The effects of Perilla frutescens pollination on the content of seed antioxidants were analysed by agronomical and pollination trials, comparing seeds produced from bagged plants in 2013 (A) to prevent access to pollinating insects, and seeds from open-pollinated plants in 2013 (B) and 2015 (C). The seeds of open-pollinated plants were significantly more numerous and heavier than those of self-pollinated plants. ¹H NMR seed analysis showed a higher presence of phenolic compounds in open-pollinated seeds, mainly rosmarinic acid and flavonoids, apigenin and luteolin. Flavonoids were present in the glucosylated form in seeds (A) and (C), and in the aglycone form in seeds from (B) plants. Saturated and unsaturated fatty acids (palmitic, linoleic and linolenic) were more abundant in seeds from self-pollinated flowers. Pollination performed almost exclusively by the honeybee notably increased the antioxidant content in perilla seeds and gave rise to a reduction in the fatty acid content.

The screening of plant-derived compounds with anti-cancer properties is a promising strategy to meet the growing need for new, safe and effective anti-cancer drugs. Justicidin B is a plants secondary metabolite that displays anti-cancer properties in several tumor cells. Therefore, it represents a good candidate. We used the 3R-compliant organism Caenorhabditis elegans to evaluate the safety of justicidin B produced by in vitro-grown adventitious roots of Linum lewisii. We showed that a dose of 100 µg/mL justicidin B does not affect worm vitality in either short-term or chronic administration; in contrast, the 200 µg/mL dose induces a lifespan reduction, but only in short-term daily treatment. We attributed this effect to its accumulation in lipofuscin granules in the pharynx as observed through confocal analysis. HPLC analysis confirmed the higher accumulation justicidin B with a 200 µg/mL dose but also revealed the presence of metabolic derivatives that could be responsible for the toxicity. We also demonstrated that the 100 µg/mL dose does not affect worm fertility or development. Our results highlight the safety of justicidin B, supporting its employment in cancer therapy, and encourage the use of a C. elegans model as an appropriate tool to assess compounds’ toxicity before moving to more complex organisms.

The use of natural bioactive compounds in edible coatings provides a sustainable approach to reducing food spoilage and meeting consumer demand for safer food preservation. In this study, bioactive extracts from Brassica juncea (green mustard, GM) and Raphanus sativus (radish tango, RT) sprouts were encapsulated into zein/chitosan (Z/CH) microparticles (MPs) using a complex coacervation–based encapsulation approach. The encapsulated microparticles (MPs), characterized by FTIR and UV-Vis spectroscopy, demonstrated a high loading efficiency of up to 90% and maintained their antioxidant activity for up to 168 h. TGA and SEM tests confirmed that the edible films produced by incorporating these microparticles (MPs) into polyvinyl alcohol (PVA) and chitosan (CH) matrices had a more uniform microstructure and enhanced heat stability. The Z/CH/RT6:PVA (1:2) and Z/CH/GM6:CH (1:1) formulations of the films showed significant antioxidant and antibacterial action, with up to 22.4% DPPH inhibition and a 1-log decrease in Salmonella enterica CFU, respectively. Overall, the results underscore the promise of sprout-derived microparticles as components for developing active, biodegradable packaging films with improved functional properties.