Substitution of Refined Conventional Wheat Flour with Wheat High in Resistant Starch Modulates the Intestinal Microbiota and Fecal Metabolites in Healthy Adults: A Randomized, Controlled Trial.

BACKGROUND: Resistant starch (RS) confers many health benefits, mostly through the microbial production of SCFAs, but foods containing appreciable RS are limited. High-amylose wheat (HAW) is high in RS and lowers the glycemic response of foods, but whether it can improve gastrointestinal health measures is unknown. OBJECTIVES: The objective of this study was to determine whether daily consumption of HAW food products improved markers of gastrointestinal health in healthy men and women compared with similar foods made from conventional wheat. METHODS: Eighty healthy adults (47 women and 33 men) were enrolled in a 4-arm parallel, randomized-controlled, double-blind trial. After a 2-wk low-dietary fiber run-in period, they were randomly allocated to 1 of 4 treatment groups: low-amylose wheat (LAW)-refined (LAW-R), LAW-wholemeal (LAW-W), HAW-refined (HAW-R), and HAW-wholemeal (HAW-W) and consumed the assigned test bread (160 g/d) and biscuits (75 g/d) for 4 wk. Fecal biochemical markers were measured at baseline and 4 wk. Microbial abundance and diversity were quantified using 16S ribosomal RNA sequencing and perceived gut comfort by a semiquantitative questionnaire completed at baseline, 2 wk, and 4 wk. RESULTS: HAW showed similar effects on fecal output and excretion of total SCFA compared with LAW, but changes were observed in secondary measures for the refined treatment groups. At 4 wk, the HAW-R group had 38% higher fecal butyrate excretion than the LAW-R group (P < 0.05), and higher fecal SCFA-producing bacteria, Roseburia inulinivorans (P < 0.001), than at baseline. In comparison with baseline, LAW-R increased fecal p-cresol concentration, and fecal abundance of a p-cresol-producing bacterium, Clostridium from the Peptostreptococcaceae family, but both were reduced by HAW-R. Amylose level did not affect measures of fecal consistency or adversely affecting digestive comfort. CONCLUSIONS: Increasing RS intake of healthy adults by substituting refined conventional wheat with refined HAW modulates fecal metabolites and microbes associated with gastrointestinal health.This trial was registered at anzctr.org.au as ACTRN12618001060235.

Rice with Multilayer Aleurone: A Larger Sink for Multiple Micronutrients.

Diet-related noncommunicable diseases impose a heavy burden on human health worldwide. Rice is a good target for diet-related disease prevention strategies because it is widely consumed. Liu et al. (Proc Natl Acad Sci USA 115(44):11327-11332, 2018. https://doi.org/10.1073/pnas.1806304115 ) demonstrated that increasing the number of cell layers and thickness of putative aleurone in ta2-1 (thick aleurone 2-1) mutant rice enhances simultaneously the content of multiple micronutrients. However, the increases of aleurone-associated nutrients were not proportional to the increases in the aleurone thickness. In this study, first, cytohistological analyses and transmission electron microscopy demonstrated that the multilayer in ta2-1 exhibited aleurone cell structural features. Second, we detected an increase in insoluble fibre and insoluble bound-phenolic compounds, a shift in aleurone-specific neutral non-starch polysaccharide profile, enhancement of phytate and minerals such as iron, zinc, potassium, magnesium, sulphur, and manganese, enrichment of triacylglycerol and phosphatidylcholine but slight reduction in free fatty acid, and an increase in oleic fatty acid composition. These findings support our hypothesis that the expanded aleurone-like layers in ta2-1 maintained some of the distinctive aleurone features and composition. We provide perspectives to achieve even greater filling of this expanded micronutrient sink to provide a means for multiple micronutrient enhancements in rice.

Functional Genomic Validation of the Roles of Soluble Starch Synthase IIa in Japonica Rice Endosperm.

