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Plant sterols (PSs) have reported benefits in alleviating colitis in mice, but the mechanisms involved require further investigation. This study aimed to evaluate the effect of a dietary supplement containing PSs (PS-DS) on inflammation biomarkers in a mice model of chronic ulcerative colitis induced by dextran sulfate sodium (DSS). C57BL/6J mice (n = 34) were exposed to 1.5% DSS in drinking water for three 5-day periods, with 10-day rest intervals in between. The mice received daily PS-DS (35 mg PS/kg) or placebo by oral gavage, either simultaneously (treatment) or 30 days prior (pre-treatment) to DSS exposition. After euthanasia, myeloperoxidase (MPO) activity and the levels of pro- (TNF-α and IL-6) and anti-inflammatory cytokines (IL-10) in the colonic tissue were analyzed. PS-DS treatment reduced (vs. DSS + placebo) myeloperoxidase (MPO) activity (2.6 ± 0.2 vs. 2.1 ± 0.1-fold change) and levels of TNF-α (85 ± 11 vs. 39 ± 7 pg/mg protein) and IL-6 (214 ± 26 vs. 128 ± 18 pg/mg protein), increasing the levels of IL-10 (46 ± 5 vs. 136 ± 16 pg/mg protein). PS-DS pre-treatment provided a greater inhibition (vs. treatment) of MPO activity (2.1 ± 0.3 vs. 1.3 ± 0.1-fold change) and a greater increase in IL-10 levels (50 ± 9 vs. 178 ± 30 pg/mg protein). These findings suggest that PS-DS has the potential to alleviate colitis in mice by modulating the inflammatory response and reducing oxidative stress. However, studies in humans are required to validate and fully understand its anti-inflammatory effect.

Studies on the impact of colonic fermentation of plant sterol (PS)-enriched foods using dynamic in vitro models are limited. This study aimed to evaluate the effect of a 72-h dynamic in vitro digestion-colonic fermentation (using the simgi® system) of PS-enriched rye bread on colonic microbial population and short-chain fatty acid (SCFA) and ammonium production. In all colon compartments (ascending colon (AC), transverse colon (TC), and descending colon (DC)) (72 vs. 0 h), a reduction in ammonium concentration (5.7- to 9.4-fold) and an increase in Staphylococcus spp. (1.4- to 2.1-fold), Lactobacillus spp. (1.5-fold), Bifidobacterium spp. (1.3- to 1.5-fold), and Enterococcus spp. (1.4- to 1.6-fold) were observed. In AC and TC, total SCFA decreased (4- and 1.3-fold, respectively), whereas it increased 1.4-fold in DC due to an increase in butyrate content from 4 to 45 mM. These results suggest that PS-enriched rye bread favors the growth of beneficial microbial species and the production of butyrate, a fuel source for enterocytes, thus promoting health benefits.

Rice, tiger nut and carob are Mediterranean products suitable for developing new foods, such as fermented beverages, due to their nutritional properties. These crops have a high carbohydrate content, are gluten and lactose-free and have a low allergenicity index. The development of fermented beverages from these crops can contribute to the Sustainable Development Goals by promoting human health and sustainable production and consumption. A narrative review of the nutritional value and potential functional activity of fermented beverages made from these crops was carried out. This literature review of existing studies on fermented and non-fermented beverages highlights their composition, production methodology, and health benefits. Fermented beverages made from these crops are high in fiber, essential fatty acids, vitamins (group B), and minerals. Fermentation increases the bioaccessibility of these nutrients while decreasing possible anti-nutritional factors. These fermented beverages offer several health benefits due to their antioxidant effects, modulating the intestinal microbiota and reducing the incidence of chronic degenerative diseases such as metabolic syndrome. Therefore, fermented rice, tiger nut and carob beverages can improve the Spanish diet by offering improved nutritional value and beneficial health effects. Additionally, these local crops promote sustainability, making them an appropriate choice for developing new fermented beverages.

Cell death program of red blood cells (RBCs), called eryptosis, is characterized by activation of caspases and scrambling of membrane phospholipids with externalization of phosphatidylserine (PS). Excessive eryptosis confers a procoagulant phenotype and is implicated in impairment of microcirculation and increased prothrombotic risk. It has recently been reported that cigarette smokers have high levels of circulating eryptotic erythrocytes, and a possible contribution of eryptosis to the vaso-occlusive complications associated to cigarette smoke has been postulated. In this study, we demonstrate how a mixture of plant sterols (MPtS) consisting of β-sitosterol, campesterol and stigmasterol, at serum concentration reached after ingestion of a drink enriched with plant sterols, inhibits eryptosis induced by cigarette smoke extract (CSE). Isolated RBCs were exposed for 4 h to CSE (10–20% v/v). When RBCs were co-treated with CSE in the presence of 22 µM MPtS, a significant reduction of the measured hallmarks of apoptotic death like assembly of the death-inducing signaling complex (DISC), PS outsourced, ceramide production, cleaved forms of caspase 8/caspase 3, and phosphorylated p38 MAPK, was evident. The new beneficial properties of plant sterols on CSE-induced eryptosis presented in this work open new perspectives to prevent the negative physio-pathological events caused by the eryptotic red blood cells circulating in smokers.

