Oat-enriched diet reduces inflammatory status assessed by circulating cell-derived microparticle concentrations in type 2 diabetes

This work was funded by the Chief Scientists Office of the Scottish Government by a joint grant to the University of the Highland and Islands, Grampian Health Board, Biomathematics and Statistics Scotland and the Rowett Institute of Nutrition and Health, University of Aberdeen. Additional support was provided by Provexis plc.Peer reviewedPublisher PD

Translocation of Crohn's disease Escherichia coli across M-cells: contrasting effects of soluble plant fibres and emulsifiers

Background Crohns disease is common in developed nations where the typical diet is low in fibre and high in processed food. Primary lesions overlie Peyers patches and colonic lymphoid follicles where bacterial invasion through M-cells occurs. We have assessed the effect of soluble non-starch polysaccharide (NSP) and food emulsifiers on translocation of Escherichia coli across M-cells. Methods To assess effects of soluble plant fibres and food emulsifiers on translocation of mucosa-associated E coli isolates from Crohns disease patients and from non-Crohns controls, we used M-cell monolayers, generated by co-culture of Caco2-cl1 and Raji B cells, and human Peyers patches mounted in Ussing chambers. Results E coli translocation increased across M-cells compared to parent Caco2-cl1 monocultures; 15.8-fold (IQR 6.2-32.0) for Crohns disease E coli (N=8) and 6.7-fold (IQR 3.7-21.0) for control isolates (N=5). Electronmicroscopy confirmed E coli within M-cells. Plantain and broccoli NSP markedly reduced E coli translocation across M-cells at 5 mg/ml (range 45.3-82.6% inhibition, pandlt;0.01); apple and leek NSP had no significant effect. Polysorbate-80, 0.01% vol/vol, increased E coli translocation through Caco2-cl1 monolayers 59-fold (pandlt;0.05) and, at higher concentrations, increased translocation across M-cells. Similarly, E coli translocation across human Peyers patches was reduced 45+/-7% by soluble plantain NSP (5 mg/ml) and increased 2-fold by polysorbate-80 (0.1% vol/vol). Conclusions Translocation of E coli across M-cells is reduced by soluble plant fibres, particularly plantain and broccoli, but increased by the emulsifier Polysorbate-80. These effects occur at relevant concentrations and may contribute to the impact of dietary factors on Crohns disease pathogenesis

Dietary Supplementation with Soluble Plantain Non-Starch Polysaccharides Inhibits Intestinal Invasion of Salmonella Typhimurium in the Chicken

Soluble fibres (non-starch polysaccharides, NSP) from edible plants but particularly plantain banana (Musa spp.), have been shown in vitro and ex vivo to prevent various enteric pathogens from adhering to, or translocating across, the human intestinal epithelium, a property that we have termed contrabiotic. Here we report that dietary plantain fibre prevents invasion of the chicken intestinal mucosa by Salmonella. In vivo experiments were performed with chicks fed from hatch on a pellet diet containing soluble plantain NSP (0 to 200 mg/d) and orally infected with S.Typhimurium 4/74 at 8 d of age. Birds were sacrificed 3, 6 and 10 d post-infection. Bacteria were enumerated from liver, spleen and caecal contents. In vitro studies were performed using chicken caecal crypts and porcine intestinal epithelial cells infected with Salmonella enterica serovars following pre-treatment separately with soluble plantain NSP and acidic or neutral polysaccharide fractions of plantain NSP, each compared with saline vehicle. Bacterial adherence and invasion were assessed by gentamicin protection assay. In vivo dietary supplementation with plantain NSP 50 mg/d reduced invasion by S.Typhimurium, as reflected by viable bacterial counts from splenic tissue, by 98.9% (95% CI, 98.1–99.7; P<0.0001). In vitro studies confirmed that plantain NSP (5–10 mg/ml) inhibited adhesion of S.Typhimurium 4/74 to a porcine epithelial cell-line (73% mean inhibition (95% CI, 64–81); P<0.001) and to primary chick caecal crypts (82% mean inhibition (95% CI, 75–90); P<0.001). Adherence inhibition was shown to be mediated via an effect on the epithelial cells and Ussing chamber experiments with ex-vivo human ileal mucosa showed that this effect was associated with increased short circuit current but no change in electrical resistance. The inhibitory activity of plantain NSP lay mainly within the acidic/pectic (homogalacturonan-rich) component. Supplementation of chick feed with plantain NSP was well tolerated and shows promise as a simple approach for reducing invasive salmonellosis

