Ex Vivo Colonic Fermentation of NUTRIOSE® Exerts Immuno-Modulatory Properties and Strong Anti-Inflammatory Effects.

NUTRIOSE® (Roquette, Lestrem, France) is a resistant dextrin with well-established prebiotic effects. This study evaluated the indirect effects of pre-digested NUTRIOSE® on host immune response and gut barrier integrity. Fecal samples from eight healthy donors were inoculated in a Colon-on-a-plate® system (ProDigest, Ghent, Belgium) with or without NUTRIOSE® supplementation. Following 48 h fermentation, colonic suspensions were tested in a Caco-2/THP1-Blue™ co-culture system to determine their effects on gut barrier activity (transepithelial electrical resistance) and immune response following lipopolysaccharide stimulation. Additionally, changes in short-chain fatty acid levels (SCFA) and microbial community composition following a 48 h fermentation in the Colon-on-a-plate® system were measured. Across all donors, immune-mediated intestinal barrier damage was significantly reduced with NUTRIOSE®-supplemented colonic suspensions versus blank. Additionally, IL-6 and IL-10 levels were significantly increased, and the level of the neutrophil chemoattractant IL-8 was significantly decreased with NUTRIOSE®-supplemented colonic suspensions versus blank in the co-culture models following lipopolysaccharide stimulation. These beneficial effects of NUTRIOSE® supplementation were likely due to increased acetate and propionate levels and the enrichment of SCFA-producing bacteria. NUTRIOSE® was well fermented by the colonic bacteria of all eight donors and had protective effects on inflammation-induced disruption of the intestinal epithelial barrier and strong anti-inflammatory effects.

Bridging preclinical and clinical gut microbiota research using the ex vivo SIFR® technology.

Introduction: While modulation of the human adult gut microbiota is a trending strategy to improve health, the underlying mechanisms are poorly understood. Methods: This study aimed to assess the predictive value of the ex vivo, reactor-based, high-throughput SIFR® (Systemic Intestinal Fermentation Research) technology for clinical findings using three structurally different prebiotics [inulin (IN), resistant dextrin (RD) and 2'-fucosyllactose (2'FL)]. Results: The key finding was that data obtained within 1-2 days were predictive for clinical findings upon repeated prebiotic intake over weeks: among hundreds of microbes, IN stimulated Bifidobacteriaceae, RD boosted Parabacteroides distasonis, while 2'FL specifically increased Bifidobacterium adolescentis and Anaerobutyricum hallii. In line with metabolic capabilities of these taxa, specific SCFA (short-chain fatty acids) were produced thus providing insights that cannot be obtained in vivo where such metabolites are rapidly absorbed. Further, in contrast to using single or pooled fecal microbiota (approaches used to circumvent low throughput of conventional models), working with 6 individual fecal microbiota enabled correlations that support mechanistic insights. Moreover, quantitative sequencing removed the noise caused by markedly increased cell densities upon prebiotic treatment, thus allowing to even rectify conclusions of previous clinical trials related to the tentative selectivity by which prebiotics modulate the gut microbiota. Counterintuitively, not the high but rather the low selectivity of IN caused only a limited number of taxa to be significantly affected. Finally, while a mucosal microbiota (enriched with Lachnospiraceae) can be integrated, other technical aspects of the SIFR® technology are a high technical reproducibility, and most importantly, a sustained similarity between the ex vivo and original in vivo microbiota. Discussion: By accurately predicting in vivo results within days, the SIFR® technology can help bridge the so-called "Valley of Death" between preclinical and clinical research. Facilitating development of test products with better understanding of their mode of action could dramatically increase success rate of microbiome modulating clinical trials.Graphical Abstract.

Diet Supplementation with NUTRIOSE, a Resistant Dextrin, Increases the Abundance of Parabacteroides distasonis in the Human Gut.

