression in pig liver. A total of 25 crossbreed boars (LWS × Pn) × (LD × LWD) were used in this study. All boars were genotyped for c.1423G > A. The pigs were divided into two different dietary treatment groups with 0% (C-control; n = 12) and 8.2% (E-experimental; n = 13) dried Jerusalem artichoke tubers, which were fed 14 days before slaughter. The results suggest that feeding Jerusalem artichoke decreased (p = 0.0236) the level of skatole in adipose tissue. In feces, a decrease in the skatole level was observed, but there was no statistically significant difference between the control and experimental groups. The influence of polymorphism on skatole levels in adipose tissue and feces were lowest for the AA genotype and highest for the GG genotype. Differences between genotypes for skatole levels in feces were statistically significant (p = 0.0242). Differences between genotypes in adipose tissue on skatole levels showed the same trend in favor of the A allele. However, these differences were not statistically significant. This study also confirmed the significant effect of Jerusalem artichoke on the liver CYP2E1 mRNA expression. The CYP2E1 gene expression level was significantly higher in the experimental group (p = 0.0086). No significant differences in mRNA expression between different genotypes were detected.
This study developed and optimized watermelon seed-based cream cheese (WCC) as a plant-based alternative to dairy cream cheese and compared its microstructure, volatile compounds, taste profiles, amino acid composition, and sensory characteristics with commercial vegan cream cheese (VC) and dairy cream cheese (DC). Microstructural analysis by scanning electron microscopy showed progressive network development from the prototype (P1) to the optimized formulation (P3), with increased aggregation and porosity associated with the incorporation of hydrocolloids, salt, and maltodextrin. GC-E-Nose analysis identified 30 volatile compounds across seven chemical groups and revealed significant differences among samples. P1 showed high levels of ethanol and aldehydes related to natural coagulation and lipid oxidation, whereas P2 and P3 showed reduced volatile intensities due to matrix stabilization and acidification. Dairy cream cheese exhibited characteristic fermentation-related volatiles, while VC showed the lowest volatile complexity. Electronic tongue analysis demonstrated significant differences in basic taste attributes among samples, with P3 showing higher saltiness and moderate sourness, shifting its taste profile toward VC. Amino acid analysis showed the highest contents in DC, whereas WCC samples were characterized by arginine-rich profiles and lower essential amino acid levels. Sensory descriptive analysis and PCA indicated that formulation optimization reduced seed-derived nutty and grain notes and increased milk-like attributes in P3. These findings demonstrate that formulation optimization improves the structural integrity and sensory similarity of WCC to commercial cream cheese.
Ulleungdo Island, Korea, features a distinctive oceanic climate that supports a diverse array of edible wild plants traditionally processed through blanching and sun-drying. Despite their cultural and commercial significance, the effects of traditional drying methods on phytochemical composition remain inadequately understood. This study investigated processing-induced metabolic changes in three representative Asteraceae species-Aster pseudoglehnii, Cirsium nipponicum, and Solidago virgaurea-using integrated metabolomics and machine learning approaches. Untargeted UHPLC-HRMS/MS profiling identified 110 secondary metabolites across freeze-dried and sun-dried samples. Hierarchical clustering revealed that species identity was the primary determinant of chemotype, while processing induced consistent quantitative shifts within each species. Traditional processing (blanching followed by sun-drying) significantly reduced photo- and enzymatically labile flavonoid glycosides, whereas caffeoylquinic acid derivatives were relatively preserved. Random Forest analysis combined with SHAP interpretation identified caffeoylquinic acids, including chlorogenic acid and dicaffeoylquinic acid isomers, as processing-stable chemotaxonomic markers, while glycosylated flavonoids served as sensitive indicators of processing intensity. Targeted quantification confirmed these trends and revealed species-specific phenolic allocation patterns. Overall, this study demonstrates that integrating metabolomics with explainable machine learning enables robust identification of processing-dependent marker compounds and provides a biochemical foundation for traditional drying practices and marker-based quality control strategies for Ulleungdo wild vegetables.
Cardiorenal anemia syndrome (CRAS) links heart failure, chronic kidney disease, and anemia, but the role of diet-related inflammation remains unclear. In the National Health and Nutrition Examination Survey (NHANES), we examined associations of the Dietary Inflammatory Index (DII) with CRAS, dose-response patterns, inflammatory mediation, and machine-learning prediction, alongside analysis of a myocardial infarction-CKD-anemia phenotype in the Korea National Health and Nutrition Examination Survey (KNHANES). This study included 25,388 NHANES and 32,935 KNHANES adults. Survey-weighted regression and restricted cubic splines assessed associations with CRAS in NHANES and the related phenotype in KNHANES. Exploratory mediation evaluated six inflammatory indices. In NHANES participants with heart failure, least absolute shrinkage and selection operator regression selected predictors for 12 internally validated models, with cross-phenotype application in KNHANES and interpretation using Shapley additive explanations (SHAP). Higher DII was associated with greater odds of CRAS in NHANES (odds ratio [OR] = 1.22, 95% CI: 1.07-1.40) and of the related phenotype in KNHANES (OR = 1.19, 95% CI: 1.10-1.29). The association was largely linear in NHANES but nonlinear in KNHANES. In NHANES, all six indices showed positive indirect associations, with estimated proportions ranging from 5.22% to 16.26%. CatBoost showed the highest discrimination in NHANES internal validation (ROC-AUC = 0.741; PR-AUC = 0.556) and in the KNHANES cross-phenotype application (ROC-AUC = 0.753; PR-AUC = 0.698). Higher DII was associated with greater odds of CRAS in NHANES, with directionally consistent findings for the related phenotype in KNHANES. The model may support model-based CRAS identification and showed preliminary applicability to the related phenotype.