Cell wall associated phenylpropanoids in fruit and vegetables

Low molecular weight phenylpropanoid components of plant cell walls, ingested by humans as dietary fruit and vegetables, are released from covalent linkages to the cell wall by bacterial fermentation in the lower gut.  Based on their structural characteristics, such phenylpropanoids and their microbial metabolites potentially have the capacity to interact specifically with the cyclooxgenase (COX) enzymes, important mediators of the inflammation associated with developing neoplasia.  These cell wall components could therefore constitute an important dietary source of potentially bioactive compounds. A combination of enzymatic and chemical cell wall hydrolysis was used to liberate covalently bound low molecular weight phenylpropanoid derivatives from the polymer matrix of fruit and vegetable cell walls.  A range of ester- and ether-linked hydroxycinnamic and benzoic acids and derivatives, including significant quantities of dimeric 4-hydroxy-3-methoxycinnamates, was recovered from all samples examined.  An estimate of the bioavailability of these cell wall components was made by quantifying the proportion of measured cell wall phenylpropanoids released from the wall by enzyme digestion.  In approximately one half of the plant foods examined, these cell wall components were shown to occur at levels which represent a significant dietary burden (based on measured bioavailable compounds in a serving of 125 g plant fresh weight), and thus their presence in the diet is generally underestimated. The major cell wall associated hydroxycinnamates and the 4-hydroxy-3-methoxycinnamate dehydrodimers, as well as their major microbial metabolic products the phenylpropanoates, were examined for COX - inhibitory activity using an enzyme immunoassay.  Results showed a broad spectrum of activities, mainly non-specific.  Cell wall phenylpropanoid derivatives thus have the potential to alter the pattern of prostanoid formation in individuals consuming, as part of their diet, food plants with a high cell wall hydroxycinnamate content.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Platelet hyperactivity in COVID-19: Can the tomato extract fruitflow® be used as an antiplatelet regime?

The ongoing coronavirus disease 2019 (COVID-19) pandemic caused by the SARS-CoV-2 virus is now considered a global public health threat. The primary focus has been on reducing the viral spread and treating respiratory symptoms; as time goes on, the impact of COVID-19 on neurological and haemostatic systems becomes more evident. The clinical data suggest that platelet hyperactivity plays a role in the pathology of COVID-19 from its onset and that platelets may serve critical functions during COVID‐19 progression. Hyperactivation of blood platelets and the coagulation system are emerging as important drivers of inflammation and may be linked to the severity of the ‘cytokine storm’ induced in severe cases of COVID-19, in which disseminated intravascular coagulation, and platelet hyperactivity are associated with poor prognosis and increased risk of mortality. We propose that targeting platelet hyperactivity in the early stages of COVID-19 infection may reduce the immunothrombotic complications of COVID-19 and subdue the systemic inflammatory response. Lowering baseline platelet activity may be of particular importance for higher-risk groups. As an alternative to antiplatelet drugs, an inappropriate intervention in public health, we propose that the dietary antiplatelet agent Fruitflow®, derived from tomatoes, may be considered a suitable therapy. Fruitflow® contains antiplatelet and anti-inflammatory compounds that target the mechanisms of platelet activation specific to COVID-19 and can be considered a safe and natural antiplatelet regime

Differential internalin A levels in biofilms of Listeria monocytogenes grown on different surfaces and nutrient conditions

Listeria monoctyogenes is a foodborne pathogen containing the surface protein, internalin A (InlA). The expression of this protein permits the invasion of L. monocytogenes into intestinal epithelial cells expressing the receptor E-cadherin, thus crossing the intestinal barrier and resulting in listerosis. The main aim of this work was to investigate InlA levels in different L. monocytogenes strains in both planktonic and sessile states using an anti-InlA antibody. Biofilms were grown in high and low nutrient environments on glass, stainless steel and polytetrafluoroethylene (PTFE). This study demonstrated that InlA levels varied greatly between strains and serotypes of L. monocytogenes. However, the serotypes 1/2a, 1/2b and 4b, associated with the largest number of outbreaks of listerosis consistently showed the highest InlA levels, regardless of nutrient content or planktonic or sessile state. Differences in InlA levels were also observed in biofilms grown on different surfaces such as glass, stainless steel and PTFE, with a significant reduction in InlA levels observed in biofilms on PTFE. Interestingly, although a large number of the total cells observed in biofilms formed in tap-water were non-cultivable, the virulence factor, InlA, was expressed at levels between 78 and 85%, thus indicating that these cells may still be virulent. A greater understanding of the factors that affect the levels of InlA on the surface of L. monocytogenes, is essential in the appreciation of the role of InlA in the persistence of biofilms containing L. monocytogenes and their potential to cause food borne disease

Anti-platelet effects of olive oil extract: In vitro functional and proteomic studies

