Phenylketonuria represents the most prevalent inborn error of amino acid metabolism. In early diagnosed patients adequate and continued dietary treatment results in a good neurologic outcome. However, due to the natural protein and phenylalanine-restricted diet, oxidative stress represents a concern in phenylketonuric patients. Clear evidences suggest that the pathophysiology of PKU is also dependent by mitochondrial impairment and oxidative stress. In this context due to the tight connection between oxidative and inflammatory stress and noncommunicable diseases (NCDs) development, it is reasonable to hypothesize that PKU patients may present a higher risk to develop NCDs during their life. Currently available protein substitutes on the market include free amino acids (L-AAs), prolonged-release protein substitute and formula containing glycomacropeptide (GMP). Our results suggest that free L-AAs significanlty worsens the intestinal hydrogen peroxide (H₂O₂₎ and lipopolysaccharides (LPS)-induced oxidative and inflammatory status in Caco-2 cells, which are significantly restored towards physiological condition by GMP alone and when present in a 1:1 mixture with free L-AAs, providing new preclinical piece of information which can shed a shadow on the mechanism of action of these products on PKU patients and their future management.

In recent years, there has been growing interest in exploring alternative and innovative delivery systems to improve the efficacy of iron supplements, satisfying iron needs and lowering side effects. To address this issue, this study aimed at demonstrating the advantages of Ferro Supremo formulation (composed of encapsulated iron, vitamins, and micronutrients), in terms of capacity to improve iron intestinal absorption, in comparison with standard FeSO₄. Hence, differentiated Caco-2 cells have been used for assessing the in vitro bioavailability and safety of FS and FeSO₄. MTT experiments demonstrated that both FS and FeSO₄ are not able to impair the viability of Caco-2 cells. Furthermore, the quantitative and qualitative analysis, conducted by atomic absorption spectrometry and fluorescence determinations, revealed that FS can enter, accumulate in the cytoplasm, and be transported by intestinal cells four times more efficiently than FeSO₄. Our findings indicate that this formulation can be considered a valuable and efficiently good choice as food supplements for improving iron deficiency.

This study demonstrates the effectiveness of ultrasonication, as a pre-treatment technology, coupled to enzymatic hydrolysis of herring side streams, yielding multifunctional peptide mixtures with antioxidant, hypotensive (ACE inhibitory activity), and hypoglycemic (DPP-IV inhibitory and GLP-1 enhancer activity) properties. The ultrasound pre-treatment modulates the biological activity of the hydrolysates, enhancing certain bioactive properties (antioxidant, ACE inhibitory, and GLP-1 enhancer activities, respectively) while reducing others (DPP-IV inhibitory activity). The study also highlights the importance of simulating gastrointestinal digestion and using Caco-2 cells to assess the bioaccessibility, intestinal bioavailability, and metabolic resistance of herring peptides. These findings support the use of ultrasonication and enzymatic hydrolysis in obtaining multifunctional bioactive peptide mixture for the prevention of metabolic syndrome. Results clearly suggest that this approach represent sustainable solutions in food science and technology, since it allowed us to obtain a bioactive mixture of peptides starting from fish by-products pre-treated with green methodologies.

Purpose: Docosahexaenoic acid (DHA) is a long-chain omega‐3 polyunsaturated fatty acid. We investigated the dual health ability of DHA to modulate gut microbiota in children with obesity and to exert anti-inflammatory activity on human intestinal Caco-2 cells. Methods: In a pilot study involving 18 obese children (8–14 years), participants received a daily DHA supplement (500 mg/day) and dietary intervention from baseline (T0) to 4 months (T1), followed by dietary intervention alone from 4 months (T1) to 8 months (T2). Fecal samples, anthropometry, biochemicals and dietary assessment were collected at each timepoint. At preclinical level, we evaluated DHA’s antioxidant and anti-inflammatory effects on Caco-2 cells stimulated with Hydrogen peroxide (H₂O₂) and Lipopolysaccharides (LPS), by measuring also Inducible nitric oxide synthase (iNOS) levels and cytokines, respectively. Results: Ten children were included in final analysis. No major changes were observed for anthropometric and biochemical parameters, and participants showed a low dietary compliance at T1 and T2. DHA supplementation restored the Firmicutes/Bacteroidetes ratio that was conserved also after the DHA discontinuation at T2. DHA supplementation drove a depletion in Ruminococcaceae and Dialisteraceae, and enrichment in Bacteroidaceae, Oscillospiraceae, and Akkermansiaceae. At genus level, Allisonella was the most decreased by DHA supplementation. In Caco-2 cells, DHA decreased H₂O₂-induced reactive oxygen species (ROS) and nitric oxide (NO) production via iNOS pathway modulation. Additionally, DHA modulated proinflammatory (IL-1β, IL-6, IFN-γ, TNF-α) and anti-inflammatory (IL-10) cytokine production in LPS-stimulated Caco-2 cells. Conclusion: An improvement in gut dysbiosis of children with obesity seems to be triggered by DHA and to continue after discontinuation. The ability to modulate gut microbiota, matches also with an anti-inflammatory effect of DHA on Caco-2 cells. Graphical