Butyrate enemas upregulate Muc genes expression but decrease adherent mucus thickness in mice colon.

Colonic mucosal protection is provided by the mucus gel, mainly composed of mucins. Several factors can modulate the formation and the secretion of mucins, and among them butyrate, an end-product of carbohydrate fermentation. However, the specific effect of butyrate on the various colonic mucins, and the consequences in terms of the mucus layer thickness are not known. Our aim was to determine whether butyrate modulates colonic MUC genes expression in vivo and whether this results in changes in mucus synthesis and mucus layer thickness. Mice received daily for 7 days rectal enemas of butyrate (100 mM) versus saline. We demonstrated that butyrate stimulated the gene expression of both secreted (Muc2) and membrane-linked (Muc1, Muc3, Muc4) mucins. Butyrate especially induced a 6-fold increase in Muc2 gene expression in proximal colon. However, butyrate enemas did not modify the number of epithelial cells containing the protein Muc2, and caused a 2-fold decrease in the thickness of adherent mucus layer. Further studies should help understanding whether this last phenomenon, i.e. the decrease in adherent mucus gel thickness, results in a diminished protective function or not.

MUC genes are differently expressed during onset and maintenance of inflammation in dextran sodium sulfate-treated mice.

Colonic mucosal protection is provided by mucous gel, mainly composed of secreted (Muc2) and membrane-bound (Muc1, Muc3, Muc4) mucins. Our aim was to determine the expression profile of secreted and membrane-bound mucins in experimental dextran sulfate sodium (DSS)-induced colitis. Acute colitis was induced in Balb/C mice by oral administration of 1.0% DSS (5 days) and chronic colitis was maintained by subsequent 0.15% DSS treatment (28 days). Clinical symptoms (mortality, weight gain), stool scores, and MPO activity confirmed the inflammatory state in the two phases of colitis. Muc2 gene expression was not modified by colitis, whereas Muc3 gene expression was increased (x2) only in the cecum and the distal colon of mice after acute colitis. Muc1 and Muc4 mRNA levels were more significantly increased in the cecum (x8-10) than in colonic segments (x4) after acute colitis. TFF3 involved in mucosal repair was up-regulated during colitis induction. These results indicate that Muc and TFF3 genes are regulated early in inflammation and suggest that their mRNA levels could be used as early markers of inflammation.

Butyrate specifically modulates MUC gene expression in intestinal epithelial goblet cells deprived of glucose.

The mucus layer covering the gastrointestinal mucosa is considered the first line of defense against aggressions arising from the luminal content. It is mainly composed of high molecular weight glycoproteins called mucins. Butyrate, a short-chain fatty acid produced during carbohydrate fermentation, has been shown to increase mucin secretion. The aim of this study was to test 1) whether butyrate regulates the expression of various MUC genes, which are coding for protein backbones of mucins, and 2) whether this effect depends on butyrate status as the major energy source of colonocytes. Butyrate was provided at the apical side of human polarized colonic goblet cell line HT29-Cl.16E in glucose-rich or glucose-deprived medium. In glucose-rich medium, butyrate significantly increased MUC3 and MUC5B expression (1.6-fold basal level for both genes), tended to decrease MUC5AC expression, and had no effect on MUC2 expression. In glucose-deprived medium, i.e., when butyrate was the only energy source available, MUC3 and MUC5B increase persisted, whereas MUC5AC expression was significantly enhanced (3.7-fold basal level) and MUC2 expression was strikingly increased (23-fold basal level). Together, our findings show that butyrate is able to upregulate colonic mucins at the transcriptional level and even better when it is the major energy source of the cells. Thus the metabolism of butyrate in colonocytes is closely linked to some of its gene-regulating effects. The distinct effects of butyrate according to the different MUC genes could influence the composition and properties of the mucus gel and thus its protective function.

Development of an in vitro system simulating bucco-gastric digestion to assess the physical and chemical changes of food.

