Toxic or beneficial? What is the role of food-relevant selenium species selenoneine?

Selenium (Se) is an essential trace element that is ubiquitously present in the environment in small concentrations. Essential functions of Se in the human body are manifested through the wide range of proteins, containing selenocysteine as their active center. Such proteins are called selenoproteins which are found in multiple physiological processes like antioxidative defense and the regulation of thyroid hormone functions. Therefore, Se deficiency is known to cause a broad spectrum of physiological impairments, especially in endemic regions with low Se content. Nevertheless, being an essential trace element, Se could exhibit toxic effects, if its intake exceeds tolerable levels. Accordingly, this range between deficiency and overexposure represents optimal Se supply. However, this range was found to be narrower than for any other essential trace element. Together with significantly varying Se concentrations in soil and the presence of specific bioaccumulation factors, this represents a noticeable difficulty in the assessment of Se epidemiological status. While Se is acting in the body through multiple selenoproteins, its intake occurs mainly in form of small organic or inorganic molecular mass species. Thus, Se exposure not only depends on daily intake but also on the respective chemical form, in which it is present. The essential functions of selenium have been known for a long time and its primary forms in different food sources have been described. Nevertheless, analytical capabilities for a comprehensive investigation of Se species and their derivatives have been introduced only in the last decades. A new Se compound was identified in 2010 in the blood and tissues of bluefin tuna. It was called selenoneine (SeN) since it is an isologue of naturally occurring antioxidant ergothioneine (ET), where Se replaces sulfur. In the following years, SeN was identified in a number of edible fish species and attracted attention as a new dietary Se source and potentially strong antioxidant. Studies in populations whose diet largely relies on fish revealed that SeN represents the main non-protein bound Se pool in their blood. First studies, conducted with enriched fish extracts, already demonstrated the high antioxidative potential of SeN and its possible function in the detoxification of methylmercury in fish. Cell culture studies demonstrated, that SeN can utilize the same transporter as ergothioneine, and SeN metabolite was found in human urine. Until recently, studies on SeN properties were severely limited due to the lack of ways to obtain the pure compound. As a predisposition to this work was firstly a successful approach to SeN synthesis in the University of Graz, utilizing genetically modified yeasts. In the current study, by use of HepG2 liver carcinoma cells, it was demonstrated, that SeN does not cause toxic effects up to 100 M concentration in hepatocytes. Uptake experiments showed that SeN is not bioavailable to the used liver cells. In the next part a blood-brain barrier (BBB) model, based on capillary endothelial cells from the porcine brain, was used to describe the possible transfer of SeN into the central nervous system (CNS). The assessment of toxicity markers in these endothelial cells and monitoring of barrier conditions during transfer experiments demonstrated the absence of toxic effects from SeN on the BBB endothelium up to 100 M concentration. Transfer data for SeN showed slow but substantial transfer. A statistically significant increase was observed after 48 hours following SeN incubation from the blood-facing side of the barrier. However, an increase in Se content was clearly visible already after 6 hours of incubation with 1 M of SeN. While the transfer rate of SeN after application of 0.1 M dose was very close to that for 1 M, incubation with 10 M of SeN resulted in a significantly decreased transfer rate. Double-sided application of SeN caused no side-specific transfer of SeN, thus suggesting a passive diffusion mechanism of SeN across the BBB. This data is in accordance with animal studies, where ET accumulation was observed in the rat brain, even though rat BBB does not have the primary ET transporter OCTN1. Investigation of capillary endothelial cell monolayers after incubation with SeN and reference selenium compounds showed no significant increase of intracellular selenium concentration. Speciesspecific Se measurements in medium samples from apical and basolateral compartments, as good as in cell lysates, showed no SeN metabolization. Therefore, it can be concluded that SeN may reach the brain without significant transformation. As the third part of this work, the assessment of SeN antioxidant properties was performed in Caco-2 human colorectal adenocarcinoma cells. Previous studies demonstrated that the intestinal epithelium is able to actively transport SeN from the intestinal lumen to the blood side and accumulate SeN. Further investigation within current work showed a much higher antioxidant potential of SeN compared to ET. The radical scavenging activity after incubation with SeN was close to the one observed for selenite and selenomethionine. However, the SeN effect on the viability of intestinal cells under oxidative conditions was close to the one caused by ET. To answer the question if SeN is able to be used as a dietary Se source and induce the activity of selenoproteins, the activity of glutathione peroxidase (GPx) and the secretion of selenoprotein P (SelenoP) were measured in Caco-2 cells, additionally. As expected, reference selenium compounds selenite and selenomethionine caused efficient induction of GPx activity. In contrast to those SeN had no effect on GPx activity. To examine the possibility of SeN being embedded into the selenoproteome, SelenoP was measured in a culture medium. Even though Caco-2 cells effectively take up SeN in quantities much higher than selenite or selenomethionine, no secretion of SelenoP was observed after SeN incubation. Summarizing, we can conclude that SeN can hardly serve as a Se source for selenoprotein synthesis. However, SeN exhibit strong antioxidative properties, which appear when sulfur in ET is exchanged by Se. Therefore, SeN is of particular interest for research not as part of Se metabolism, but important endemic dietary antioxidant.     