SCOPE: An imbalance of the gut microbiota ("dysbiosis") is associated with numerous chronic diseases, and its modulation is a promising novel therapeutic approach. Dietary supplementation with soluble fiber is one of several proposed modulation strategies. This study aims at confirming the impact of the resistant dextrin NUTRIOSE (RD), a soluble fiber with demonstrated beneficial health effects, on the gut microbiota of healthy individuals. METHODS AND RESULTS: Fifty healthy women are enrolled and supplemented daily with either RD (n = 24) or a control product (n = 26) during 6 weeks. Characterization of the fecal metagenome with shotgun sequencing reveals that RD intake dramatically increases the abundance of the commensal bacterium Parabacteroides distasonis. Furthermore, presence in metagenomes of accessory genes from P. distasonis, coding for susCD (a starch-binding membrane protein complex) is associated with a greater increase of the species. This suggests that response to RD might be strain-dependent. CONCLUSION: Supplementation with RD can be used to specifically increase P. distasonis in gut microbiota of healthy women. The magnitude of the response may be associated with fiber-metabolizing capabilities of strains carried by subjects. Further research will seek to confirm that P. distasonis directly modulates the clinical effects observed in other studies.

The ALGOVUE Clinical Trial: Effects of the Daily Consumption of Eggs Enriched with Lutein and Docosahexaenoic Acid on Plasma Composition and Macular Pigment Optical Density.

BACKGROUND: Carotenoids and docosahexaenoic acid (DHA) were identified as essential components for eye health and are both naturally present in eggs. OBJECTIVE: We aimed to evaluate the effect of the daily consumption of two eggs enriched with lutein/zeaxanthin and DHA on macular pigment optical density (MPOD) and on circulating xanthophyll and fatty acid concentrations in healthy participants. METHODS: Ninety-nine healthy volunteers consumed either two standard eggs or two enriched eggs per day for 4 months. MPOD was measured at baseline (V0) and at follow-up (V4) using a modified confocal scanning laser ophthalmoscope (primary outcome). Blood samples were collected to determine total plasma and lipoprotein fatty acids and lutein/zeaxanthin compositions at V0 and V4 (secondary outcomes). RESULTS: A slight but significant increase in MPOD was observed for all study participants consuming two eggs per day for 4 months at all eccentricities (0.5°, 1°, 2°, and 4°). Plasma and lipoprotein lutein, zeaxanthin, and DHA concentrations significantly increased in both groups but were greater in the enriched group (for the enriched group (V0 vs. V4): lutein, 167 vs. 369 ng/mL; zeaxanthin, 17.7 vs. 29.2 ng/mL; DHA, 1.89 vs. 2.56% of total fatty acids). Interestingly, lutein from high-density lipoprotein (HDL) was strongly correlated with MPOD at 0.5 and 1° eccentricities (rho = 0.385, p = 0.008, and rho = 0.461, p = 0.001, respectively). CONCLUSIONS: MPOD was slightly increased in both groups. Lutein, zeaxanthin, and DHA plasma concentrations were strongly enhanced in the enriched group compared with the standard group. A significant correlation was found between MPOD level and lutein concentration in HDL.

Impact of dietary supplementation with resistant dextrin (NUTRIOSE®) on satiety, glycaemia, and related endpoints, in healthy adults.

PURPOSE: Resistant dextrin (RD) supplementation has been shown to alter satiety, glycaemia, and body weight, in overweight Chinese men; however, there are limited data on its effects in other demographic groups. Here, we investigated the effects of RD on satiety in healthy adults living in the United Kingdom. METHODS: 20 normal weight and 16 overweight adults completed this randomised controlled cross-over study. Either RD (14 g/day NUTRIOSE® FB06) or maltodextrin control was consumed in mid-morning and mid-afternoon preload beverages over a 28-day treatment period with crossover after a 28-day washout. During 10-h study visits (on days 1, 14, and 28 of each treatment period), satietogenic, glycaemic and anorectic hormonal responses to provided meals were assessed. RESULTS: Chronic supplementation with RD was associated with higher fasted satiety scores at day 14 (P = 0.006) and day 28 (P = 0.040), compared to control. RD also increased satiety after the mid-morning intervention drink, but it was associated with a reduction in post-meal satiety following both the lunch and evening meals (P < 0.01). The glycaemic response to the mid-morning intervention drink (0-30 min) was attenuated following RD supplementation (P < 0.01). Whilst not a primary endpoint we also observed lower systolic blood pressure at day 14 (P = 0.035) and 28 (P = 0.030), compared to day 1, following RD supplementation in the normal weight group. Energy intake and anthropometrics were unaffected. CONCLUSIONS: RD supplementation modified satiety and glycaemic responses in this cohort, further studies are required to determine longer-term effects on body weight control and metabolic markers. CLINICALTRIALS. GOV REGISTRATION: NCT02041975 (22/01/2014).

