Mycotoxins, secondary metabolites produced by molds, pose significant health risk through contamination of globally consumed cereals. Ochratoxin A (OTA), a prevalent mycotoxin in cereals, is associated with various health hazards, including immunotoxicity. This study explores the bioaccessibility of OTA in bread and its impact on the gastrointestinal barrier. A focus is placed on grape pomace (GP), a by-product of the wine industry, as a potential mitigator of OTA toxicity. Results demonstrate that GP reduces OTA bioaccessibility in the human gastrointestinal system from 94% to 81% at intestinal level, showing promise in limiting the absorption of the harmful toxin. Additionally, GP exhibits cytoprotective effects, enhancing cell viability and mitigating OTA-induced toxicity in both Caco-2 and Jurkat T cells. In view of the above, to understand the mechanisms by which OTA exhibits its toxic effects, flow cytometry was chosen as the main technique for the analysis of cell cycle, reactive oxygen species levels and mitochondrial parameters. Cytofluorimetric evaluation indicates GP's potential in limiting OTA-induced damage at cellular level. The study suggests that GP could serve as functional ingredient to reduce mycotoxin bioaccessibility and toxicity in cereal-based foods, offering a novel and promising approach to enhance food safety and protect public health. The finding highlights the potential of utilizing grape pomace in food formulations to mitigate mycotoxin contamination, providing a valuable contribution to the ongoing efforts to ensure the safety of globally consumed cereal products.

Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus ochraceous and various Penicillium species, which are known for contaminating agricultural products and posing significant health risks, which include immunotoxicity. This study aims to evaluate the potential of nanostructured lipid carriers (NLCs) loaded with a carotenoid-enriched extract from pumpkin peel (Cucurbita maxima L.) in mitigating the toxic effects of OTA. To address the poor bioavailability and instability of carotenoids, nanoencapsulation techniques were employed to enhance their delivery and efficacy. NLCs were formulated using hydrogenated sunflower oil, pumpkin oil, and soy lecithin using hot high-pressure homogenization. The in vitro study involved co-digesting OTA-contaminated bread with an NLC formulation and assessing the impact of the encapsulated carotenoid on OTA bioaccessibility, bioavailability, and cellular toxicity using Caco-2 and Jurkat T cells. Even though no significant influence was observed on the bioaccessibility and bioavailability of OTA, carotenoid-loaded NLCs exhibited cytoprotective effects by improving cell viability and mitigating OTA-induced toxicity in both Caco-2 and Jurkat T cells. Particularly, the flow cytometry analysis highlighted the ability of carotenoids to mitigate OTA-induced cellular damage by decreasing ROS production and limiting mitochondrial mass changes. The study suggests that the encapsulation of carotenoids in NLCs represents a promising strategy to enhance their protective effects against OTA toxicity, potentially offering a novel approach to food safety and public health protection. The study underscores the potential of nanotechnology in improving the bioavailability and efficacy of natural antioxidants to mitigate mycotoxin-induced damage.

Mycotoxin contamination of cereals and cereal-based products is a serious problem for food safety. Antioxidant-rich ingredients such as bilberries (Vaccinium myrtillus L., VM) may mitigate their harmful effects. Firstly, total phenolic content, antioxidant activity, and analytical phytochemical composition (hydroxycinnamic and hydroxybenzoic acids, flavanols, flavonols, and anthocyanins) were assessed in lyophilized wild bilberries from Romania. Secondly, this study evaluated bilberries’ effects on reducing ochratoxin A (OTA) bioaccessibility and cytotoxicity. An in vitro digestion model was developed and applied to four different types of bread: Control, VM (2%), OTA (15.89 ± 0.13 mg/kg), and OTA (16.79 ± 0.55 mg/kg)-VM (2%). The results indicated that VM decreased OTA bioaccessibility by 15% at the intestinal level. OTA-VM digests showed improved Caco-2 cell viability in comparison to OTA digests across different exposure times. Regarding the alterations in Jurkat cell line cell cycle phases and apoptosis/necrosis, significant increases in cell death were observed using OTA digests (11%), while VM addition demonstrated a protective effect (1%). Reactive oxygen species (ROS) analysis confirmed these findings, with OTA-VM digests showing significantly lower ROS levels compared to OTA digests, resulting in a 3.7-fold decrease. Thus, bilberries exhibit high potential as a functional ingredient, demonstrating protection in OTA mitigation effects.