Bioavailability Assessment of Copper,Iron, Manganese,Molybdenum, Selenium,and Zinc from Selenium‐Enriched Lettuce

Cu, Fe, Mn, Mo, Selenium (Se), and Zn bioavailability from selenate‐ and selenite‐enriched lettuce plants was studied by in vitro gastrointestinal digestion followed by an assay with Caco‐2 cells. The plants were cultivated in the absence and presence of two concentrations (25 and 40 µmol/L of Se). After 28 days of cultivation, the plants were harvested, dried, and evaluated regarding the total concentration, bioaccessibility, and bioavailability of the analytes. The results showed that biofortification with selenate leads to higher Se absorption by the plant than biofortification with selenite. For the other nutrients, Mo showed high accumulation in the plants of selenate assays, and the presence of any Se species led to a reduction of the plant uptake of Cu and Fe. The accumulation of Zn and Mn was not strongly influenced by the presence of any Se species. The bioaccessibility values were approximately 71%, 10%, 52%, 84%, 71%, and 86% for Cu, Fe, Mn, Mo, Se, and Zn, respectively, and the contribution of the biofortified lettuce to the ingestion of these minerals is very small (except for Se and Mo). Due to the low concentrations of elements from digested plants, it was not possible to estimate the bioavailability for some elements, and for Mo and Zn, the values are below 6.9% and 3.4% of the total concentration, respectively. For Se, the bioavailability was greater for selenite‐enriched than selenate‐enriched plants (22% and 6.0%, respectively), because selenite is biotransformed by the plant to organic forms that are better assimilated by the cells.     

Apple extract induces increased epithelial resistance and claudin 4 expression in Caco-2 cells

BACKGROUND: The small intestinal epithelium functions both to absorb nutrients, and to provide a barrier between the outside, luminal, world and the human body. One of the passageways across the intestinal epithelium is paracellular diffusion, which is controlled by the properties of tight junction complexes. We used a differentiated Caco‐2 monolayer as a model for small intestinal epithelium to study the effect of crude apple extracts on paracellular permeability. RESULTS: Exposure of crude apple homogenate to the differentiated Caco‐2 cells increased the paracellular resistance, determined as trans‐epithelial electrical resistance (TEER). This increase was linearly related to the concentration of apple present. The TEER‐enhancing effect of apple extract was due to factors mainly present in the cortex, and the induction was not inhibited by protein kinase inhibitors. Apple‐induced resistance was accompanied by increased expression of several tight junction related genes, including claudin 4 (CLDN4). CONCLUSION: Crude apple extract induces a higher paracellular resistance in differentiated Caco‐2 cells. Future research will determine whether these results can be extrapolated to human small intestinal epithelia. Copyright © 2011 Society of Chemical Industry     

Physiological function of Se‐enriched tea fertilised with sodium selenite and naturally high‐Se tea in rats

