The production of plant helical virus-like particles (VLPs) via plant-based expression has been problematic with previous studies suggesting that an RNA scaffold may be necessary for their efficient production. To examine this, we compared the accumulation of VLPs from two potexviruses, papaya mosaic virus and alternanthera mosaic virus (AltMV), when the coat proteins were expressed from a replicating potato virus X- based vector (pEff) and a non-replicating vector (pEAQ-HT). Significantly greater quantities of VLPs could be purified when pEff was used. The pEff system was also very efficient at producing VLPs of helical viruses from different virus families. Examination of the RNA content of AltMV and tobacco mosaic virus VLPs produced from pEff revealed the presence of vector-derived RNA sequences, suggesting that the replicating RNA acts as a scaffold for VLP assembly. Cryo-EM analysis of the AltMV VLPs showed they had a structure very similar to that of authentic potexvirus particles. Thus, we conclude that vectors generating replicating forms of RNA, such as pEff, are very efficient for producing helical VLPs.

: In the rapidly evolving field of nanotechnology, plant virus nanoparticles (pVNPs) are emerging as powerful tools in diverse applications ranging from biomedicine to materials science. The proteinaceous structure of plant viruses allows the capsid structure to be modified by genetic engineering and/or chemical conjugation with nanoscale precision. This means that pVNPs can be engineered to display peptides and proteins on their external surface, including immunodominant peptides derived from pathogens allowing pVNPs to be used for active immunization. In this context, pVNPs are safer than VNPs derived from mammalian viruses because there is no risk of infection or reversion to pathogenicity. Furthermore, pVNPs can be produced rapidly and inexpensively in natural host plants or heterologous production platforms. : In this review, we discuss the use of pVNPs for the delivery of peptide antigens to the host immune in pre-clinical studies with the final aim of promoting systemic immunity against the corresponding pathogens. Furthermore, we described the versatility of plant viruses, with innate immunostimulatory properties, in providing a huge natural resource of carriers that can be used to develop the next generation of sustainable vaccines.

Allergic disease diagnosis is currently experiencing a breakthrough due to the use of allergenic molecules in serum-based assays rather than allergen extracts in skin tests. The former methodology is considered a very innovative technology compared with the latter, since it is characterized by flexibility and adaptability to the patient’s clinical history and to microtechnology, allowing multiplex analysis. Molecular-based analysis requires pure allergens to detect IgE sensitization, and a major goal, to maintain the diagnosis cost-effective, is to limit their production costs. In addition, for the production of recombinant eukaryotic proteins similar to natural ones, plant-based protein production is preferred to bacterial-based systems due to its ability to perform most of the post-translational modifications of eukaryotic molecules. In this framework, Plant Molecular Farming (PMF) may be useful, being a production platform able to produce complex recombinant proteins in short time-frames at low cost. As a proof of concept, PMF has been exploited for the production of Bet v 1a, a major allergen associated with birch (Betula verrucosa) pollen allergy. Bet v 1a has been produced using two different transient expression systems in Nicotiana benthamiana plants, purified and used in a new generation multiplex allergy diagnosis system, the patient-Friendly Allergen nano-BEad Array (FABER). Plant-made Bet v 1a is immunoreactive, binding IgE and inhibiting IgE-binding to the Escherichia coli expressed allergen currently available in the FABER test, thus suggesting an overall similar though non-overlapping immune activity compared with the E. coli expressed form.

Introduction: Current in vitro intestinal models lack the mechanical forces present in the physiological environment, limiting their reliability for nanotoxicology studies. Here, we developed an enhanced Caco-2/HT29-MTX-E12 co-culture model incorporating orbital mechanical stimulation to better replicate intestinal conditions and investigate nanoparticle interactions.Methods: We established co-cultures under static and dynamic conditions, evaluating their development through multiple approaches including barrier integrity measurements, gene expression analysis, and confocal microscopy. We introduced novel quantitative analysis of dome formation as a differentiation marker and demonstrated the model application by investigating cellular responses to titanium dioxide (TiO₂) nanoparticles in a digested food matrix.Results: Dynamic conditions accelerated epithelial differentiation, achieving functional barrier properties by day 14 rather than day 21, with enhanced mucin production and more organized three-dimensional structure. Mechanical stimulation selectively promoted goblet cell differentiation without affecting general epithelial markers. The optimized model successfully detected concentration-dependent oxidative stress responses to TiO₂ exposure, revealing cellular dysfunction preceding membrane damage.Discussion: This improved co-culture system provides a better physiological platform for nanotoxicology studies. By incorporating mechanical forces, each cell type exhibits more representative behavior, creating a more realistic experimental setup. The model bridges the gap between simple monocultures and complex 3D systems, offering a practical approach for investigating nanoparticle-epithelium interactions in a food-relevant context.

Despite growing interest in ancient wheat varieties, the functional and nutritional properties of einkorn (Triticum monococcum) in cereal-based foods remain not fully elucidated. This study examined the chemical composition and functional properties of wholegrain einkorn pasta through cooking and simulated gastrointestinal digestion, comparing it with conventional Triticum durum wheat pasta. While sharing similar macronutrient profiles, einkorn pasta demonstrated higher retention of key compounds including phenolics, tocopherols, and phytosterols throughout cooking and in vitro digestion. Notable findings include enhanced prebiotic activity specifically targeting bifidobacteria populations and preserved antioxidant capacity despite thermal processing. These results demonstrated einkorn’s potential as a functional food ingredient, suggesting its capacity to deliver enhanced nutritional benefits through its unique matrix properties. Our findings provide mechanistic insights into ancient grain functionality in modern food applications, with implications for developing nutritionally enhanced pasta products.

Ferroptosis, an iron-dependent form of regulated cell death, has recently emerged as a crucial process in the pathogenesis of Pyricularia oryzae, the causal agent of the devastating rice blast disease, which causes billions of dollars in annual losses. This mini review explores the potential of antioxidants in suppressing ferroptosis in P. oryzae to promote sustainable rice production, with significant implications for global food security and nutrition. We critically analyze the current literature on the mechanisms of ferroptosis in P. oryzae, including iron metabolism and lipid peroxidation, the role of different antioxidants in inhibiting this cell death pathway, and the potential applications of antioxidant-based strategies for the management of rice blast disease. Recent discoveries, such as the efficacy of the natural flavonoid tangeretin in inhibiting fungal ferroptosis by interfering with the accumulation of iron and reactive oxygen species, highlight the promise of natural and nature-inspired compounds for disease management. The use of antioxidants to modulate ferroptosis in P. oryzae offers several advantages over traditional fungicide-based approaches, including improved safety, sustainability, and potential nutritional benefits through antioxidant-enriched rice varieties. However, challenges such as optimizing delivery methods, managing potential resistance, and ensuring efficacy under different environmental conditions need to be addressed. To achieve these goals, future research should focus on identifying the most effective antioxidant compounds, exploring synergistic combinations, and developing sustainable application methods.