The enzyme starch synthase IIa (SSIIa) in cereals has catalytic and regulatory roles during the synthesis of amylopectin that influences the functional properties of the grain. Rice endosperm SSIIa is more active in indica accessions compared to japonica lines due to functional SNP variations in the coding region of the structural gene. In this study, downregulating the expression of japonica-type SSIIa in Nipponbare endosperm resulted in either shrunken or opaque grains with an elevated proportion of A-type starch granules. Shrunken seeds had severely reduced starch content and could not be maintained in succeeding generations. In comparison, the opaque grain morphology was the result of weaker down-regulation of SSIIa which led to an elevated proportion of short-chain amylopectin (DP 6-12) and a concomitant reduction in the proportion of medium-chain amylopectin (DP 13-36). The peak gelatinization temperature of starch and the estimated glycemic score of cooked grain as measured by the starch hydrolysis index were significantly reduced. These results highlight the important role of medium-chain amylopectin in influencing the functional properties of rice grains, including its digestibility. The structural, regulatory and nutritional implications of down-regulated japonica-type SSIIa in rice endosperm are discussed.

Evaluation of an Ileorectostomised Rat Model for Resistant Starch Determination.

The human ileostomy model, widely considered the benchmark for determining in vivo starch digestibility, has disadvantages. The ileorectostomised rat model (IRM) is a possible surrogate but evidence as to its validity is scant. In this preliminary study, the resistant starch (RS) content of test breads made from refined low (LAW-R) and high amylose wheat (HAW-R) flours was established in a randomised cross-over trial involving six human ileostomy participants. Starch digestibility of refined breads and diets made from these flours was then evaluated in ileorectostomised rats using a similar experimental format. Physical performance measures and other data were also collected for the rat model. The amount of RS in the low- and high-amylose breads as measured using the human model was 0.8 ± 0.1 and 6.5 ± 0.3 g/100 g, respectively. The RS level of HAW-R bread determined using ileorectostomised rats was 5.5 ± 0.8 g/100 g, about 15% less than that recorded in the human study, whereas for conventional wheat breads the models produced similar RS values. While offering promise, further validation using a wide variety of starchy food products is needed before the IRM can be considered an acceptable alternative for RS determination.

Long-term Paleolithic diet is associated with lower resistant starch intake, different gut microbiota composition and increased serum TMAO concentrations.

BACKGROUND: The Paleolithic diet is promoted worldwide for improved gut health. However, there is little evidence available to support these claims, with existing literature examining anthropometric and cardiometabolic outcomes. OBJECTIVE: To determine the association between dietary intake, markers of colonic health, microbiota, and serum trimethylamine-N-oxide (TMAO), a gut-derived metabolite associated with cardiovascular disease. DESIGN: In a cross-sectional design, long-term (n = 44, > 1 year) self-reported followers of a Paleolithic diet (PD) and controls (n = 47) consuming a diet typical of national recommendations were recruited. Diets were assessed via 3-day weighed diet records; 48-h stool for short chain fatty acids using GC/MS, microbial composition via 16S rRNA sequencing of the V4 region using Illumina MiSeq. TMAO was quantified using LC-MS/MS. RESULTS: Participants were grouped according to PD adherence; namely excluding grains and dairy products. Strict Paleolithic (SP) (n = 22) and Pseudo-Paleolithic (PP) (n = 22) groups were formed. General linear modelling with age, gender, energy intake and body fat percentage as covariates assessed differences between groups. Intake of resistant starch was lower in both Paleolithic groups, compared to controls [2.62, 1.26 vs 4.48 g/day (P < 0.05)]; PERMANOVA analysis showed differences in microbiota composition (P < 0.05), with higher abundance of TMA-producer Hungatella in both Paleolithic groups (P < 0.001). TMAO was higher in SP compared to PP and control (P < 0.01), and inversely associated with whole grain intake (r = - 0.34, P < 0.01). CONCLUSIONS: Although the PD is promoted for improved gut health, results indicate long-term adherence is associated with different gut microbiota and increased TMAO. A variety of fiber components, including whole grain sources may be required to maintain gut and cardiovascular health. CLINICAL TRIAL REGISTRATIONS: Australian and New Zealand Clinical Trial Registry (ANZCTRN12616001703493).

