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The aim of this work was to evaluate the influence of in vitro gastric digestion of two olive leaf extracts (E1 and E2) on their chemical composition and bioactive properties against Helicobacter pylori ( H. pylori ), one of the most successful and prevalent human pathogens. HPLC-PAD/MS analysis and anti-inflammatory, antioxidant, and antibacterial activities of both olive leaf extracts were carried out before and after their in vitro gastric digestion. The results showed that gastric digestion produced modifications of the chemical composition and bioactive properties of both olive leaf extracts. The main compounds in the extract E1 were hydroxytyrosol and its glucoside derivatives (14,556 mg/100 g), presenting all the identified compounds a more polar character than those found in the E2 extract. E2 showed a higher concentration of less polar compounds than E1 extract, with oleuropein (21,419 mg/100 g) being the major component. Gastric digestion during the fasted state (pH 2) induced an overall decrease of the most identified compounds. In the extract E1, while the anti-inflammatory capacity showed only a slight decrease (9% of IL-8 production), the antioxidant properties suffered a drastic drop (23% of ROS inhibition), as well as the antibacterial capacity. However, in the extract E2, these changes caused an increase in the anti-inflammatory (19% of IL-8 production) and antioxidant activity (9% of ROS inhibition), which could be due to the hydrolysis of oleuropein and ligustroside into their main degradation products, hydroxytyrosol and tyrosol, but the antibacterial activity was reduced. Gastric digestion during fed state (pH 5) had less influence on the composition of the extracts, affecting in a lesser degree their anti-inflammatory and antioxidant activity, although there was a decrease in the antibacterial activity in both extracts similar to that observed at pH 2.

Strains of Helicobacter pylori (H. pylori) resistant to various antibiotics have increased in recent years. In this context, the search for new therapeutic approaches is crucial. The aim of the present study was to demonstrate the antibacterial activity of a procyanidin-rich extract obtained from food-grade winery grape seeds against 14 H. pylori strains and elucidate its phenolic composition. Ten strains (71.4%) showed resistance to at least some of the tested antibiotics, while four isolates (28.6%) were susceptible to all antibiotics. Resistance to more than one class of antibiotics was observed in six strains (42.9%). The extract was able to inhibit the growth of all H. pylori strains in a range of a minimum inhibitory concentration (MIC) from 0.015 mg/mL to 0.125 mg/mL, confirming also the existence of a strain-dependent effect. The phenolic composition determined by reverse phase high pressure liquid chromatography, photodiode array, and mass spectrometry detection (RP-HPLC-PAD-MS) analysis revealed the presence of 43 individual compounds and allowed the quantification of 41 of them, including seven procyanidin tetramers, seven procyanidin pentamers, and six galloylated procyanidin dimers, trimers, and tetramers. The extract was composed mainly by catechin and procyanidin oligomers with a total amount of 5.801 mg/100 g, which represent 92% of the total individual phenolic content. Among them, the most abundant were catechins (2.047 mg/100 g), followed by procyanidin dimers (1.550 mg/100 g), trimers (1.176 mg/100 g), tetramers (436 mg/100 g), and pentamers (296 mg/100 g) that represent 35, 27, 20, 8, and 5%, respectively of the total flavanol constituents. The composition profile information may help to improve the production process of useful antibacterial extracts against H. pylori .

Pressurized liquid extraction with ethanol:water mixtures was proposed for obtaining phenolic antioxidants from grape stems. The optimal extraction conditions were elucidated by using a central composite rotatable design (solvent (X 1 , 0-100% ethanol:water v / v ), temperature (X 2 , 40-120 °C) and time (X 3 , 1-11 min)). Response surface methodology determined 30% ethanol:water, 120 °C and 10 min as the optimal extraction conditions regarding total phenolic content (TPC) (185.3 ± 2.9 mg gallic acid/g of extract) and antioxidant activity (3.55 ± 0.21 mmol Trolox/g, 1.22 ± 0.06 mmol Trolox/g and 1.48 ± 0.17 mmol Trolox/g of extract in ABTS, DPPH and ORAC methodologies, respectively). The antioxidant activity was attributed to total polymer procyanidins and flavan-3-ol monomers and oligomers, although other phenolic compound contributions should not be ruled out. Forty-two phenolic compounds were identified in the optimal extract, mainly polymer procyanidins and, to a lesser extent, monomers and oligomers of flavan-3-ols, quercetin-3- O -glucuronide, ε-viniferin, gallic and caftaric acid. Ethyl gallate, ellagic acid, protocatechuic aldehyde, delphinidin-7- O -glucoside and cyanidin-3- O -glucoside were reported for the first time in grape stem extracts. In conclusion, this study highlights the use of this winery side stream as a source of antioxidants within a sustainable food system.

Melanoidins present in coffee silverskin, the only by-product of the roasting process, are formed via the Maillard reaction. The exact structure, biological properties, and mechanism of action of coffee silverskin melanoidins, remain unknown. This research work aimed to contribute to this novel knowledge. To achieve this goal, melanoidins were obtained from an aqueous extract of Arabica coffee silverskin (WO2013004873A1) and was isolated through ultrafiltration (>10 kDa). The isolation protocol was optimized and the chemical composition of the high molecular weight fraction (>10 kDa) was evaluated, by analyzing the content of protein, caffeine, chlorogenic acid, and the total dietary fiber. In addition, the structural analysis was performed by infrared spectroscopy. Antioxidant properties were studied in vitro and the fiber effect was studied in vivo, in healthy male Wistar rats. Melanoidins were administered to animals in the drinking water at a dose of 1 g/kg. At the fourth week of treatment, gastrointestinal motility was evaluated through non-invasive radiographic means. In conclusion, the isolation process was effective in obtaining a high molecular weight fraction, composed mainly of dietary fiber, including melanoidins, with in vitro antioxidant capacity and in vivo dietary fiber effects., Competing Interests: The authors declare no conflict of interest.