In order to increase food selenium (Se) content, Se‐enriched tea was produced by fertilising with sodium selenite in low‐Se soil. Five groups of rats were fed a low‐Se diet supplemented with either water (Se‐deficient), sodium selenite or an aqueous extraction of low‐Se tea, Se‐enriched tea or naturally high‐Se tea. The chemical form of Se in Se‐enriched tea and the physiological function in rats fed the different Se sources were determined after 8 weeks. The results showed that organic Se accounted for 80% or more of the Se in Se‐enriched tea fertilised with sodium selenite and naturally high‐Se tea, but no significant differences in the proportion of organic Se and protein Se were found between Se‐enriched tea and naturally high‐Se tea. The Se biological utilisation rates were 65.41, 68.05 and 70.49% for sodium selenite, Se‐enriched tea and naturally high‐Se tea respectively. The Se content of blood and liver and the glutathione peroxidase (GSH‐Px) activity were significantly increased by feeding Se‐enriched tea and sodium selenite compared with low‐Se tea, but a more efficient increase in liver GSH‐Px activity was obtained with Se‐enriched tea than with sodium selenite. No significant differences were found between Se‐enriched tea fertilised with sodium selenite and naturally high‐Se tea, which proved that the biological effectiveness of Se in Se‐enriched tea was higher than that of sodium selenite in increasing liver GSH‐Px activity. Se‐enriched tea fertilised with sodium selenite in low‐Se soil gave the same biological function as naturally high‐Se tea. Therefore Se‐enriched tea is a safe and effective means of increasing the Se intake of both humans and animals in low‐Se areas. © 2000 Society of Chemical Industry     

Absorption and metabolism of the mycotoxin zearalenone and the growth promotor zeranol in Caco-2 cells in vitro

Scope: Zearalenone (ZEN) and α‐zearalanol (α‐ZAL, zeranol) were studied in differentiated Caco‐2 cells and in the Caco‐2 Millicell^® system in vitro to simulate their in vivo intestinal absorption and metabolism in humans. Methods and results: In addition to metabolic reduction/oxidation, extensive conjugation with glucuronic acid and sulfate of the parent compounds and their phase I metabolites was observed. The positional isomers of the glucuronides and sulfates were unambiguously identified: Sulfonation occurred specifically at the 14‐hydroxyl group, whereas glucuronidation was less specific and, in addition to the preferred 14‐hydroxyl group, involved the 16‐ and 7‐hydroxyl groups. Using the Caco‐2 Millicell^® system, an efficient transfer of the glucuronides and sulfates of ZEN and α‐ZAL and their phase I metabolites into both the basolateral and the apical compartment was observed after apical administration. The apparent permeability coefficients (P_app values) of ZEN, α‐ZAL and the ZEN metabolite α‐zearalenol were determined, using an initial apical concentration of 20 μM and a permeation time of 1 h. Conclusion: According to the P_app values, the three compounds are expected to be extensively and rapidly absorbed from the intestinal lumen in vivo and reach the portal blood both as aglycones and as glucuronide and sulfate conjugates in humans.     

Ginger Extract Suppresses Inflammatory Response and Maintains Barrier Function in Human Colonic Epithelial Caco‐2 Cells Exposed to Inflammatory Mediators

The beneficial effects of ginger in the management of gastrointestinal disturbances have been reported. In this study, the anti‐inflammatory potential of ginger extract was assessed in a cellular model of gut inflammation. In addition, the effects of ginger extract and its major active compounds on intestinal barrier function were evaluated. The response of Caco‐2 cells following exposure to a mixture of inflammatory mediators [interleukin [IL]‐1β, 25 ng/mL; lipopolysaccharides [LPS], 10 ng/mL; tumor necrosis factor [TNF]‐α, 50 ng/mL; and interferon [INF]‐γ, 50 ng/mL] were assessed by measuring the levels of secreted IL‐6 and IL‐8. In addition, the mRNA levels of cyclooxygenase‐2 and inducible nitric oxide synthase were measured. Moreover, the degree of nuclear factor (NF)‐κB inhibition was examined, and the intestinal barrier function was determined by measuring the transepithelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC)‐dextran transfer. It was observed that ginger extract and its constituents improved inflammatory responses by decreasing the levels of nitrite, PGE2, IL‐6, and IL‐8 via NF‐κB inhibition. The ginger extract also increased the TEER and decreased the transfer of FITC‐dextran from the apical side of the epithelium to the basolateral side. Taken together, these results show that ginger extract may be developed as a functional food for the maintenance of gastrointestinal health.     