High-Amylose Wheat Lowers the Postprandial Glycemic Response to Bread in Healthy Adults: A Randomized Controlled Crossover Trial.

BACKGROUND: Conventional wheat-based foods contain high concentrations of readily digestible starch that commonly give these foods a high postprandial glycemic response and may contribute to the development of type 2 diabetes and cardiovascular disease. OBJECTIVES: The aim of this study was to determine if bread made from high-amylose wheat (HAW) and enriched in resistant starch dampens postprandial glycemia compared with bread made from conventional low-amylose wheat (LAW). METHODS: This single-center, randomized, double-blinded, crossover controlled study involved 7 consecutive weekly visits. On separate mornings, 20 healthy nondiabetic men and women (mean age 30 ± 3 y; body mass index 23 ± 0.7 kg/m2) consumed a glucose beverage or 4 different breads (each 121 g); LAW-R (refined), LAW-W (wholemeal), HAW-R, or HAW-W. The starch contents of the LAW and HAW breads were 24% and 74% amylose, respectively. Venous blood samples were collected at regular intervals before and for 3 h after the breakfast meal to measure plasma glucose, insulin, ghrelin, and incretin hormone concentrations, and the incremental area under the curve (AUC) was calculated (mmol/L × 3 h). Satiety and cravings were also measured at 30-min intervals during the postprandial period. RESULTS: HAW breads had a glycemic response (AUC) that was 39% less than that achieved with conventional wheat breads (HAW 39 ± 5 mmol/L × 3 h; LAW 64 ± 5 mmol/L × 3 h; P < 0.0001). Insulinemic and incretin responses were 24-30% less for HAW breads than for LAW breads (P < 0.05). Processing of the flour (wholemeal or refined) did not affect the glycemic, insulinemic, or incretin response. The HAW breads did not influence plasma ghrelin, or subjective measures of satiety or cravings during the postprandial period. CONCLUSIONS: Replacing LAW with HAW flour may be an effective strategy for lowering postprandial glycemic and insulinemic responses to bread in healthy men and women, but further research is warranted. This trial was registered at the Australian and New Zealand Clinical Trials Registry as ACTRN12616001289404.

Resistant Starch is Actively Fermented by Infant Faecal Microbiota and Increases Microbial Diversity.

In adults, fermentation of high amylose maize starch (HAMS), a resistant starch (RS), has a prebiotic effect. Were such a capacity to exist in infants, intake of RS might programme the gut microbiota during a critical developmental period. This study aimed to determine if infant faecal inocula possess the capacity to ferment HAMS or acetylated-HAMS (HAMSA) and characterise associated changes to microbial composition. Faecal samples were collected from 17 healthy infants at two timepoints: Preweaning and within 10 weeks of first solids. Fermentation was assessed using in vitro batch fermentation. Following 24 h incubation, pH, short-chain fatty acid (SCFA) production and microbial composition were compared to parallel control incubations. In preweaning infants, there was a significant decrease at 24 h in pH between control and HAMS incubations and a significant increase in the production of total SCFAs, indicating fermentation. Fermentation of HAMS increased further following commencement of solids. Fermentation of RS with weaning faecal inocula increased Shannon's diversity index (H) and was associated with increased abundance of Bifidobacterium and Bacteroides. In conclusion, the faecal inocula from infants is capable of RS fermentation, independent of stage of weaning, but introduction of solids increases this fermentation capacity. RS may thus function as a novel infant prebiotic.

Cereal fructan extracts alter intestinal fermentation to reduce adiposity and increase mineral retention compared to oligofructose.