Bread is one of the staple foods of many countries, and its enrichment with bioactive compounds is trending. This phenomenon is focused on breads with a good nutritional profile, such as wholemeal rye bread (WRB), in which enrichment with plant sterols (PSs) is allowed in accordance with European regulations. The objective of the present study was to optimize the production of a WRB enriched with PS (PS-WRB) and to evaluate the proximate composition and starch digestibility as an indicator of nutritional quality. The rheological analysis showed that the bread dough presents satisfactory farinographic properties (dough development time 6 min; stability 4 min; degree of softening 100 Brabender units) but high water absorption (67%). The PS-WRB is high in dietary fiber and low in protein (20.4 and 7.7% w/w, dry basis, respectively) compared with other cereals reported in the scientific literature. In turn, a low starch proportion was hydrolyzed during the simulated digestion (59.9% of total starch), being also slowly hydrolyzed, as deduced from the rapidly digestible starch value (56.5% of total starch). In conclusion, WRB is a suitable matrix for PS enrichment, which allows for obtaining a product with a good nutritional profile and potential health benefits.

The fortification of foods with cholesterol-lowering agents such as plant sterols (PS) is allowed in Europe and could be of interest to the elderly since aging may lead to higher cardiovascular risk. To date, no studies have analyzed how the gastrointestinal conditions of the elderly affect PS bioaccessibility. Thus, this study evaluates the impact of adapting the standardized INFOGEST 2.0 method for adult to elderly physiological conditions on the bioaccessibility (BA) of PS-enriched wholemeal rye bread. For this purpose, changes in gastric or gastric and intestinal phase conditions (enzyme activity, pH, digestion time, or agitation) were made. Compared to adults, only when gastric and intestinal phase conditions were modified, the BA (%) decreased for individual (10.9–20.5 vs. 19.5–36.3) and total PSs (11.4 ± 1.7 vs. 20.5 ± 1.0), demonstrating that this reduction is due to intestinal phase modifications. However, the order of BA was not modified under any of the conditions tested, following the increasing rank: β-sitosterol, campesterol, Δ7-stigmasterol, sitostanol, Δ5,24, stigmastadienol, Δ5-avenasterol, Δ7avenasterol, stigmasterol, and campestanol. This complex matrix, very rich in fiber, such as wholemeal rye bread, may be responsible for the enzymatic reduction affecting PS solubility in the adapted digestion elderly model.

The effect of the incorporation of key enzymes of the lipidic metabolism to the INFOGEST (Improving health properties of food by sharing the knowledge on the digestive process) digestion method on plant sterol (PS) bioaccessibility is evaluated in a PS-enriched wholemeal rye bread. The assayed conditions were: (i) INFOGEST method; (ii) INFOGEST+gastric lipase (GL) (60 U/mL gastric digesta); and (iii) INFOGEST+GL+cholesterol esterase (CE) (0.075 U/mL intestinal digesta). The contents of PSs were determined in bioaccessible fractions after saponification and derivatization into trimethylsilylethers and were then analyzed by gas chromatography-mass spectrometry (for identification purposes) and gas chromatography-flame ionization detection (for quantification). The identified PSs from the PS-ingredient and rye flour were: campesterol, campestanol, stigmasterol, b-sitosterol, sitostanol, D5-avenasterol, D5,24-stigmastadienol, D7-stigmastenol and D7-avenasterol. The incorporation of the lipidic enzymes slightly reduced the bioaccessibility of the total (from 23.2 to 17.6–18.2%) and individual PS (from 22.6–57.7 to 17.3–44%). In addition, no differences in bioaccessibility were detected when GL or GL+CE were used. The largest bioaccessibility values were shown for D5-avenasterol and D7-avenasterol, regardless of the conditions assayed. In conclusion, the use of GL and CE means a closer approach to in vivo conditions, and we propose their inclusion in the INFOGEST model for the evaluation of the bioaccessibility of sterols and other lipid bioactive compounds.

In recent years, the food industry has increased its interest in the development of functional foods, including fruit juices, due to the increased demand among consumers for foods and beverages that benefit and improve our health [...]

