José Manuel Silván

Nonextractable polyphenols, usually ignored, are the major part of dietary polyphenols: A study on the Spanish diet

SCOPE Dietary polyphenols (PP) can be divided into two groups: extractable polyphenols (EPP) or compounds solubilized by aqueous organic solvents, and nonextractable polyphenols (NEPP) or compounds that remain in their corresponding extraction residues. Most studies on food polyphenols and dietary intakes address exclusively EPP. The objective of this work was to determine the actual amount of PP, including NEPP, in food and in a whole diet. METHODS AND RESULTS HPLC-MS analyses were performed to identify EPP in methanol-acetone extracts and NEPP in the acidic hydrolyzates of their extraction residues in cereals, fruits, vegetables, nuts, and legumes. NEPP contents, estimated as hydrolyzable PP plus nonextractable proanthocyanidins (PA), ranged from 880 mg/100 g dry weight in fruits to 210 mg/100 g in cereals and were substantially higher than the contents of EPP. NEPP intake (day/person) in the Spanish diet (942 mg) is higher than EPP intake (258 mg) fruits and vegetables (746 mg) are the major contributors to the total PP intake (1201 mg). CONCLUSION Non extractable polyphenols are the major part of dietary polyphenols. The knowledge of intakes and physiological properties of NEPP may be useful for a better understanding of the potential health effects of dietary PP.

Protein Quality, Antigenicity, and Antioxidant Activity of Soy-Based Foodstuffs

Commercial soy-based foodstuffs, including beverages ( n = 15), cows milk supplemented with soy isoflavones ( n = 1), snacks ( n = 1), and biscuits ( n = 2), were analyzed to find any link between alterations in protein quality, safety (antigenicity), functionality (antioxidant activity), and food processing. Protein content was analyzed by the Kjeldhal method and available lysine by OPA assay. Chromatographic (RP-HPLC) and electrophoretic (SDS-PAGE) protein profiles were obtained to monitor modifications in the structure of soy allergens. The antigenicity was estimated by immunoblotting against soy total antibodies. Total phenol content was measured by Folin-Ciocalteu, while peroxyl radical scavenging activity of the sample was determined by ORAC FL assay. Protein content did not differ of those declared by the producers. Lysine availability was higher in liquid soy beverages compared to that in other soy foodstuffs studied here. 7S and 11S soy allergens were detected by RP-HPLC and SDS-PAGE, respectively. Both data indicated changes in soy protein patterns due to processing of instant powdered soymilk, soy snacks, and biscuits. Immunoblotting assay showed modifications in the antigenic response of these foodstuffs based on soy, suggesting that their processing had altered the structure of soy allergens. RP-HPLC, SDS-PAGE, and immunoblotting resulted in adequate analytical approaches for detecting changes in protein structure due to processing and adulteration. Protein quality, antigenicity, and antioxidant activity of soy products can be affected as a function of the intensity of the thermal processing.

Conceptual study on maillardized dietary fiber in coffee.

There is a methodological and conceptual overlap between coffee melanoidins and dietary fiber. Green Uganda coffee beans were roasted in a range from 8.1 to 21.6% of weight loss to evaluate melanoidins and dietary fiber. Samples were characterized by color, moisture, solubility, water activity, carbohydrates, polyphenols, protein, soluble dietary fiber (SDF), and melanoidins content. Hydroxymethylfurfural and chlorogenic acids were also measured as chemical markers of the extent of roasting. Melanoidins rapidly increased from 5.6 (light roasting) to 29.1 mg/100 mg soluble dry matter (dark roasting). A melanoidins-like structure was already present in green coffee that might overestimate up to 21.0% of the melanoidins content as determined by colorimetric methods. However, its contribution is variable and very likely depends on the method of drying applied to green coffee. SDF content (mg/100 mg soluble dry matter) gradually increased from 39.4 in green coffee to 64.9 at severe roasting conditions due to incorporation of neoformed colored structures and polyphenols. Then, SDF progressively turns to a maillardized structure, which increased from 11.0 to 45.0% according to the roasting conditions. It is concluded that the content of coffee melanoidins includes a substantial part of dietary fiber and also that coffee dietary fiber includes melanoidins. A conceptual discussion on a new definition of coffee melanoidins as a type of maillardized dietary fiber is conducted.