The Impact of Maltitol-Sweetened Chewing Gum on the Dental Plaque Biofilm Microbiota Composition.

Background: The oral cavity harbors a complex microbial ecosystem, intimately related to oral health and disease. The use of polyol-sweetened gum is believed to benefit oral health through stimulation of salivary flow and impacting oral pathogenic bacteria. Maltitol is often used as sweetener in food products. This study aimed to establish the in vivo effects of frequent consumption of maltitol-sweetened chewing gum on the dental plaque microbiota in healthy volunteers and to establish the cellular and molecular effects by in vitro cultivation and transcriptional analysis. Results: An intervention study was performed in 153 volunteers, randomly assigned to three groups (www.trialregister.nl; NTR4165). One group was requested to use maltitol gum five times daily, one group used gum-base, and the third group did not use chewing gum. At day 0 and day 28, 24 h-accumulated supragingival plaque was collected at the lingual sites of the lower jaw and the buccal sites of the upper jaw and analyzed by 16S ribosomal rRNA gene sequencing. At day 42, 2 weeks after completion of the study, lower-jaw samples were collected and analyzed. The upper buccal plaque microbiota composition had lower bacterial levels and higher relative abundances of (facultative) aerobic species compared to the lower lingual sites. There was no difference in bacterial community structure between any of the three study groups (PERMANOVA). Significant lower abundance of several bacterial phylotypes was found in maltitol gum group compared to the gum-base group, including Actinomyces massiliensis HOT 852 and Lautropia mirabilis HOT 022. Cultivation studies confirmed growth inhibition of A. massiliensis and A. johnsonii by maltitol at levels of 1% and higher. Transcriptome analysis of A. massiliensis revealed that exposure to maltitol resulted in changes in the expression of genes linked to osmoregulation, biofilm formation, and central carbon metabolism. Conclusion: The results showed that chewing itself only marginally impacted the plaque microbiota composition. Use of maltitol-sweetened gum lowered abundance of several bacterial species. Importantly, the species impacted play a key role in the early formation of dental biofilms. Further studies are required to establish if frequent use of maltitol gum impacts early dental-plaque biofilm development.

Effect of maltitol-containing chewing gum use on the composition of dental plaque microbiota in subjects with active dental caries.

Background: Sugar alcohols such as xylitol are incorporated in a number of oral hygiene products for their anti-cariogenic properties while chewing gum is known to be beneficial to oral hygiene. Objective: The aim of this study was to determine the composition of the dental plaque microbiota in patients with active caries before and after using a chewing gum supplemented with maltitol. Design : Forty subjects with active caries were randomly allocated to chew maltitol gum or gum base for two weeks. A healthy control group used gum base for two weeks. Plaque samples were collected before and after treatment and the microbiota analysed by pyrosequencing of 16S rRNA genes. Results : A total of 773,547 sequences were obtained from 117 samples. There was no difference in structure of the bacterial communities between groups (AMOVA). There was a significant difference in community membership between groups, (AMOVA, p=0.009). There was a significant difference between the control group after treatment and the maltitol patient group after treatment (p<0.001). A. naeslundii HOT-176 and Actinomyces HOT-169 were significantly reduced following use of maltitol chewing gum in patients. Conclusions : This study has shown that chewing gum containing maltitol had minor effects on the composition of the plaque microbiome.

Incorporation of lutein and docosahexaenoic acid from dietary microalgae into the retina in quail.