Combining natural products as co-adjuvants in 5-fluorouracil (5-FU) chemotherapy might enhance the effectiveness of 5-FU by avoiding a high dosage and/or reducing treatment times. We explored the anticancer efficacy of the phytosterols (PS) at concentrations achievable in the human colon, as well as their potential as sensitizing agents of human colon cancer cells (Caco-2 and HT-29) to 5-FU treatment. Cells proliferation, combination index, cell cycle, apoptosis, caspases activation, ROS production, and ΔΨm were determined. Co-treatment (PS+5-FU) had an antiproliferative additive effect, and moreover, in general a significantly improved efficacy was observed on cell cycle arrest at S phase, apoptosis induction and increase in caspases activation compared to 5-FU alone, while no significant effects were observed on ROS levels and ΔΨm. Our results suggest, for the first time, the ability of PS to sensitize colon cancer cells to 5-FU chemotherapy and warrant further in vivo studies to confirm our preclinical findings.

Human red blood cells (RBCs), senescent or damaged due to particular stress, can be removed by programmed suicidal death, a process called eryptosis. There are various molecular mechanisms underlying eryptosis. The most frequent is the increase in the cytoplasmic concentration of Ca2+ ions, later exposure of erythrocytes to oxidative stress, hyperosmotic shock, ceramide formation, stimulation of caspases, and energy depletion. Phosphatidylserine (PS) exposed by eryptotic RBCs due to interaction with endothelial CXC-Motiv-Chemokin-16/Scavenger-receptor, causes the RBCs to adhere to vascular wall with consequent damage to the microcirculation. Eryptosis can be triggered by various xenobiotics and endogenous molecules, such as high cholesterol levels. The possible diseases associated with eryptosis are various, including anemia, chronic kidney disease, liver failure, diabetes, hypertension, heart failure, thrombosis, obesity, metabolic syndrome, arthritis, and lupus. This review addresses and collates the existing ex vivo and animal studies on the inhibition of eryptosis by food-derived phytochemicals and natural compounds including phenolic compounds (PC), alkaloids, and other substances that could be a therapeutic and/or co-adjuvant option in eryptotic-driven disorders, especially if they are introduced through the diet.

This study for the first time compares the in vivo (response in serum from individuals of an interventional study) and in vitro effects (bioaccessibility determined from simulated gastrointestinal digestion) of two plant sterol (PS)-enriched milk-based fruit beverages (with and without tangerine fruit juice rich in β-cryptoxanthin (β-Cx)) in order to assess their suitability as PS-enriched food matrixes, and the possible interactions between the two bioactive compounds (β-Cx and PS) that might affect their absorption. The randomized, double-blind, crossover study (performed in 36 postmenopausal women) showed the β-sitosterol and campesterol serum concentrations after four weeks of consumption of the beverages (1.5 g PS/day) to increase significantly (ranging 1.6–2.4 µg/mL and 0.3–0.4 µg/mL, respectively), thereby confirming suitability as matrices for PS-enrichment. The results therefore indicated similar behavior of PS supplied by these beverages and evaluated through in vitro or in vivo assays, independently of whether the samples contained β-Cx or not.

The INFOGEST digestion method was adapted for the evaluation of sterol bioaccessibility in a plant sterol (PS)-enriched beverage [1]. Recent modifications of the method, such as the addition of gastric lipase and cholesterol esterase, showed that the presence of sterols in digestion reagents has an important role on sterol bioaccessibility [2]. However, the specific sterol contribution from these extracts is unknown. Therefore, in the present study, the sterol content of the extracts used in the INFOGEST 2.0 digestion method (rabbit gastric extract—RGE, porcine pancreatin and bovine bile) is determined via GC-FID. The results show that the cholesterol content of the extracts is as follows: bovine bile > porcine pancreatin > RGE (4.07 ± 0.18, 1.41 ± 0.07 and 0.57 ± 0.05 mg/g extract, respectively). Considering the amount of each extract added to the digestion, the greatest cholesterol contributor is porcine pancreatin, followed by bovine bile and RGE (1.72 ± 0.08, 1.00 ± 0.04, and 0.046 ± 0.004 mg, respectively). However, cholesterol in bile is found in pre-formed micelles that increase its solubility, and, therefore, it would compete for sterol micellarization against sterol provided by the digested foods. This fact has been observed in previous studies in which the cholesterol content of the blanks of digestion was inversely correlated with PS bioaccessibility [2]. On the other hand, bovine bile contains stigmasterol, β-sitosterol and sitostanol (0.14 ± 0.01, 0.93 ± 0.07 and 0.18 ± 0.01 mg/g extract, respectively), while porcine pancreatin contains campesterol, stigmasterol, β-sitosterol and sitostanol (0.103 ± 0.004, 0.25 ± 0.02, 2.17 ± 0.42 and 0.32 ± 0.01 mg/g extract, respectively). Nevertheless, these PSs present in the extracts appear in negligible amounts in the digestion blank, reflecting their low solubility vs. cholesterol. In conclusion, in order to optimize the INFOGEST 2.0 gastrointestinal method for the evaluation of sterol bioaccessibility, the characterization of the sterol content in digestion reagents provides valuable information since it may affect their solubility.