Purpose: Platelets play a key role in haemostasis and wound healing, contributing to formation of vascular plugs. They are also involved in formation of atherosclerosic plaques. Some traditional diets, like the Mediterranean diet, are associated with a lower risk of cardiovascular disease. Components in these diets may have anti-platelet functions contributing to their health benefits. Methods: We studied the effects of alperujo extract, an olive oil production waste product containing the majority of polyphenols found in olive fruits, through measurement of effects on platelet aggregation and activation in isolated human platelets, and through identification of changes in the platelet proteome. Results: Alperujo extract (40 mg/L) significantly decreased in vitro ADP- (p = 0.002) and TRAP- (p = 0.02) induced platelet activation as measured by the flow cytometry using the antibody for p-selectin (CD62p), but it did not affect the conformation of the fibrinogen receptor as measured by flow cytometry using the antibodies for anti-fibrinogen, CD42a and CD42b. Alperujo extract (100 mg/L) inhibited both collagen- and TRAP-induced platelet aggregation by 5% (p < 0.05), and a combination of hydroxytyrosol and 3,4-dihydroxyphenylglycol were, at least partly, responsible for this effect. Proteomic analysis identified nine proteins that were differentially regulated by the alperujo extract upon ADP-induced platelet aggregation. These proteins represent important mechanisms that may underlie the anti-platelet effects of this extract: regulation of platelet structure and aggregation, coagulation and apoptosis, and signalling by integrin αIIb/β3. Conclusions: Alperujo extract may protect against platelet activation, platelet adhesion and possibly have anti-inflammatory properties. © 2010 Springer-Verlag.Peer Reviewe

Soluble plantain NSP increases the transmucosal short circuit current of <i>ex vivo</i> human ileal follicle-associated epithelium.

<p>Plantain NSP (5 mg/mL) significantly increased (<b>A</b>) transmucosal short circuit current (<i>I_sc</i>), with a concomitant decrease in (<b>B</b>) epithelial potential difference (PD, apical-side negative), during pre-treatment of, and blockade of translocation of S. Typhimurium across <i>ex vivo</i> human ileal follicle-associated epithelium (FAE) mounted in Ussing chambers. (<b>C</b>) Trans-epithelial electrical resistance (TEER) was maintained throughout the experiment. N = 4, with 2 tissue replicates in each case. * <i>P</i><0.05, ** <i>P</i><0.01, ANOVA. For each tissue, <i>I</i>_sc and PD responses were calculated and expressed as the increment change (Δ) for each sampling period. Arrows at T₀ min indicate addition of EGFP-expressing <i>S.</i> Typhimurium LT2 to the mucosal compartment (1×10⁸ CFU/mL). Overnight culture of Ussing chamber serosal medium following 2 h infection had already demonstrated soluble plantain fibre to block translocation of <i>Salmonella</i> across isolated human FAE in this experiment; see reference <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087658#pone.0087658-Roberts2" target="_blank">[21]</a>.</p

Dietary supplementation with soluble plantain NSP reduces chick salmonellosis <i>in vivo.</i>

<p>(<b>A</b>) Following infection of 8 day-old inbred specified pathogen-free White Leghorn Line 0 chicks with <i>S.</i> Typhimurium 4/74, soluble plantain NSP supplementation of a commercial pellet feed significantly reduced bacterial numbers found in the spleen 3 d post-infection. (<b>B</b>) Supplementation with soluble plantain NSP had little significant effect on presence of <i>S.</i> Typhimurium CFU cultured from the liver, excepting at 10 d post infection. (<b>C</b>) Total CFU observed within the caecal lumen were relatively unchanged at all doses of plantain NSP supplementation compared to birds fed a control diet. Significant differences from control (non-supplemented NSP) diet, * <i>P</i><0.05; ** <i>P</i><0.01; *** <i>P</i><0.0001 Kruskal-Wallis (N = 4–7 birds, n = 2 replicates).</p

Soluble plantain NSP inhibits adhesion of <i>S</i>. Typhimurium to the porcine B1OXI enterocyte cell-line <i>in vitro</i>.

<p>Pre-treatment (30 min) with soluble plantain NSP, dose-dependently blocked adhesion of (<b>A</b>) <i>S</i>. Typhimurium LT2 and (<b>B</b>) <i>S</i>. Typhimurium 4/74 to B1OXI cells (N = 3 experiments, each with n = 4 replicates); * <i>P</i><0.05, ** <i>P</i><0.01, *** <i>P</i><0.001, Kruskal-Wallis. Invasion of (<b>C</b>) <i>S.</i> Typhimurium LT2 and (<b>D</b>) <i>S.</i> Typhimurium 4/74 into B1OXI cells was also blocked by plantain NSP. Data (mean ± SEM) expressed relative to adherence (or invasion) of vehicle-treated control (100%). Light microscopy of Giemsa-stained B1OXI cells and <i>S</i>. Typhimurium 4/74 in absence (<b>E</b>) or presence (<b>F</b>) of 10 mg/mL soluble plantain NSP. Arrows indicate bacteria.</p