Background: Vaccinium berries are rich in polyphenolic compounds with recognized health properties; however, studies integrating phytochemical characterization with functional cellular assays remain limited. This study comparatively evaluated wild (Vaccinium myrtillus L., VM) and cultivated blueberry (Vaccinium corymbosum L., VC) using phytochemical profiling alongside in vitro antioxidant and anti-inflammatory activity testing. Six samples from Romanian regions were extracted using ultrasound-assisted methods, then analyzed for total phenolics, flavonoids, tannins, and antioxidant activity (DPPH, ABTS, FRAP, CUPRAC). Anthocyanins were further characterized by LC–MS/MS. The most bioactive extracts were tested for cytocompatibility, intracellular ROS modulation in Caco-2 cells, and anti-inflammatory activity in LPS-stimulated THP-1 cells. VM extracts showed higher total phenolic, anthocyanin contents, and antioxidant capacity than VC, with LC–MS/MS confirming a richer anthocyanin profile and differences in other phenolic compounds. Both extracts significantly reduced intracellular ROS in H₂O₂-stimulated Caco-2 cells and decreased IL-8 secretion in LPS-stimulated THP-1 cells, with a more pronounced dose-dependent anti-inflammatory effect for VM. These findings demonstrate that phytochemical differences between wild and cultivated Vaccinium species translate into distinct biological activities at the cellular level. The integrated approach provides a mechanistic link between composition and function, supporting the superior antioxidant and anti-inflammatory potential of VM and its relevance for functional food and nutraceutical applications.

Background/Objectives: Overweight and obesity are increasing globally. However, structured contemporary data on lifestyle behaviors and adiposity in Romanian young adults remain limited. Therefore, this study aimed to describe dietary and lifestyle habits, BMI, and overweight/obesity prevalence in Romanian adults aged 18–30 years and to examine associations between these variables. Methods: This cross-sectional online questionnaire study included 1202 young Romanian adults. BMI was calculated from self-reported height and weight and analyzed continuously, as well as for overweight/obesity (BMI ≥ 25 kg/m²). Pre-specified exposures were compulsive eating, soft-drink intake, breakfast frequency, physical activity, and sleep duration. Multivariable linear regression with heteroscedasticity-consistent standard errors was used for BMI, and modified Poisson regression with robust variance was used for overweight/obesity. Composite dietary score, sex-interaction, and sensitivity analyses were also performed. Results: Mean age was 23.2 ± 3.3 years, mean BMI was 23.8 ± 4.2 kg/m², and 32.4% of participants had overweight/obesity. Men had higher BMI and a higher prevalence of overweight/obesity than women. Compulsive eating and soft-drink intake showed dose-dependent associations with higher BMI and higher overweight/obesity prevalence. Short sleep duration (≤5 h/night) and daily breakfast consumption were associated with a higher and, respectively, lower prevalence of overweight/obesity. Physical activity showed no independent association after full adjustment, although this finding may be influenced by the use of a single self-reported item. Composite-score analyses supported the main findings. Conclusions: In Romanian young adults, compulsive eating and soft-drink intake were the most consistent behavioral correlates of adiposity, while breakfast regularity and short sleep showed threshold-type associations with overweight/obesity. These findings may inform the design of multicomponent prevention strategies, although longitudinal confirmation is needed.

Background/Objectives: Mycotoxin contamination in grain-derived foods is still a major food safety concern; thus, innovative mitigation approaches need to be continuously developed. This study investigated the influence of bilberry (Vaccinium myrtillus L.) incorporated into a food matrix on ochratoxin A (OTA)-induced cellular responses using a dietary-relevant in vitro intestinal model. Methods: Four bread types were prepared: control (C), OTA-contaminated (OTA), bilberry-enriched (VM), and OTA + VM (OTA-VM). Simulated intestinal digests of these breads were applied to differentiated Caco-2 cells for 24 h. Apoptotic and necrotic cell populations, as well as reactive oxygen species (ROS) levels, were quantified by flow cytometry, while RT-qPCR assessed the expression of 10 genes related to mitochondrial function, oxidative stress response, and intestinal barrier integrity. Results: Exposure to OTA resulted in increased cytotoxicity, reflected by a higher proportion of necrotic cells (5.11 ± 0.35%), and elevated ROS levels compared with control cells. Co-exposure to bilberry-enriched digests was associated with attenuation of apoptotic responses, a reduced proportion of necrotic cells (2.16 ± 0.61%) and a 16% decrease in ROS levels. Gene expression profiles in the VM group were comparable to control, whereas OTA exposure led to downregulation of several genes related to oxidative stress response and intestinal barrier integrity (e.g., CLDN2, OCLN, SLC7A11). In the OTA-VM group, a partial recovery of gene expression was observed. Conclusions: These findings suggest that bilberry incorporation into a food matrix may modulate OTA-induced cellular stress responses by attenuating oxidative imbalance and supporting the expression of genes associated with antioxidant defense and epithelial barrier integrity. Bilberries may therefore represent a promising functional ingredient for influencing intestinal cellular responses to dietary mycotoxin exposure.