## 4 Discussion  
Bioactive peptides from fermented foods have recently attracted attention for their antioxidant properties [31; 32]. Peptides are fragments nascent in the primary protein sequences and could impart health benefits beyond basic nutritional advantages. It has been suggested that the amino acid composition can determine the radical scavenging, inhibition of lipid peroxidation, and metal ion chelation activity of peptides [33]. However, antioxidant and anti-inflammatory activities are interdependent since an oxidant stimulus can induce inflammation. Inflammatory cells liberate a number of reactive species at the site of inflammation, leading to oxidative stress; on the other hand, a number of reactive oxygen species can induce an intracellular signaling cascade that enhances the expression of proinflammatory genes [34]. Figure 5: Western blot analysis (on the left) and relative protein levels (on the right) from cells treated with LMW peptides from breads. RAW 264.7 cells were incubated with LMW peptides (0.05 mg/mL) from the breads for 1 h and then with LPS for 24 h. (**a**) The intensity of the signal from anti-phospho-NFkB mAbs was normalized on the anti-total NFkB mAbs signal. (**b**) anti-IkB and (**c**) anti-COX2 mAbs signals were normalized on the actin signal. NT: no treated cells; LPS: cells treated with lipopolysaccharide; ASA: cells treated with LPS and acetyl salicic acid. Aubusson, Rondo, and Skorpion indicate cells treated with both LPS and LMW peptides. Data reported in the figure represent the mean (\(\pm\)SD) of three different experiments performed in duplicate. Statistical analysis was performed for each sample vs. LPS. (* \(p\) < 0.05; *** \(p\) < 0.01; *** \(p\) < 0.001). Therefore, in order to define a food-derived molecule as a nutraceutical, it should at least have antioxidant and anti-inflammatory properties. In this respect, bioactive peptides are good candidates, since they often show the ability to regulate several inflammatory markers, such as inducible nitric oxide synthase (iNOS), cyclooxygenase2 (COX-2), cytokines, and chemokines, that act in concert with NF-kB activation and IkB degradation on the inflammation pathway [23; 35; 36]. Healthy breads are defined as baked products that can have beneficial effects on human wellness, and sourdough fermentation greatly increases the sensory properties, shelf-life, and nutritional value of the products, and it naturally induces the production of bioactive peptides equipped with antioxidant and anti-inflammatory properties [1]. In particular, some LAB strains have been shown to produce doughs with greater functional and nutritional value than others and bioactive peptides derived from the doughs have been characterized [8; 10; 37]. Most of the peptides produced during fermentation are derived from the main proteins of the flour [10]. Therefore, it is of peculiar importance to select flours as well as LAB and yeast strains to obtain breads with high contents of functional compounds, such as polyphenols, free amino acids and derivatives, and bioactive peptides [5; 37; 38; 39]. Previous studies have been performed on sourdoughs obtained with whole grain flour, including a hull-less barley with high \(\beta\)-glucan content, a blue-grain wheat flour (cv Skorpion) containing high anthocyanin concentrations, and a conventional red-grained wheat variety (cv Aubusson). They showed that barley sourdough possessed a strong antioxidant activity, generally higher than Skorpion samples, that, however, was greater than that recovered in Aubusson doughs [5]. Starting from these observations, these flours were used for the preparation of sourdoughs and breads. Three types of sourdoughs, one for each type of flour, were obtained by using selected LABs and yeast, previously characterized for different technological and nutritional features [5; 17; 18]. The sourdoughs were then used to produce the respective breads.