Inter-laboratory validation of the starch digestibility method for determination of rapidly digestible and slowly digestible starch.

The digestibility of starch in foods, which is influenced by the ingredients, formulation and preparation conditions, is a major determinant of glycaemic response. The terms rapidly digestible starch (RDS) and slowly digestible starch (SDS) along with the associated analytical methodology were developed by Englyst to characterise this nutritionally relevant food attribute. The measurement uncertainty of this starch digestibility method is evaluated here with an inter-laboratory trial. Six laboratories took part in the study testing ten cereal products with mean (range) contents of RDS: 48.4 g/100 g, (23.4-76·9) and, SDS: 10.9 g/100 g, (0.8-24.2). Based on the repeatability and reproducibility measurements, the calculated uncertainty was 3.6 g/100 g for RDS and 1.9 g/100 g for SDS. This trial has demonstrated acceptable measurement uncertainty and confirmed the transferability of the method between laboratories. The SDS content can identify foods rich in slow release carbohydrates with their associated health benefits.

Evaluation of methods of analysis for dietary fibre using real foods and model foods.

RATIONALE AND OBJECTIVES: There are several rational and empirical methods for the measurement of dietary fibre and its components. A selection of these methods were evaluated by investigation of a range of real foods and model foods with added resistant starch (RS), non-starch polysaccharides (NSP) and resistant oligosaccharide (RO) ingredients. METHODS: A range of rational methods were applied in determining specific carbohydrate constituents: RS, NSP and RO, including fructans. For comparison, empirical methods AOAC 991.43 (2001.03) and AOAC 2009.01 were applied, based on determination of gravimetric residues for high molecular weight and size-exclusion HPLC analysis of the ethanol filtrate for low molecular weight components. RESULTS: In general there was agreement between different rational methods for the analysis of RS and fructans, though there were notable exceptions for some product types. Comparison of methods for total RS and those that only measure the RS3 fraction, from retrograded starch, indicated that RS3 was the only type present for most processed products. This also explains the similar results obtained by AOAC 991.43 (2001.03) and AOAC 2009.01, though the latter is intended to recover other RS types as well. For many products there was agreement between results obtained by rational and empirical methods, though there were exceptions and the reasons for these are discussed. CONCLUSION: Rational and empirical methods can both be used to determine dietary fibre in most situations. The information provided by rational methods is useful in identifying the specific carbohydrate constituents present in foods and can assist in determining whether added extracted and synthesised ingredients are ones that conform to the Codex and EU dietary fibre definition.

Resistant carbohydrates stimulate cell proliferation and crypt fission in wild-type mice and in the Apc(Min/+) mouse model of intestinal cancer, association with enhanced polyp development.

Fermentation of carbohydrates in the colon can stimulate cell proliferation and could thus be a cancer risk. The effects of resistant carbohydrates, i.e. those not digested and absorbed in the small intestine, on cell proliferation, crypt fission and polyp development were investigated in wild-type and adenomatous polyposis coli multiple intestinal neoplasia (Apc(Min/+)) mice. Fifteen 4-week-old female wild-type and fifteen Apc(Min/+) mice were used for each group and fed a chow diet, a semi-synthetic diet or the semi-synthetic supplemented with wheat bran or an apple pomace preparation, both high in resistant carbohydrates, for 8 weeks. Tissue from all mice was used to measure cell proliferation and crypt fission and tissue from the Apc(Min/+) mice was scored for polyp number and tumour burden. There were slight reductions in intestinal mass in the mice fed the semi-synthetic diets and this was increased by the inclusion of resistant carbohydrates. The Apc(Min/+) mice had elevated cell proliferation and crypt fission in the distal small intestine and colon and these were increased by the resistant carbohydrates. Bran or apple pomace significantly increased polyp number in the proximal third of the small intestine. Apple pulp more than doubled polyp number throughout the small bowel (99.2 (SEM 11.1) v. 40.0 (SEM 8.2), P<0.004). Bran and apple pomace increased polyp diameter and hence burden in the colon by 243 and 150 %, respectively (P<0.05). In conclusion, both types of resistant carbohydrates increased polyp number and tumour burden and this was associated with elevated epithelial cell proliferation and crypt fission.