The (193–209) 17‐residues peptide of bovine β‐casein is transported through Caco‐2 monolayer

Although the bioavailability of large peptides with biological activity is of great interest, the intestinal transport has been described for peptides up to only nine residues. β‐casein (β‐CN, 193–209) is a long and hydrophobic peptide composed of 17 amino acid residues (molecular mass 1881 Da) with immunomodulatory activity. The present work examined the transport of the β‐CN (193–209) peptide across Caco‐2 cell monolayer. In addition, we evaluated the possible routes of the β‐CN (193–209) peptide transport, using selective inhibitors of the different routes for peptide transfer through the intestinal barrier. The results showed that the β‐CN (193–209) peptide resisted the action of brush‐border membrane peptidases, and that it was transported through the Caco‐2 cell monolayer. The main route involved in transepithelial transport of the β‐CN (193–209) peptide was transcytosis via internalized vesicles, although the paracellular transport via tight‐junctions could not be excluded. Our results demonstrated the transport of an intact long‐chain bioactive peptide in an in vitro model of intestinal epithelium, as an important step to prove the evidence for bioavailability of this peptide.     

Bioactive dietary polyphenols inhibit heme iron absorption in a dose-dependent manner in human intestinal Caco-2 cells

Abstract: Although heme iron is an important form of dietary iron, its intestinal absorption mechanism remains elusive. Our previous study revealed that (‐)‐epigallocatechin‐3‐gallate (EGCG) and grape seed extract (GSE) markedly inhibited intestinal heme iron absorption by reducing the basolateral iron export in Caco‐2 cells. The aim of this study was to examine whether small amounts of EGCG, GSE, and green tea extract (GT) could inhibit heme iron absorption, and to test whether the inhibitory action of polyphenols could be offset by ascorbic acid. A heme‐⁵⁵Fe absorption study was conducted by adding various concentrations of EGCG, GSE, and GT to Caco‐2 cells in the absence and presence of ascorbic acid. Polyphenolic compounds significantly inhibited heme‐⁵⁵Fe absorption in a dose‐dependent manner. The addition of ascorbic acid did not modulate the inhibitory effect of dietary polyphenols on heme iron absorption when the cells were treated with polyphenols at a concentration of 46 mg/L. However, ascorbic acid was able to offset or reverse the inhibitory effects of polyphenolic compounds when lower concentrations of polyphenols were added (≤ 4.6 mg/L). Ascorbic acid modulated the heme iron absorption without changing the apical heme uptake, the expression of the proteins involved in heme metabolism and basolateral iron transport, and heme oxygenase activity, indicating that ascorbic acid may enhance heme iron absorption by modulating the intracellular distribution of ⁵⁵Fe. These results imply that the regular consumption of dietary ascorbic acid can easily counteract the inhibitory effects of low concentrations of dietary polyphenols on heme iron absorption but cannot counteract the inhibitory actions of high concentrations of polyphenols. Practical Application: Bioactive dietary polyphenols inhibit heme iron absorption in a dose‐dependent manner. The small amounts of polyphenolic compounds present in foods are capable of reducing heme iron transport across the intestinal enterocyte. However, the inhibitory effects of dietary polyphenolic compounds on heme iron absorption can be offset by ascorbic acid and can possibly be avoided by decreasing the consumption of polyphenols while simultaneously taking ascorbic acid.     