PURPOSE: Intestinal fermentation of inulin-type fructans, including oligofructose, can modulate adiposity, improve energy regulation, and increase mineral absorption. We aimed to determine whether cereal fructans had greater effects on reducing adiposity and improving mineral absorption compared with oligofructose. METHODS: Thirty-two male Sprague-Dawley rats were randomly assigned to one of four dietary treatments that contained 0% fructan (control), or 5% fructan provided by oligofructose (OF), a barley grain fraction (BGF), or a wheat stem fraction (WSF). After 1 week on the diets, mineral absorption and retention was assessed. At 4 weeks, blood samples were collected for gut hormone analysis, adipose depots were removed and weighed, and caecal digesta was analyzed for pH and short-chain fatty acids (SCFA). RESULTS: The BGF and WSF, but not OF, had lower total visceral fat weights than the Control (p < 0.05). The fructan diets all lowered caecal pH and raised caecal digesta weight and total SCFA content, in comparison to the Control. Caecal propionate levels for OF were similar to the Control and higher for WSF (p < 0.05). Plasma peptide YY and glucagon-like peptide-1 levels were elevated for all fructan groups when compared to Control (p < 0.001) and gastric inhibitory peptide was lower for the WSF compared to the other groups (p < 0.05). The fructan diets improved calcium and magnesium retention, which was highest for WSF (p < 0.05). BGF and WSF in comparison to OF showed differential effects on fermentation, gut hormone levels, and adiposity. CONCLUSIONS: Cereal fructan sources have favorable metabolic effects that suggest greater improvements in energy regulation and mineral status to those reported for oligofructose.

Integrating a genome-wide association study with a large-scale transcriptome analysis to predict genetic regions influencing the glycaemic index and texture in rice.

Reliably generating rice varieties with low glycaemic index (GI) is an important nutritional intervention given the high rates of Type II diabetes incidences in Asia where rice is staple diet. We integrated a genome-wide association study (GWAS) with a transcriptome-wide association study (TWAS) to determine the genetic basis of the GI in rice. GWAS utilized 305 re-sequenced diverse indica panel comprising ~2.4 million single nucleotide polymorphisms (SNPs) enriched in genic regions. A novel association signal was detected at a synonymous SNP in exon 2 of LOC_Os05g03600 for intermediate-to-high GI phenotypic variation. Another major hotspot region was predicted for contributing intermediate-to-high GI variation, involves 26 genes on chromosome 6 (GI6.1). These set of genes included GBSSI, two hydrolase genes, genes involved in signalling and chromatin modification. The TWAS and methylome sequencing data revealed cis-acting functionally relevant genetic variants with differential methylation patterns in the hot spot GI6.1 region, narrowing the target to 13 genes. Conversely, the promoter region of GBSSI and its alternative splicing allele (G allele of Wxa ) explained the intermediate-to-high GI variation. A SNP (C˃T) at exon-10 was also highlighted in the preceding analyses to influence final viscosity (FV), which is independent of amylose content/GI. The low GI line with GC haplotype confirmed soft texture, while other two low GI lines with GT haplotype were characterized as hard and cohesive. The low GI lines were further confirmed through clinical in vivo studies. Gene regulatory network analysis highlighted the role of the non-starch polysaccharide pathway in lowering GI.

Investigating glycemic potential of rice by unraveling compositional variations in mature grain and starch mobilization patterns during seed germination.

Rice lines with slower starch digestibility provide opportunities in mitigating the global rise in type II diabetes and related non-communicable diseases. However, screening for low glycemic index (GI) in rice breeding programs is not possible due to time and cost constraints. This study evaluated the feasibility of using in vitro cooked grain amylolysis, starch mobilization patterns during seed germination, and variation in starch structure and composition in the mature seed to differentiate patterns of starch digestibility. Mobilization patterns of total starch, resistant starch, amylose and amylopectin chains, and free sugars during seed germination revealed that the process is analogous to digestion in the human gastrointestinal tract. The combination of these biochemical markers can be used as an alternative measure to predict GI. Additionally, transcriptome analysis of stored mRNA transcripts in high and low GI lines detected differences in starch metabolism and confirmed the importance of seed storage pathways in influencing digestibility. Pathway analyses supported by metabolomics data revealed that resistant starch, cell wall non-starch polysaccharides and flavonoids potentially contribute to slower digestibility. These new insights can guide precision breeding programs to produce low GI rice with acceptable cooking quality to help mitigate the burden of diet-associated lifestyle diseases.

A rat model for determining the postprandial response to foods.