Carbohydrate bioavailability.

There is consensus that carbohydrate foods, in the form of fruit, vegetables and whole-grain products, are beneficial to health. However, there are strong indications that highly processed, fibre-depleted, and consequently rapidly digestible, energy-dense carbohydrate food products can lead to over-consumption and obesity-related diseases. Greater attention needs to be given to carbohydrate bioavailability, which is determined by the chemical identity and physical form of food. The objective of the present concept article is to provide a rational basis for the nutritional characterisation of dietary carbohydrates. Based on the properties of carbohydrate foods identified to be of specific relevance to health, we propose a classification and measurement scheme that divides dietary carbohydrates into glycaemic carbohydrates (digested and absorbed in the small intestine) and non-glycaemic carbohydrates (enter the large intestine). The glycaemic carbohydrates are characterised by sugar type, and by the likely rate of digestion described by in vitro measurements for rapidly available glucose and slowly available glucose. The main type of non-glycaemic carbohydrates is the plant cell-wall NSP, which is a marker of the natural fibre-rich diet recognised as beneficial to health. Other non-glycaemic carbohydrates include resistant starch and the resistant short-chain carbohydrates (non-digestible oligosaccharides), which should be measured and researched in their own right. The proposed classification and measurement scheme is complementary to the dietary fibre and glycaemic index concepts in the promotion of healthy diets with low energy density required for combating obesity-related diseases.

Enhanced Escherichia coli adherence and invasion in Crohn's disease and colon cancer.

BACKGROUND & AIMS: Altered mucosal glycosylation in inflammatory bowel disease and colon cancer could affect mucosal bacterial adherence. This study aimed to quantify and characterize mucosa-associated and intramucosal bacteria, particularly Escherichia coli, in these conditions. METHODS: Mucosa-associated bacteria were isolated, after dithiothreitol mucolysis, from biopsy samples obtained at colonoscopy (Crohn's disease, n = 14 patients; ulcerative colitis, n = 21; noninflamed controls, n = 24) and at surgical resection (colon cancer, n = 21). Intramucosal bacteria were grown after gentamicin treatment followed by hypotonic lysis. RESULTS: Mucosa-associated and intramucosal bacteria were cultured more commonly in Crohn's disease (79%, P = 0.03; and 71%, P < 0.01, respectively), but not ulcerative colitis (38% and 48%), than in noninflamed controls (42% and 29%) and were commonly cultured from colon cancers (71% and 57%). Mucosa-associated E. coli, which accounted for 53% of isolates, were more common in Crohn's disease (6/14; 43%) than in noninflamed controls (4/24, 17%), as also were intramucosal E. coli: Crohn's disease, 29%; controls, 9%. E. coli expressed hemagglutinins in 39% of Crohn's cases and 38% of cancers but only 4% of controls, and this correlated (P = 0.01) with adherence to the I407 and HT29 cell lines. Invasion was cell-line dependent. E. coli, including nonadherent isolates, induced interleukin-8 release from the cell lines. E. coli adhesins showed no blood group specificity, excepting 1 cancer isolate (HM44) with specificity for the Thomsen-Friedenreich antigen, but they could be blocked by soluble plantain fiber. CONCLUSIONS: These studies support a central role for mucosally adherent bacteria in the pathogenesis of Crohn's disease and colon cancer. Soluble plant fibers that inhibit their adherence have therapeutic potential.

Degradation of cross-linked and non-cross-linked arabinoxylans by the intestinal microbiota in children.