Vitamin D intestinal absorption is not a simple passive diffusion: evidences for involvement of cholesterol transporters

Scope : It is assumed that vitamin D is absorbed by passive diffusion. However, since cholecalciferol (vitamin D₃) and cholesterol display similar structures, we hypothesized that common absorption pathways may exist. Methods and results : Cholecalciferol apical transport was first examined in human Caco‐2 and transfected Human embryonic kidney (HEK) cells. Cholecalciferol uptake was then valuated ex vivo and in vivo, using either wild‐type mice, mice overexpressing Scavenger Receptor class B type I (SR‐BI) at the intestinal level or mice treated or not with ezetimibe. Cholecalciferol uptake was concentration‐, temperature‐ and direction‐dependent, and was significantly impaired by a co‐incubation with cholesterol or tocopherol in Caco‐2 cells. Moreover Block Lipid Transport‐1 (SR‐BI inhibitor) and ezetimibe glucuronide (Niemann‐Pick C1 Like 1 inhibitor) significantly decreased cholecalciferol transport. Transfection of HEK cells with SR‐BI, Cluster Determinant 36 and Niemann‐Pick C1 Like 1 significantly enhanced vitamin D uptake, which was significantly decreased by the addition of Block Lipid Transport‐1, sulfo‐N‐succinimidyl oleate (Cluster Determinant 36 inhibitor) or ezetimibe glucuronide, respectively. Similar results were obtained in mouse intestinal explants. In vivo, cholecalciferol uptake in proximal intestinal fragments was 60% higher in mice overexpressing SR‐BI than in wild‐type mice (p<0.05), while ezetimibe effect remained non‐significant. Conclusion: These data show for the first time that vitamin D intestinal absorption is not passive only but involves, at least partly, some cholesterol transporters.     

Modified Dietary Fiber from Cassava Pulp and Assessment of Mercury Bioaccessibility and Intestinal Uptake Using an In Vitro Digestion/Caco‐2 Model System

The ability of modified dietary fiber (MDF) generated from cassava pulp to modulate the bioaccessibility and intestinal absorption of heavy metals may be helpful to mitigate health risk associated with select foods including select fish high in methyl mercury. Using a coupled in vitro digestion/Caco‐2 human intestinal cell model, the reduction of fish mercury bioaccessibility and intestinal uptake by MDF was investiaged. MDF was prepared from cassava pulp, a byproduct of tapioca production. The highest yield (79.68%) of MDF was obtained by enzymatic digestion with 0.1% α‐amylase (w/v), 0.1% amyloglucosidase (v/v) and 1% neutrase (v/v). MDF and fish tissue were subjected to in vitro digestion and results suggest that MDF may reduce mercury bioaccessibility from fish to 34% to 85% compared to control in a dose‐dependent manner. Additionally, accumulation of mercury from digesta containing fish and MDF was only modestly impacted by the presence of MDF. In conclusion, MDF prepared from cassava pulp may be useful as an ingredient to reduce mercury bioavailability from food such as fish specifically by inhibiting mercury transfer to the bioaccessibile fraction during digestion.     

Se in Se‐enriched peanut,and losses during peanut protein preparation

This study determined the Se species in Se‐enriched peanut, and Se losses during peanut protein processing by enzymatic extraction, high‐performance liquid chromatography (HPLC) separation and inductively coupled plasma‐mass spectrometry determination. The study revealed that mixed enzymes (protease and lipase, 2:1 w/w) in Na₂S₂O₃, assisted by 1 h ultrasonic processing, could effectively extract Se speciation from defatted peanut powder. Separation of organic Se by HPLC was optimised using pentafluoropropionic anhydride at a concentration of 0.1% in 2% methanol as mobile phase. Selenomethionine is the dominant Se species in peanut, accounting for 65% of the total Se. During the peanut protein preparation, nearly 37% of Se losses were due to the complexity of the multistage process. The loss can be ascribed to volatilisation, dissolution, degradation or other physical modes of transfer or loss.     