BACKGROUND: The use of small animal models for studying postprandial changes in circulating nutrients, hormones and metabolic biomarkers is hampered by the limited quantity of blood that can be withdrawn for analysis. Here, we describe the development of an unrestrained, meal-fed rat model, having a permanent or temporary vascular cannula that permits repeated blood sampling. The applicability and performance of the model were evaluated in a series of experiments on acute glycaemic and insulinaemic responses to carbohydrate-based test meals. RESULTS: A test food containing 0.4 g carbohydrate raised blood glucose by 1.5 mmol L-1 . Postprandial blood glucose levels peaked at 15 min and returned to baseline at 180 min, whereas they remained elevated for longer when the test meal contained 1.25 g carbohydrate. The glycaemic response tended (P = 0.063) to be higher when the meal tolerance test was conducted at the start rather than the end of the dark period, but the insulinaemic response was unaffected. The magnitude of the glycaemic response was less for blood collected from the caudal vein compared to that from the jugular vein. Both cannulation strategies were equally effective in enabling return of red blood cells, thus preserving blood volume. CONCLUSION: This improved small animal model affords new opportunities to screen foods for nutrient bioavailability and explore metabolic mechanisms mediating responses to food consumption. © 2016 Society of Chemical Industry.

Microencapsulated krill and tuna oil blend raises plasma long-chain n-3 polyunsaturated fatty acid levels compared to tuna oil with similar increases in ileal contractility in rats.

Long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) may be more bioavailable from krill oil compared to fish oil due to their phospholipid structure. We tested whether a microencapsulated krill and tuna oil blend (ME-TOKO) provided greater LC n-3 PUFA bioavailability, improved blood lipid profiles and increased intestinal contractility compared to microencapsulated tuna oil (ME-TO). Rats were divided into three groups to receive isocaloric diets containing ME-TO, ME-TOKO and microencapsulated olive oil (ME-OO) at 0.3 or 2 g/100 g for 4 weeks. Final body and organ weights, feed intake and waste output were similar. ME-TOKO rats had higher plasma total LC n-3 PUFA levels compared to ME-TO, but liver LC n-3 PUFA levels and plasma triglyceride and cholesterol levels were similar in non-fasted rats. Diets containing 2% ME-TO and ME-TOKO also showed similar increases in ileal contractility. In summary, ME-TO bioavailability of LC n-3 PUFA was similar to ME-TOKO.

High wholegrain barley ß-glucan lowers food intake but does not alter small intestinal macronutrient digestibility in ileorectostomised rats.

Using barley cultivars differing widely in ß-glucan content, we aimed to determine their effects on small intestinal macronutrient digestion in 24 ileorectostomised rats. The rats were fed 1 of 4 experimental diets, each containing a different barley variety, for 11 d. The diets had a content of 0, 2.1, 2.6 and 4.3 g of ß-glucan/100 g. Feed intake and faecal excretion of fat, protein, starch, and non-starch polysaccharides were determined in the final 5 d of the study and apparent macronutrient digestibility calculated. Higher dietary levels of ß-glucan (2.6% and 4.3%) lowered feed intake (by 15 and 19%, respectively) but final body weight was only lowered by the 4.3% ß-glucan diet relative to rats fed the 0% ß-glucan diet (all ps < 0.05). Protein, lipid and starch digestibility was unrelated to the dietary ß-glucan content. Higher dietary levels of barley ß-glucan lower feed intake of ileorectostomised rats, which is independent of intestinal fermentation and unrelated to macronutrient digestibility.

Consistent Prebiotic Effect on Gut Microbiota With Altered FODMAP Intake in Patients with Crohn's Disease: A Randomised, Controlled Cross-Over Trial of Well-Defined Diets.