In humans, nonstarch polysaccharides (NSP), such as arabinoxylans (AX), are not digested in the upper gut and provide fermentable carbon sources for bacteria growing in the large bowel. Despite the ubiquity of AX in nature, the microbiologic and physiologic consequences of AX digestion in the gut are poorly understood. In this study, we investigated the breakdown of ferulic acid-cross-linked AX (AXF) and non-cross-linked AX in children's intestinal microbiotas, using starch as a readily fermentable polysaccharide for comparative purposes. The experiments were performed using pH-controlled fermentation vessels under anaerobic conditions. The results demonstrated that there was variation in the metabolism of these polysaccharides by colonic microbiotas. AX was always degraded more slowly than starch, while ferulic acid cross-linking reduced the rate of AX fermentation, as shown by fermentation product measurements. Starch digestion was associated with significant acetate and butyrate production, whereas AX breakdown resulted in increased propionate formation. In general, the presence of fermentable carbohydrate significantly increased the total anaerobe counts and eubacterial rRNA concentrations (P < 0.01), while non-cross-linked AX digestion was principally associated with increased viable counts of Bacteroides fragilis group organisms, which was supported by increases in Bacteroides-Porphyromonas-Prevotella group rRNA (P < 0.01). Starch was considerably more bifidogenic than AX in these fermentations. In conclusion, in this study we found that the effects of AX and AXF on the microbial ecology and metabolism of intestinal microbiotas are similar in children and adults.

Glycaemic index of cereal products explained by their content of rapidly and slowly available glucose.

Elucidating the role of carbohydrate quality in human nutrition requires a greater understanding of how the physico-chemical characteristics of foods relate to their physiological properties. It was hypothesised that rapidly available glucose (RAG) and slowly available glucose (SAG), in vitro measures describing the rate of glucose release from foods, are the main determinants of glycaemic index (GI) and insulinaemic index (II) for cereal products. Twenty-three products (five breakfast cereals, six bakery products and crackers, and twelve biscuits) had their GI and II values determined, and were characterised by their fat, protein, starch and sugar contents, with the carbohydrate fraction further divided into total fructose, RAG, SAG and resistant starch. Relationships between these characteristics and GI and II values were investigated by regression analysis. The cereal products had a range of GI (28-93) and II (61-115) values, which were positively correlated (r(2)) 0.22, P<0.001). The biscuit group, which had the highest SAG content (8.6 (SD 3.7) g per portion) due to the presence of ungelatinised starch, was found to have the lowest GI value (51 (SD 14)). There was no significant association between GI and either starch or sugar, while RAG was positively (r(2)) 0.54, P<0.001) and SAG was negatively (r(2)) 0.63, P<0.001) correlated with GI. Fat was correlated with GI (r(2)) 0.52, P<0.001), and combined SAG and fat accounted for 73.1% of the variance in GI, with SAG as the dominant variable. RAG and protein together contributed equally in accounting for 45.0 % of the variance in II. In conclusion, the GI and II values of the cereal products investigated can be explained by the RAG and SAG contents. A high SAG content identifies low-GI foods that are rich in slowly released carbohydrates for which health benefits have been proposed.

The importance of dietary carbohydrates.

Forty years ago carbohydrates (CHO) were regarded as a simple energy source whereas they are now recognized as important food components. The human diet contains a wide range of CHO, the vast majority of which are of plant origin. Modern techniques based on chemical classification of dietary CHO replaced the traditional "by difference" measurement. They provide a logical basis for grouping into categories of specific nutritional importance. The physiological effects of dietary CHO are highly dependent on the rate and extent of digestion and absorption in the small intestine and fermentation in the large intestine, interactions which promote human health. Current knowledge of the fate of dietary CHO means that the potentially undesirable properties of many modern foods could be altered by using processing techniques that yield foods with more intact plant cell wall structures. Such products would more closely resemble the foods in the pre-agriculture diet with respect to the rate of digestion and absorption of CHO in the small intestine. The potentially detrimental physiological consequences of eating sugars and starch that are rapidly digested and absorbed in the small intestine suggest that, as fibre, the form, as well as the amount of starch should be considered. Increasing consumer awareness of the relationship between diet and health has led to demands for more widespread nutrition labelling. The entry "carbohydrate" is required in most countries, and the value is usually obtained "by difference" and used in the calculation of energy content. However, the value provides no nutritional information per se. Food labels should provide values that aid consumers in selecting a healthy diet.