Peptides derived from in vitro gastrointestinal digestion of germinated soybean proteins inhibit human colon cancer cells proliferation and inflammation

The aim was to investigate the potential of germinated soybean proteins asa source of peptides with anticancer and anti-inflammatory activities produced after simulated gastrointestinal digestion. Protein concentrate from germinated soybean was hydrolysed with pepsin/pancreatin and fractionated by ultrafiltration. Whole digest and fractions>10, 5-10, and<5kDa caused cytotoxicity to Caco-2, HT-29, HCT-116 human colon cancer cells, and reduced inflammatory response caused by lipopolysaccharide in macrophages RAW 264.7. Antiproliferative and anti-inflammatory effects were generally higher in 5-10kDa fractions. This fraction was further purified by semi-preparative chromatography and characterised by HPLC-MS/MS. The most potent fraction was mainly composed of β-conglycinin and glycinin fragments rich in glutamine. This is the first report on the anti-cancer and anti-inflammatory effects of newly isolated and identified peptides from germinated soybean released during gastrointestinal digestion. These findings highlight the potential of germination as a process to obtain functional foods or nutraceuticals for colon cancer prevention.

Anthocyanins do not influence long-chain n-3 fatty acid status: Studies in cells, rodents and humans

Increased tissue status of the long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA), eicosapentaenoic (EPA) and docosahexaenoic acid (DHA) is associated with cardiovascular and cognitive benefits. Limited epidemiological and animal data suggest that flavonoids, and specifically anthocyanins, may increase EPA and DHA levels, potentially by increasing their synthesis from the shorter-chain n-3 PUFA, α-linolenic acid. Using complimentary cell, rodent and human studies we investigated the impact of anthocyanins and anthocyanin-rich foods/extracts on plasma and tissue EPA and DHA levels and on the expression of fatty acid desaturase 2 (FADS2), which represents the rate limiting enzymes in EPA and DHA synthesis. In experiment 1, rats were fed a standard diet containing either palm oil or rapeseed oil supplemented with pure anthocyanins for 8 weeks. Retrospective fatty acid analysis was conducted on plasma samples collected from a human randomized controlled trial where participants consumed an elderberry extract for 12 weeks (experiment 2). HepG2 cells were cultured with α-linolenic acid with or without select anthocyanins and their in vivo metabolites for 24 h and 48 h (experiment 3). The fatty acid composition of the cell membranes, plasma and liver tissues were analyzed by gas chromatography. Anthocyanins and anthocyanin-rich food intake had no significant impact on EPA or DHA status or FADS2 gene expression in any model system. These data indicate little impact of dietary anthocyanins on n-3 PUFA distribution and suggest that the increasingly recognized benefits of anthocyanins are unlikely to be the result of a beneficial impact on tissue fatty acid status.

Dietary gallic acid and anthocyanin cytotoxicity on human fibrosarcoma HT1080 cells. A study on the mode of action

Gallic acid and anthocyanins are abundant plant food bioactives present in many fruits and vegetables, being especially important in the composition of berries. Gallic acid has been shown to possess cytotoxic properties in several cancer cell lines and to inhibit carcinogenesis in animal models. However, its mechanism of action is not yet fully understood. The aim of this study was to elucidate whether the observed inhibitory activity of gallic acid against gelatinases corresponds to its cytotoxic activity in HT1080 cells and to determine if anthocyanins could exhibit a similar behavior. Gallic acid and delphinidin-3-glucoside have shown selective cytotoxicity towards HT1080 cells. Further analysis by a migration and invasion assay showed anti-invasive activities of gallic acid, delphinidin and pelargonidin-3-glucosides. Zymographic analysis demonstrated the inhibitory activity of gallic acid at the level of secreted and activated gelatinases. Moreover, gallic acid inhibited MMP-2 and MMP-9 proteolytic activity with very similar potency. NMR and molecular modelling experiments confirmed the interaction of gallic acid with MMP-2, and suggested that it takes place within the catalytic center. In this work we give some new experimental data supporting the role of these compounds in the inhibition of metalloproteases as the mechanism for their cytotoxic activity against fibrosarcoma.