The release of nutrients from solid food depends on the physical and chemical characteristics of substrates, and on dynamic physiological events including pH, gastric emptying and enzymatic secretion. Our laboratory has developed an in vitro digestive system mimicking mouth and stomach processes to determine physical and chemical changes of bread during digestion. To simulate oral-phase digestion, bread was minced and subjected to in vitro amylase digestion, releasing 219 +/- 11 g oligosaccharides/kg total carbohydrate. During the gastric phase, bread proteins, which are converted into insoluble aggregated proteins during breadmaking, were emptied in various states of peptic digestion: undigested aggregated proteins and degraded proteins of intermediate and low molecular weight. The mean particle size of ground bread decreased progressively to the end of the gastric digestion (from 292 to 109 microm). The in vitro digestive system proved to be a useful tool for understanding the dynamic digestion of various food components held within the structure of a food matrix.

Particle size of solid food after human mastication and in vitro simulation of oral breakdown.

Mastication, the first step in food digestion, results in the breakdown of solid food and its lubrication with saliva. Although the rate and extent of starch digestion are closely dependent on the way food is chewed, this factor has not been adequately considered in the preparation of food for in vitro digestion experiments. The purpose of this study was to determine the size distribution of starchy food particles before swallowing and to use an in vitro mincing procedure to simulate how food is divided up during chewing. Foods differing in texture and size (bread, spaghetti and tortiglioni) were chewed by 12 healthy subjects and spat out before swallowing. Chewing time and saliva impregnation were measured for each mouthful. The particle sizes resulting from experiments with chewed and minced bread and pasta were analysed respectively by light laser diffraction and image analysis. Chewing time was longer for bread than pasta, resulting in higher saliva impregnation. Chewed bread showed a bimodal distribution of particle size (30 microns, 500 microns), whereas both kinds of pasta produced particles of similar size (0.5 to 30 mm2) after mastication. Mincing reproduced the division of bread and pasta as achieved by chewing in an acceptable way. From our results it seems that the size of particles resulting from mastication depends on food texture. We succeeded by wetting and mincing food to prepare food in a similar bolus-like form before swallowing. Mincing provides a simple means of simulating the reduction of food into particles for in vitro digestion studies.

Non-starch polysaccharides extracted from seaweed can modulate intestinal absorption of glucose and insulin response in the pig.

We have investigated the possible effects of algal polysaccharides on postprandial blood glucose and insulin responses in an animal model, the pig. Three seaweed fibres of different viscosities, extracted from Palmaria palmata (PP), Eucheuma cottonii (EC), or Laminaria digitata (LD), were compared to purified cellulose (CEL). Blood glucose and plasma insulin levels were monitored and intestinal absorption quantified for 8 h following a high carbohydrate test-meal supplemented with 5% fibre. Digestive contents were also sampled, 5 h postprandial. As compared to CEL, PP had no effect on glucose and insulin responses. The latter decreased with EC, but glucose absorption balance was not modified. LD addition resulted in a dramatically reduced glucose absorption balance, accompanied by a higher amount of starch left in the small intestine. Among polysaccharides tested, only the highly viscous alginates could affect intestinal absorption of glucose and insulin response.

Bioavailability of starch in bread rich in amylose: metabolic responses in healthy subjects and starch structure.

OBJECTIVE: This study investigated whether postprandial metabolic responses to bread could be lowered by substituting high amylose maize starch for a part of the flour. DESIGN AND SUBJECTS: Eight healthy subjects consumed test meals of equivalent nutritional composition based on white wheat bread, bread rich in amylose (HAWB) and spaghetti as a breakfast meal. Blood samples were collected to measure insulin and glucose concentration during two hours after consumption. The degree of starch crystallinity was investigated by X-ray diffraction and DSC analysis. RESULTS: HAWB produced low glycaemic (60 +/- 18) and insulinaemic (57 +/- 20) indexes similar to those of spaghetti (83 +/- 46, 61 +/- 16). In vitro amylase hydrolysis of the three foods showed that high amylose content in HAWB significantly lowered starch degradation in bread without affecting hydrolysis kinetics. Addition of amylose in dough increased the resistant starch content of HAWB (14% of dry matter). The resistant starch fraction was mainly composed of crystalline amylose (B-type X-ray diffraction pattern, melting temperature 105 degrees C) attributable to native high amylose maize starch incompletely gelatinised during bread-cooking. CONCLUSIONS: Bread produced by the substitution of high amylose maize starch for a part of wheat flour showed a low glycaemic index. Resistant starch in HAWB corresponded to native crystalline amylose not gelatinised during normal bread-processing conditions.