OBJECTIVES: Altering FODMAP (fermentable oligosaccharides, disaccharides, monosaccharides and polyols) intake has substantial effects on gut microbiota. This study aimed to investigate effects of altering FODMAP intake on markers of colonic health in patients with Crohn's disease. METHODS: After evaluation of their habitual diet, 9 patients with clinically quiescent Crohn's disease were randomised to 21 days of provided low or typical ("Australian") FODMAP diets with ≥21-day washout in between. Five-day fecal samples were collected at the end of each diet and analyzed for calprotectin, pH, short-chain fatty acids (SCFA) and bacterial abundance. Gastrointestinal symptoms were recorded daily. RESULTS: Eight participants collected feces and were adherent to the diets. FODMAP intake differed across the three dietary periods with low

Circulating triglycerides and bile acids are reduced by a soluble wheat arabinoxylan via modulation of bile concentration and lipid digestion rates in a pig model.

SCOPE: Soluble dietary fibres have shown to have lipid reducing properties. However, their mechanisms of action are still unclear. The present study investigated how a soluble wheat arabinoxylan-rich fraction (AXRF) fed to pigs used as a human model reduced blood triglycerides. METHODS AND RESULTS: After 4 weeks on the experimental diets, blood from the jugular (JV) and hepatic portal (HPV) veins, bile from the gall bladder, and digesta samples from four sites of the small intestine (SI) and cecum were collected. The results showed that the AXRF significantly decreased the concentrations of total bile acid (BA) in the HPV (p < 0.01), JV (p < 0.01), bile (p < 0.05) and SI (p < 0.05), but with no effect on ileal BAs excretion flux. Furthermore, blood triglyceride (TAG) levels were also lower with AXRF (p < 0.01) but with no significant effects on LDL-, HDL- or total cholesterol levels. The lower plasma TAG concentration was consistent with the reduced/delayed digestion and absorption of TAG with the AXRF (total fatty acid and MUFA p < 0.01; unsaturated fatty acid p < 0.05). CONCLUSION: The results suggest that AXRF reduced the levels of circulating BAs which slowed down the digestion of TAG and absorption of free fatty acids, with consequent reduction in blood TAG. Reduction in circulating bile acids by arabinoxylan causes reduction in lipids digestion and absorption.

Soluble arabinoxylan alters digesta flow and protein digestion of red meat-containing diets in pigs.

OBJECTIVES: The aim of this study was to investigate how a moderate increase in dietary meat content combined (or not) with soluble fibre would influence protein digestion as well as digesta characteristics and flow. METHODS: Four groups of pigs were fed Western-style diets (high-protein/high-fat) containing two types of barbecued red meat, one with and one without a wheat arabinoxylan-rich fraction. After 4 wk, digesta samples were collected from small and large intestinal sites and analyzed for protein, amino acids, dry matter, and acid-insoluble ash. Tissue samples were also collected from each site. RESULTS: Arabinoxylan consumption led to somewhat lower apparent protein digestibility within the small and large intestines as well as shorter mean retention times. This suggests that the lowered protein digestibility is due, at least partly, to shorter access time to digestive proteases and absorptive surfaces. Additionally, digesta mass was higher in pigs fed arabinoxylan while dry matter (%) was lower, indicating an increased digesta water-holding capacity due to the presence of a soluble dietary fiber. CONCLUSION: Data showed that solubilized wheat arabinoxylan provides potential health benefits through decreased protein digestibility, increased digesta mass, and reduced mean retention time, even for diets with a moderately higher protein content. These factors are associated with efficiency of digestion and satiety, both of which have implications for prevention of obesity and other health disorders.

Butyrylated starch intake can prevent red meat-induced O6-methyl-2-deoxyguanosine adducts in human rectal tissue: a randomised clinical trial.