Absorption of anthocyanins through intestinal epithelial cells – Putative involvement of GLUT2

Anthocyanins bioavailability is a major issue regarding their biological effects and remains unclear due to few data available on this matter. This work aimed to evaluate the absorption of anthocyanins at the intestine using Caco‐2 cells. Anthocyanin extract, rich in malvidin‐3‐glucoside, was obtained from red grape skins and tested on Caco‐2 cells. The absorption of anthocyanins, in absence or presence of 1% ethanol, was detected by HPLC/DAD/LC‐MS. Our results showed that this transport was significantly increased in the presence of ethanol especially after 60 min of incubation. In addition, cells that were pretreated for 96 h with anthocyanins (200 μg/mL) showed an increase of their own transport (about 50% increase). Expression of glucose transporters sodium‐dependent glucose transporter 1, facilitative glucose transporters 5, and facilitative glucose transporters 2 was assessed by RT‐PCR. It was found that facilitative glucose transporters 2 expression was increased (60%) in Caco‐2 cells pretreated with anthocyanins, by comparison with controls. When the effect of anthocyanin extract on ³H‐2‐deoxy‐D ‐glucose uptake was tested, an inhibitory effect was observed (about 60% decrease). However, the malvidin aglycone was tested and had no effect. In conclusion, anthocyanins could be absorbed through Caco‐2 cells, and can interfere with their own transport and also with glucose intestinal uptake.     

Determination of selenium concentration in rice and the effect of foliar application of Se‐enriched fertiliser or sodium selenite on the selenium content of rice

Atomic fluorescence spectrophotometry was used to determine the selenium (Se) concentration in 30 rice products of different species from Southern China. The Se level ranged from 0.015 to 0.046 µg g^?1. Considering the average daily individual consumption of rice products in Southern China, the average dietary intake of Se supplied by this source is only 6–18 µg day^?1 for an adult. This low Se intake from rice products is mainly responsible for the low total Se intake of inhabitants in Southern China. Foliar spraying of Se‐enriched fertiliser or sodium selenite with 14–18 g Se ha^?1 in the heading stage of rice growth increased the Se content to 0.178–0.421 µg g^?1 in rice products. The concentration and amount of Se‐enriched fertiliser can be varied to achieve an optimum concentration of Se in rice products. The authors suggest that these Se‐enriched rice products can contribute an increase in Se intake of 50–100 µg day^?1 on average if their Se concentration is controlled in the region of 0.15–0.50 µg g^?1. © 2002 Society of Chemical Industry     

Effects of selenium addition on minimally processed leafy vegetables grown in a floating system

BACKGROUND: In recent years the consumption of minimally processed leafy vegetables has been increasing. At the same time food quality and its effects on human health have become crucial issues. Since selenium (Se) is an important microelement due to its ability to defend human organisms against free radicals, we investigated the effects of Se on chicory and lettuce production, and assessed the feasibility of a floating system as a method of producing Se‐enriched vegetables. RESULTS: The addition of 0.5 and 1.0 mg Se L^?1 to the nutrient solution resulted in an increase in the selenium concentration in the leaves, which had a positive effect on the plant yield. Selenium was generally effective in decreasing the production of ethylene and phenylalanine ammonia lyase (PAL) activity, even though a seasonal effect seemed to be present, consequently improving the quality of leafy vegetables and the shelf life in both species. The amount of Se accumulated in plants grown in the nutrient solution containing 0.5 mg Se kg^?1 could provide the rational Se intake for human nutrition in accordance with the recommended dietary allowance (RDA) guidelines. CONCLUSIONS: The addition of Se in a nutrient solution can be a useful system for providing enriched leafy vegetables. The floating system can be used to modulate the availability of Se in nutrient solutions and to grow vegetables with the optimal Se content for human health. Copyright © 2009 Society of Chemical Industry     

Effects of selenium form on blood and milk selenium concentrations,milk component and milk fatty acid composition in dairy cows