Lutein and docosahexaenoic acid (DHA) are associated with the prevention of age-related macular degeneration (AMD). Since microalgae are potent natural sources of these nutrients, their nutritional value should be evaluated based on the bioavailability of lutein and DHA for the retina via the plasmatic compartment. In this study, quail were fed for 5 months either with a diet supplemented or deprived with microalgae rich in lutein and DHA. In the microalgae-fed group, the retinal concentrations of lutein and zeaxanthin gradually increased whereas in plasma, these compounds started to increase from the first month of supplementation. We also observed a significant increase in retinal and plasmatic levels of DHA in the microalgae-fed group. In conclusion, the plasmatic and retinal contents of lutein and DHA were significantly increased in quail fed with lutein- and DHA-rich microalgae. Food fortification with microalgae may be an innovative way to increase lutein and DHA consumption in humans.

Lipid transport function is the main target of oral oleoylethanolamide to reduce adiposity in high-fat-fed mice.

We evaluated the biological basis of reduced fat gain by oleoylethanolamide (OEA) in high-fat-fed mice and sought to determine how degradation of OEA affected its efficiency by comparing its effects to those of KDS-5104, a nonhydrolyzable lipid OEA analog. Mice were given OEA or KDS-5104 by the oral route (100 mg/kg body weight). Sixty-eight variables per mouse, describing six biological processes (lipid transport, lipogenesis, energy intake, energy expenditure, endocannabinoid signaling, and glucose metabolism), spanning gene expression of biochemical and physiological parameters were examined to determine the primary target whereby OEA reduces fat gain. Although KDS-5104 but not OEA was resistant to fatty acid amide hydrolase hydrolysis, OEA was degraded by an unidentified hydrolysis system in the liver. Nevertheless, both compounds equally decreased body fat pads after 5 weeks (20%; P < 0.05). The six biological functions constructed from the 68 initial variables predicted up to 58% of adipose fat variations. Lipid transport appeared central to the explanation for body fat deposition (16%; P < 0.0001), in which decreased expression of the FAT/CD36 gene was the component most related to adipose depots. Lipid transport appears to be a determinant player in the OEA fat-lowering response, with adipose tissue FAT/CD36 expression being the most relevant bioindicator of OEA action.

No observable differences in glycemic response to maltitol in human subjects from 3 ethnically diverse groups.

Consumption of a low glycemic index (GI) diet may be helpful in the management and prevention of diabetes and cardiovascular disease. The investigation of GI has been predominantly confined to white subjects. We hypothesized that differences in glycemic response (GR) may be observable in subjects of different ethnic origin. The objective of the present study was to determine GR to a high GI (glucose) and low GI (maltitol) test drink in subjects of different ethnic origin. In a randomized, single-blind crossover trial, 10 whites, 10 South Indians and 10 Chinese subjects consumed either glucose or maltitol test drink containing 50 g of one of the test products on different occasions. Capillary blood glucose samples were taken at 15 and 10 minutes before and 0, 15, 30, 45, 60, 90, 120, 150, and 180 minutes after consumption of the test drink. The incremental area under the curve of glucose and maltitol were not significantly different between the 3 groups. The mean GR for maltitol was 33.5% in whites, 32.9% in Chinese, and 23.1% in South Indians. The results presented here confirmed that there are no observable differences noted in GR to a high-GI or low-GI test drink between the 3 ethnically diverse groups. We conclude that different ethnic groups exhibit similar GR to low- and high-GI drinks, and GR to maltitol is similar irrespective of ethnic background.

Short-term digestive tolerance of chocolate formulated with maltitol in children.

INTRODUCTION: Polyols are molecules of interest for food industries because of their technological and nutritional properties. Maltitol is known for its non-acidogenic and low-energetic properties. Our primary objective was to evaluate the digestive tolerance of maltitol in children. The secondary objective was to compare the organoleptic properties of maltitol and sucrose in chocolate. METHOD: Healthy children were included in a double-blind, randomized parallel study versus placebo. The subjects received one dose of either maltitol or sucrose chocolate per week. Increasing doses were tested from 5 to 15 g maltitol in chocolate. Abdominal pain, rumbling, bloating and flatulence scores were evaluated using visual analog scales. RESULTS: Some statistical differences on intestinal parameters were observed in the maltitol group compared with placebo, mainly concerning flatulence scores. Nevertheless, these scores remained low and could be considered minor. CONCLUSION: Our results suggest that maltitol was well tolerated in children at 15 g in one intake.