Physical and chemical transformations of cereal food during oral digestion in human subjects.

Chemical and physical transformations of solid food begin in the mouth, but the oral phase of digestion has rarely been studied. In the present study, twelve healthy volunteers masticated mouthfuls of either bread or spaghetti for a physiologically-determined time, and the levels of particle degradation and starch digestion before swallowing were compared for each food. The amounts of saliva moistening bread and spaghetti before swallowing were, respectively, 220 (SEM 12) v. 39 (SEM 6) g/kg fresh matter. Particle size reduction also differed since bread particles were highly degraded, showing a loss of structure, whereas spaghetti retained its physical structure, with rough and incomplete reduction of particle size. Starch hydrolysis was twice as high for bread as for spaghetti, mainly because of the release of high-molecular-mass alpha-glucans. The production of oligosaccharides was similar after mastication of the two foods, respectively 125 (SEM 8) and 92 (SEM 7) g/kg total starch. Starch hydrolysis, which clearly began in the mouth, depended on the initial structure of the food, as in the breakdown of solid food. These significant physical and chemical degradations of solid foods during oral digestion may influence the entire digestive process.

Estimation of the fermentability of dietary fibre in vitro: a European interlaboratory study.

Five European laboratories tested a simple in vitro batch system for dietary fibre fermentation studies. The inoculum was composed of fresh human faeces mixed with a carbonate-phosphate buffer complex supplemented with trace elements and urea. Five dietary fibre sources (cellulose, sugarbeet fibre, soyabean fibre, maize bran and pectin) were used by each laboratory on three occasions to determine pH, residual non-starch polysaccharides (NSP) and short-chain fatty acid production during fermentation. Cellulose and maize bran degradabilities were very low (7.2(SE 10.8) and 6.2 (SE 9.1)% respectively after 24 h), whereas pectin and soyabean fibre were highly degraded (97.4 (SE 4.4) and 91.1 (SE 3.4)% respectively after 24 h). Sugarbeet fibre exhibited an intermediate level of degradability (59.5 (SE 14.9)%). Short-chain fatty acid production was closely related to NSP degradation (r 0.99). Although each variable was ranked similarly by all laboratories, some differences occurred with respect to absolute values. However, the adaptation of donors to the experimental substrates was not an influential factor. Interlaboratory differences could be reduced either by adding less substrate during incubations or using less-diluted inocula. In vitro fermentations with inocula made from human faeces and from rat caecal contents gave similar results. There was a close correspondence between the data obtained in the present experiment and those previously published in in vivo studies in the rat using the same fibres. The in vitro batch system tested during the present study provides a rapid means of obtaining quantitative estimates of the fermentation and the estimation of the energy content of new sources of dietary fibre.

[Rapid assay of plasma acetate by gas chromatography: evaluation and comparison with two other methods]. / Dosage rapide de l'acétate plasmatique par chromatographie en phase gazeuse--technique espace de tête: évaluation et comparaison avec deux autres méthodes.

Human plasma acetate is derived from colonic fermentation of fiber and endogenous metabolism of dextrose and fatty acids. Acetate may have regulatory functions in hepatic carbohydrate metabolism. Intake of dietary fiber is associated with several beneficial effects on carbohydrates and lipids metabolisms. To study theses effects a valid and automated method for routine analysis of acetate in plasma is necessary. After oral administration of lactulose to healthy human volunteers, the concentration of plasma acetate was measured by head space gas chromatography (HS-GC), vacuum distillation gas chromatography (VD-GC) and enzymatic spectrometric method (ES). The method HS-GC was linear to 0.5 mmol.l-1 (n = 5, r = 0.998), the detection limit is 0.005 mmol.l-1. Within-day variation (CV) was 3.60% and day-to-day variation was 4.5% (0.1 mmol.l-1). The coefficients of correlation between CG-ET/CG-DsV and CG-ET/E-M are 0.903 (p = 0.0001) and 0.54 (p = 0.006) respectively, the mean square errors are respectively 0.118 and 0.138 mmol.l-1. The variation curves of plasma acetate measured by GC versus time show peak concentration of 0.323 to 0.380 mmol.l-1 at 120 min.