BACKGROUND: Human health may be improved if milk with a favorable fatty acid composition and Se concentration is ingested. The present study is to determine how a basal diet supplemented with daily 5 mg Se as Se‐enriched yeast (SY) or sodium selenite (SS) affects the fatty acid composition and Se concentration of bovine milk. The effects of Se form on blood Se concentration, erythrocyte glutathione peroxidase 1 (GPx1) activity, serum GPx3 activity and milk yield and component were also studied. RESULTS: Both Se forms, when compared to control group, increased Se concentrations of blood (P < 0.01) and milk (P < 0.01), erythrocyte GPx1 activity (P < 0.05) and milk percentages of polyunsaturated fatty acids (PUFA) (P < 0.05) and cis‐9,cis‐12 linoleic acid (P < 0.05). Cows supplemented with SY had higher Se levels in blood (P < 0.01) and milk (P < 0.01) and percentage of PUFA in milk (P < 0.05) when compared with those supplemented with SS. Milk yield, milk component and serum GPx3 activity were not significantly affected by Se form. CONCLUSION: Supplementation of diet with SY appears to be of more benefit than SS in producing favorable milk with high PUFA and Se concentrations. Copyright © 2010 Society of Chemical Industry     

Genomic Study of the Absorption Mechanism of p‐Coumaric Acid and Caffeic Acid of Extract of Ananas Comosus L. Leaves

Cardiac disease has emerged as the leading cause of death worldwide, and food rich in phenolic acids has drawn much attention as sources of active substances of hypolipidemic drug. Ananas comosus L. (pineapple) is one of the most popular tropical and subtropical fruits. Isolated from pineapple leaves, EAL(Extract of Ananas Comosus L. Leaves) is rich in phenolic acids, such as p‐coumaric acid, caffeic acid, and other phenolics, highly relevant to the putative cardiovascular‐protective effects, which suggests its potential to be a new plant medicine for treatment of cardiac disease, but little is known about absorption, distribution, metabolism, and excretion of EAL in animals or human beings. In this study, we employed cDNA microarray, Caco‐2 cell lines, and rat intestinal model to explore the absorption behavior of p‐coumaric acid and caffeic acid in EAL. The permeation of 2 substances was concentration and time dependent. Results also indicated that monocarboxylic acid transporter was involved in the transepithelial transport of p‐coumaric acid and caffeic acid.     

Dietary Amino Acids and the Gut‐Microbiome‐Immune Axis: Physiological Metabolism and Therapeutic Prospects

Dietary amino acids (AAs) are not only absorbed and metabolized by enterocytes but also available to the microbiota in the gut in mammals. In addition to serving as the materials for protein synthesis, AAs can act as precursors for numerous metabolic end products in reactions involving the intestinal mucosa and microbiota. After penetrating the epithelial barrier, microbial metabolites can enter and accumulate in the host circulatory system, where they are sensed by immune cells and then elicit a wide range of biological functions via different receptors and mechanisms. Some intestinal bacteria can also synthesize certain AAs, implying that the exchange of AAs between hosts and microorganisms is bidirectional. Changes in AA composition and abundance can affect AA‐metabolizing bacterial communities and modulate macrophages and dendritic cells via toll‐like receptors (TLRs), autoinducer‐2 (AI‐2), and NOD‐like receptors (NLRs), and also regulate the gut‐microbiome‐immune axis via aryl hydrocarbon receptor (AhR), serotonin/5‐hydroxytryptamine (5‐HT), and other signaling pathways, all of which play critical roles in regulating the intestinal mucosal immunity and microbiota directly or indirectly, contributing to intestinal homeostasis. Therefore, the current findings of the effects of certain functional AAs on the gut‐microbiome‐immune axis are reviewed, illustrating signaling pathways of tryptophan (Trp), glutamine (Gln), methionine (Met), and branched‐chain AAs (BCAAs) in the intestinal barrier and regarding immunity via crosstalk with their receptors or ligands. These findings have shed light on the clinical applications of dietary AAs in improving gut microbiota and mucosal immunity, therefore benefiting the gut as well as local and systemic health.     