Analysis of gene expression pattern reveals potential targets of dietary oleoylethanolamide in reducing body fat gain in C3H mice.

Oleoylethanolamide (OEA) has been previously reported to regulate food intake and body weight gain when administered intraperitoneally. Nevertheless, little information is available with regard to oral administration. To assess whether oral OEA can also exert a similar effect on body fat, we fed C3H mice a high-fat diet supplemented with either 10 or 100 mg/kg body weight OEA for 4 weeks. OEA supplementation significantly lowered food intake over the 4 weeks and decreased adipose tissue mass. Plasma triglyceride levels were also significantly decreased by OEA treatment. In order to identify the potential molecular targets of OEA action, we screened the expression levels of 44 genes related to body fat mass and food intake in peripheral tissues. Adipose tissue fatty acid amide hydrolase (FAAH), intestinal fatty acid transporter/cluster of differentiation 36 and the OEA receptor G-protein-coupled receptor 119 (GPR119) were among the most OEA-responsive genes. They were also associated with reduced body fat pads regardless of the dose. Adipose FAAH was found to be primarily associated with a decrease in food intake. Our data suggest that the anti-obesity activity of OEA partially relies on modulation of the FAAH pathway in adipose tissue. Another mechanism might involve modulation of the newly discovered GPR119 OEA signaling pathway in the proximal intestine. In conclusion, our study indicates that oral administration of OEA can effectively decrease obesity in the mouse model and that modulation of the endocannabinoid fatty acid ethanolamide pathway seems to play an important role both in adipose tissue and in small intestine.

Biological functions and metabolism of oleoylethanolamide.

The present review is focused on the metabolism and the emerging roles of oleoylethanolamide (OEA) with emphasis on its effects on food intake control and lipid metabolism. The biological mechanism of action, including a non-genomic effect mediated through peroxisome proliferator-activated receptor alpha (PPAR-alpha) and transient receptor potential vanilloid type 1 (TRPV1) receptor, is discussed. The research related to fatty acid ethanolamides has been focused until recently on anandamide and its interaction with cannabinoid receptor subtype 1. The roles of other N-acyl ethanolamine fatty acid derivatives have been neglected until it was demonstrated that OEA can modulate food intake control through interaction with PPAR-alpha. Further investigations demonstrated that OEA modulates lipid and glucose metabolism, and recent study confirmed that OEA is an antagonist of TRVP1. It has been demonstrated that OEA has beneficial effects on health by inducing food intake control, lipid beta-oxidation, body weight loss and analgesic effects. The investigation of the mechanism of action revealed that OEA activates PPAR-alpha and stimulates the vagal nerve through the capsaicin receptor TRPV1. Pre-clinical studies showed that OEA remains active when administered orally.

Analysis of chemically synthesized oleoylethanolamide by gas-liquid chromatography.

Oleoylethanolamide (OEA) is known to potentially have beneficial biological effects on weight management by controlling food intake and activating lipid catabolism. In biological fluids, OEA and other endogenously biosynthesized fatty acid ethanolamides are usually analyzed by liquid chromatography-mass spectrometry (LC-MS). The present study provides analytical method to routinely assess the quality of OEA prepared for biological studies by gas-liquid chromatography (GLC). The preparation of OEA for biomedical studies can be performed by N-acylation of oleic acid/esters or using oleoyl chloride. In the present study, OEA was prepared by transamidation of triolein. The analysis of the synthesized OEA has been performed by gas-liquid chromatography of its trimethylsilyl ether (TMS) derivatives. Free OEA cannot be analyzed as such because dehydration of the ethanolamide moiety promptly happens in the GLC injection. This thermal degradation reaction gives rise to the formation of an oxazoline derivative. The TMS moiety prevents the reaction, and the structure of the formed derivative was assessed by mass spectrometry. We show here that OEA prepared for biological studies can be routinely analyzed by GLC after TMS derivative preparation.