Individual contributions of Savinase and Lactobacillus plantarum to lentil functionalization during alkaline pH-controlled fermentation

Legumes offer the possibility to develop multifunctional foods with benefits for metabolic syndrome. Our objective was to study the effect of alkaline fermentation by Lactobacillus plantarum and Savinase (FLPS) as well the individual effects of both processes on peptides, phenolics and bioactivity of lentil. FLPS increased peptides and some flavonoids and enhanced antioxidant activity, inhibition of angiotensin I-converting enzyme (ACE) and intestinal maltase activities of lentil soluble fraction. Savinase contributed to peptide release, ACE inhibitory and antioxidant activities of lentil soluble fraction. L. plantarum affected to phenolic composition, α-glucosidase and lipase inhibitory activities. Mass spectrometry analysis of the most active fermented lentil subfraction allowed the identification of the main bioactive compounds. Gastrointestinal digestion of fermented lentil increased bioaccessibility of peptides and phenolics as well as antioxidant activity. FLPS enhanced the overall healthy potential of lentil offering the possibility of its use as strategy for lentil functionalization.

pH-controlled fermentation in mild alkaline conditions enhances bioactive compounds and functional features of lentil to ameliorate metabolic disturbances

Lentil fermentation has a promising potential as a strategy for development of multifunctional ingredients targeting metabolic syndrome (MetS). Response surface methodology was applied to optimize lentil fermentation and study its effects on generation of peptides, soluble phenolics and bioactivities. Fermentation using Lactobacillus plantarum and Savinase® 16 L was carried out at different pH (6.5-8.5) and times (5.5-30 h). Analysis of variance was performed to evaluate linear, quadratic and interaction effects between fermentation parameters. pH positively affected peptides, soluble phenolic compounds and antioxidant activity whereas a negative impact on lipase inhibitory activity was observed (p < .0001). Time showed positive effect on proteolysis and negatively affected angiotensin I-converting enzyme inhibitory activity of fermented lentil (p < .0001). Multivariate optimization led to high levels of peptides, soluble phenolics and bioactivity of fermented lentil at pH 8.5 and 11.6 h. In conclusion, this study might contribute to the development of functional ingredients from lentil for MetS management.

Role of the polycarboxylic compounds in the response of Silene vulgaris to chromium

This work aims to investigate the nature and the specific mechanisms by which polycarboxylic compounds participate in the tolerance of Silene vulgaris to Cr with special attention given to the rhizosphere system. This knowledge is important to use this species in the implementation of phytoremediation technologies in Cr-polluted soils. According to the results, chromium is chelated and mobilized by the citric and malic acids in plant tissues, while oxalic acid might participate in the reduction and chelation of Cr in the rhizosphere. At the applied doses, the response of both exudation rate and root exudate composition (total polyphenols and quercitin) seems to involve a rearrangement in the lignification of the plant cell wall to immobilize Cr. Quercetin-3-dirhamnosyl-galactoside and apiin (apigenin-7-O-apiosyl-glucoside) have been identified as the major polyphenols in the root exudates of S. vulgaris. The increments found in the apiin concentration in root exudates seem to be related to the protection against Cr toxicity by chelation of Cr or by free radical scavenging. Though earlier response is detected in plant tissues, results from this work together with previous studies in S. vulgaris indicate that exudation might be a regulated mechanism of protection under Cr exposition in S. vulgaris that may involve mainly Cr reduction and chelation.

Effect of Long-Term Xanthophyll and Anthocyanin Supplementation on Lutein and Zeaxanthin Serum Concentrations and Macular Pigment Optical Density in Postmenopausal Women

Xanthophylls (lutein, L; zeaxanthin, Z) and anthocyanins are often included in food supplements to improve ocular health. There are no dietary reference intakes for them. The aim was to assess the effects of L, Z and anthocyanin supplementation on short and long-term lutein status markers (serum concentration and macular pigment optical density (MPOD)). Seventy-two postmenopausal women were randomized into a parallel study of 8 months: Group A—anthocyanines (60 mg/day); Group X—xanthophylls (6 mg L + 2 mg Z/day); Group X+A—anthocyanines (60 mg/day) + xanthophylls (6 mg L + 2 mg Z/day). At the beginning of the study, 4 and 8 month serum L and Z concentrations were determined (HPLC), as well as L, Z and anthocyanine dietary intake and MPOD (heterochromic flicker photometry). Baseline concentrations of L (0.35 ± 0.19 μmol/L), Z (0.11 ± 0.05 μmol/L), L+Z/cholesterol/triglycerides (0.07 ± 0.04 μmol/mmol) increased in Group X (2.8- and 1.6-fold in L and Z concentrations) and in group XA (2- and 1.4-fold in L and Z concentrations). MPOD (baseline: 0.32 ± 0.13 du) was not modified in any of the groups at the end of the study. There were no differences in the dietary intake of L+Z and anthocyanin at any point in time in any group. Supplementation of L and Z at a dietary level provoked an increase in their serum concentration that was not modified by simultaneous supplementation with anthocyanins.