Partially Hydrolyzed Soy Protein Shows Enhanced Transport of Amino Acids Compared to Nonhydrolyzed Protein across an Intestinal Epithelial Cell Monolayer

Consumption of protein hydrolysates has been proposed to stimulate muscle anabolism more than intact (nonhydrolyzed) proteins via accelerated delivery of amino acids for muscle protein synthesis (MPS). We evaluated whether the rate of amino acid uptake and transport across intestinal cells was enhanced for soy protein hydrolysates versus nonhydrolyzed soy protein. Intact and partially hydrolyzed proteins were subject to simulated gut digestion and applied to the apical surface of Caco‐2 monolayers. Basolateral media was harvested after 3 h and quantitatively analyzed for free amino acids using ion‐exchange chromatography and comparison to an included reference standard. Basolateral concentrations of all amino acids were higher (mean 32%) for hydrolyzed versus nonhydrolyzed protein with the greatest differences in histidine, lysine, and valine. Scale‐up production of the soy protein hydrolysate did not diminish its enhanced absorption properties. These data support the hypothesis that hydrolyzed soy protein may provide dietary amino acids that are more rapidly transported across the intestinal epithelium versus intact soy protein. This would be important under conditions where rapid and increased levels of amino acids are needed such as in the stimulation of MPS.     

Combined speciation analysis by X-ray absorption near-edge structure spectroscopy, ion chromatography, and solid-phase microextraction gas chromatography-mass spectrometry to evaluate biotreatment of concentrated selenium wastewaters

In this study we evaluate the potential of anaerobic granular sludge as an inoculum for the bioremediation of selenium-contaminated waters using species-specific analytical methods. Solid species formed by microbial reduction were investigated using X-ray absorption near-edge structure (XANES) spectroscopy at the selenium K-edge. Furthermore, dissolved selenium species were specifically determined by ion chromatography (IC) and solid-phase microextraction gas chromatography-mass spectrometry (SPME-GC-MS). Least-squares linear combination of the XANES spectra for samples incubated with the highest selenate/selenite concentrations (10(-3) M) show the predominance of elemental selenium and a Se(-I) selenide, such as ferroselite, the thermodynamically most stable iron selenide. In contrast, elemental selenium and Se(-II) selenides are the main species detected at the lower selenate/selenite concentrations. In each repeated fed batch incubation, most aqueous selenite anions were converted into solid selenium species, regardless of the type of electron donor used (acetate or H(2)/CO(2)) and the selenium concentration applied. On the other hand, at higher concentrations of selenate (10(-4) and 10(-3) M), significant amounts of the oxyanion remained unconverted after consecutive incubations. SPME-GC-MS demonstrated selenium alkylation with both electron donors investigated, as dimethyl selenide (DMSe) and dimethyl diselenide (DMDSe). Selenite was even more alkylated in the presence of H(2)/CO(2) (maximum 2156 μg of Se/L of DMSe + DMDSe) as compared to acetate (maximum 50 μg of Se/L). In contrast, selenate was less alkylated using both electron donors (maximum 166 and 3 μg of Se/L, respectively). The high alkylation potential for selenite limits its bioremediation in selenium laden waters involving H(2)/CO(2) as the electron donor despite the fact that nontoxic elemental selenium and thermodynamically stable metal selenide species are formed.     

Determination of S‐methyl‐L‐methionine (SMM) from Brassicaceae Family Vegetables and Characterization of the Intestinal Transport of SMM by Caco‐2 Cells

The objectives of the current study were to determine S‐methyl‐L‐methionine (SMM) from various Brassicaceae family vegetables by using validated analytical method and to characterize the intestinal transport mechanism of SMM by the Caco‐2 cells. The SMM is well known to provide therapeutic activity in peptic ulcers. The amount of SMM from various Brassicaceae family vegetables ranged from 89.08 ± 1.68 μg/g to 535.98 ± 4.85 μg/g of dry weight by using validated ultra‐performance liquid chromatography‐electrospray ionization‐mass spectrometry method. For elucidating intestinal transport mechanism, the cells were incubated with or without transport inhibitors, energy source, or a metabolic inhibitor. Phloridzin and verapamil as inhibitors of sodium glucose transport protein (SGLT1) and P‐glycoprotein, respectively, were not responsible for cellular uptake of SMM. Glucose and sodium azide were not affected by the cellular accumulation of SMM. The efflux ratio of SMM was 0.26, implying that it is not effluxed through Caco‐2 cells. The apparent coefficient permeability (P _app) of SMM was 4.69 × 10^?5 cm/s, indicating that it will show good oral absorption in in vivo .