Anthocyanin accumulation in Rosaceae fruits is governed by a multi-layered regulatory network integrating transcriptional control, epigenetic modulation, environmental signaling, and metabolic flux allocation. This review presents a critical synthesis of recent advances in this field, moving beyond descriptive gene lists to examine the mechanistic underpinnings of pigment regulation. We first outline the core biosynthetic pathway and post-synthetic modifications that ensure pigment stability. The central focus is on the MYB-bHLH-WD40 transcriptional complex and its integration with epigenetic mechanisms (DNA methylation, histone modifications, non-coding RNAs) and post-transcriptional regulation. We further examine how environmental cues-light, temperature, and hormones-are transduced through these molecular networks, and analyze metabolic flux control at pathway branch points. A cross-species comparative perspective across major Rosaceae crops (Malus, Pyrus, Fragaria, Prunus) reveals conserved regulatory nodes and genus-specific innovations. Finally, we discuss how this integrated knowledge, combined with multi-omics and gene editing tools, is driving targeted strategies in molecular breeding and postharvest quality management. This framework provides a foundation for rational manipulation of fruit coloration and nutritional quality.
The present study reports the green synthesis of iron oxide nanoparticles (IONPs) using the aqueous extract of Crocus sativus (saffron) as a natural reducing and stabilizing agent, followed by comprehensive in vitro and in vivo biological investigations. IONPs were confirmed by UV-visible spectroscopy (peak at 430 nm), FTIR analysis (peaks at 3400, 2900, 1650, 1380, and 1050 cm-1 indicating polyphenols, flavonoids, and proteins), and SEM imaging, which revealed a highly aggregated, irregular morphology. ImageJ analysis features gave a mean equivalent diameter of 111.98 ± 83.70 nm. EDS analysis confirmed iron (44.74 wt%) and oxygen (24.82 wt%) as the major elements with organic capping (28 wt% carbon). The phytochemical constituents of C. sativus are rich in phenolics and flavonoids, which effectively facilitated the reduction of Fe3+ ions to stable nanoscale iron oxide particles. The in vitro enzyme inhibition assays revealed significant inhibitory effects on α-glucosidase and xanthine oxidase, with IC50 values of 33.08 ± 1.29 and 2.96 ± 0.49 μg/mL, respectively, and moderate inhibition of carbonic anhydrase-II (49%). Antibacterial evaluation demonstrated an important zone of inhibition ZOI (21.45 mm) against Klebsiella pneumoniae, indicating the antimicrobial potential of the IONPs compared to the crude extract. In vivo pharmacological screening further supported their biological efficacy. IONPs showed potent analgesic activity (78% inhibition at 10 mg/kg), pronounced anti-inflammatory effects in carrageenan- and histamine-induced paw edema models (up to 95% inhibition), and marked sedative properties at low doses. These results suggest that Crocus sativus-mediated IONPs may possess strong therapeutic potential due to their enhanced biochemical, antimicrobial, anti-inflammatory, and analgesic activities. The study highlights saffron-assisted green synthesis as an efficient, eco-friendly, and biocompatible approach for developing multifunctional nanomaterials with possible applications in pharmaceutical, biomedical, and health-related fields.
Grilled food toxicants 4-hydroxynonenal (4-HNE), benzo[a]pyrene (BaP), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) pose health risks, including non-alcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC). However, their synergistic molecular mechanisms in driving NASH to NASH-HCC progression remain unclear. We employed network toxicology (CTD, GeneCards), bioinformatics analysis of the GSE164760 dataset, machine learning (LASSO, SVM-RFE, Random Forest), SHAP interpretability analysis, immune infiltration profiling (CIBERSORT), miRNA-TF-mRNA network construction, and molecular docking to identify core genes and elucidate mechanisms. Toxicity prediction confirmed significant hepatotoxicity for all three compounds. Intersection analysis identified 8 key genes (InterGenes) enriched in oxidant detoxification, fatty acid metabolism, and chemical carcinogenesis pathways. Machine learning refined this to 6 CoreGenes (GSTA1, EPHX1, CYP2E1, GPX3, ALB, TF). SHAP analysis revealed that high expression of ALB, TF, GSTA1 and CYP2E1, together with low expression of GPX3 and EPHX1, correlated with increased progression risk. A CoreGenes-based diagnostic nomogram achieved exceptional performance (AUC = 0.967). Immune infiltration analysis revealed significant shifts in regulatory T cells and M2 macrophages during progression. Molecular docking confirmed strong binding affinities between the toxicants and CoreGenes proteins (e.g., BaP-GSTA1: -10.7 kcal/mol). Functional enrichment implicated dysregulated fatty acid metabolism, PPAR, and TGF-beta signaling. This study identifies GSTA1, EPHX1, CYP2E1, GPX3, ALB, and TF as pivotal biomarkers and mediators through which 4-HNE, BaP, and PhIP synergistically promote NASH-HCC progression via oxidative stress, metabolic dysfunction and remodeling of the immune microenvironment. The CoreGenes signature and multi-gene model provide a powerful tool for risk assessment and early intervention.