Epidemiological studies have identified increased colorectal cancer (CRC) risk with high red meat (HRM) intakes, whereas dietary fibre intake appears to be protective. In the present study, we examined whether a HRM diet increased rectal O(6)-methyl-2-deoxyguanosine (O(6)MeG) adduct levels in healthy human subjects, and whether butyrylated high-amylose maize starch (HAMSB) was protective. A group of twenty-three individuals consumed 300 g/d of cooked red meat without (HRM diet) or with 40 g/d of HAMSB (HRM+HAMSB diet) over 4-week periods separated by a 4-week washout in a randomised cross-over design. Stool and rectal biopsy samples were collected for biochemical, microbial and immunohistochemical analyses at baseline and at the end of each 4-week intervention period. The HRM diet increased rectal O(6)MeG adducts relative to its baseline by 21% (P < 0.01), whereas the addition of HAMSB to the HRM diet prevented this increase. Epithelial proliferation increased with both the HRM (P < 0.001) and HRM + HAMSB (P < 0.05) diets when compared with their respective baseline levels, but was lower following the HRM + HAMSB diet compared with the HRM diet (P < 0.05). Relative to its baseline, the HRM + HAMSB diet increased the excretion of SCFA by over 20% (P < 0.05) and increased the absolute abundances of the Clostridium coccoides group (P < 0.05), the Clostridium leptum group (P < 0.05), Lactobacillus spp. (P < 0.01), Parabacteroides distasonis (P < 0.001) and Ruminococcus bromii (P < 0.05), but lowered Ruminococcus torques (P < 0.05) and the proportions of Ruminococcus gnavus, Ruminococcus torques and Escherichia coli (P < 0.01). HRM consumption could increase the risk of CRC through increased formation of colorectal epithelial O(6)MeG adducts. HAMSB consumption prevented red meat-induced adduct formation, which may be associated with increased stool SCFA levels and/or changes in the microbiota composition.

A genetic strategy generating wheat with very high amylose content.

Resistant starch (RS), a type of dietary fibre, plays an important role in human health; however, the content of RS in most modern processed starchy foods is low. Cereal starch, when structurally manipulated through a modified starch biosynthetic pathway to greatly increase the amylose content, could be an important food source of RS. Transgenic studies have previously revealed the requirement of simultaneous down-regulation of two starch branching enzyme (SBE) II isoforms both located on the long arm of chromosome 2, namely SBEIIa and SBEIIb, to elevate the amylose content in wheat from ~25% to ~75%. The current study revealed close proximity of genes encoding SBEIIa and SBEIIb isoforms in wheat with a genetic distance of 0.5 cM on chromosome 2B. A series of deletion and single nucleotide polymorphism (SNP) loss of function alleles in SBEIIa, SBEIIb or both was isolated from two different wheat populations. A breeding strategy to combine deletions and SNPs generated wheat genotypes with altered expression levels of SBEIIa and SBEIIb, elevating the amylose content to an unprecedented ~85%, with a marked concomitant increase in RS content. Biochemical assays were used to confirm the complete absence in the grain of expression of SBEIIa from all three genomes in combination with the absence of SBEIIb from one of the genomes.

Wholegrain barley ß-glucan fermentation does not improve glucose tolerance in rats fed a high-fat diet.

Fermentation of oat and barley ß-glucans is believed to mediate in part their metabolic health benefits, but the exact mechanisms remain unclear. In this study, we sought to test the hypothesis that barley ß-glucan fermentation raises circulating incretin hormone levels and improves glucose control, independent of other grain components. Male Sprague-Dawley rats (n = 30) were fed a high-fat diet for 6 weeks and then randomly allocated to 1 of 3 dietary treatments for 2 weeks. The low- (LBG, 0% ß-glucan) and high- (HBG, 3% ß-glucan) ß-glucan diets contained 25% wholegrain barley and similar levels of insoluble dietary fiber, available carbohydrate, and energy. A low-fiber diet (basal) was included for comparison. Immediately prior to the dietary intervention, gastric emptying rate (using the (13)C-octanoic breath test) and postprandial glycemic response of each diet were determined. At the end of the study, circulating gut hormone levels were determined; and a glucose tolerance test was performed. The rats were then killed, and indices of cecal fermentation were assessed. Diet did not affect live weight; however, the HBG diet, compared to basal and LBG, reduced food intake, tended to slow gastric emptying, increased cecal digesta mass and individual and total short-chain fatty acid pools, and lowered digesta pH. In contrast, circulating levels of glucose, insulin, gastric-inhibitory peptide, and glucagon-like peptide-1, and glucose tolerance were unaffected by diet. In conclusion, wholegrain barley ß-glucan suppressed feed intake and increased cecal fermentation but did not improve postprandial glucose control or insulin sensitivity.