Brown seaweed as a food ingredient contributing to an adequate but not excessive amount of iodine in the European diet. A case study with bread.

BACKGROUND: The large amounts of iodine in brown seaweeds may contribute towards an adequate iodine intake, but also pose a food safety risk. In the current work we estimate the maximum amount of the cultivated brown seaweeds Saccharina latissima and Alaria esculenta (blanched and non-blanched) that can be added to white loaf-type wheat-bread so European consumers are protected against excessive chronic intakes. We use data for high-level consumers of bread with special ingredients added from the EFSA comprehensive European food consumption database to construct a conservative risk management model. We bake prototype seaweed bread and use the output from the model to assess exposure to iodine and trace metals. We also assess some bread quality parameters such as sensory characteristics, crumb firmness and specific volume. RESULTS: The maximum level of iodine in bread that would mitigate the European consumer from excess intakes was 857 µg I kg-1 bread. Assuming a typical 60% wheat flour per kilogram of bread, a maximum amount of 11.3 g of dried blanched A. esculenta (115 mg I kg-1 dry weight) could be incorporated per kilogram of wheat flour into the bread recipe whilst for non-blanched S. latissima, which contained 3500 mg I kg-1 dry weight, only a minuscule 350 mg of seaweed was the limit. For prototype breads, seaweed addition had no significant impact on the specific volume and crumb firmness. Levels of cadmium and arsenic in the bread were also of low toxicological concern. CONCLUSION: A maximum 1-2% brown seaweed of total flour weight could be added to bread to contribute towards European consumers' adequate iodine intake without overexposure. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A Three-Day Intervention With Granola Containing Cereal Beta-Glucan Improves Glycemic Response and Changes the Gut Microbiota in Healthy Individuals: A Crossover Study.

Intake of soluble fibers including beta-glucan, is known to improve post-prandial glycemic response. The mechanisms have been attributed to the viscous gel forming in the stomach and small intestine, giving a longer absorption time. However, recent evidence suggests a link between intake of beta-glucan and improved glycemic regulation at subsequent meals through the gut microbiota. We investigated the short-term effect of granola with different amounts of cereal beta-glucan on glycemic response and gut microbiota. After a two-week run-in period (baseline), fourteen healthy, normal weight adults completed a dose-response dietary crossover study. Different amounts of cereal beta-glucan (low: 0.8 g, medium: 3.2 g and high: 6.6 g) were provided in granola and eaten with 200 ml low-fat milk as an evening meal for three consecutive days. Blood glucose and insulin were measured fasted and after an oral glucose tolerance test (OGTT) the following day, in addition to peptide YY (PYY) and glucagon-like peptide (GLP-2), fasting short chain fatty acids (SCFA) in blood, breath H2, and gut microbiota in feces. Only the intervention with medium amounts of beta-glucan decreased blood glucose and insulin during OGTT compared to baseline. Fasting PYY increased with both medium and high beta-glucan meal compared to the low beta-glucan meal. The microbiota and SCFAs changed after all three interventions compared to baseline, where acetate and butyrate increased, while propionate was unchanged. Highest positive effect size after intake of beta-glucan was found with Haemophilus, followed by Veillonella and Sutterella. Furthermore, we found several correlations between different bacterial taxa and markers of glycemic response. In summary, intake of granola containing 3.2 g cereal beta-glucan as an evening meal for three consecutive days reduced the glycemic response after an OGTT 0-180 min and changed gut microbiota composition. Since we cannot rule out that other fiber types have contributed to the effect, more studies are needed to further explore the effect of cereal beta-glucan on glycemic regulation. Clinical Trial Registration: [www.clinicaltrials.gov], identifier [NCT03293693].

Improved estimation of in vitro protein digestibility of different foods using size exclusion chromatography.

While the harmonized INFOGEST model provides a physiologically relevant platform for simulated digestion, it needs to be combined with adequate analytical methods to enable quantification and comparison of protein digestibility in different food matrices. We have shown that size exclusion chromatography (SEC) can be used to estimate the proportion of small peptides potentially available for uptake. Combined with determination of total dissolved protein, the % of small peptides per total protein was calculated as a physiologically relevant estimate of protein digestibility (DSEC). Values for DSEC differed for casein (87.6%), chicken mince (72.6%), heated pea protein concentrate (67.8%), bread (63%), beef entrecote (57.7%) and pea protein concentrate (57.8%). In contrast to existing methods (TCA soluble protein, free NH2-groups), the proposed SEC based method gives separate insight into the two fundamental processes during protein digestion (solubilization and break-down), while maintaining the ability to rank digestibility of very different food proteins.

Advanced structural characterisation of agar-based hydrogels: Rheological and small angle scattering studies.

Agar-based extracts from Gelidium sesquipedale were generated by heat and combined heat-sonication, with and without the application of alkali pre-treatment. Pre-treatment yielded extracts with greater agar contents; however, it produced partial degradation of the agar, reducing its molecular weight. Sonication produced extracts with lower agar contents and decreased molecular weights. A gelation mechanism is proposed based on the rheological and small angle scattering characterization of the extracts. The formation of strong hydrogels upon cooling was caused by the association of agarose chains into double helices and bundles, the sizes of which depended on the agar purity and molecular weight. These different arrangements at the molecular scale consequently affected the mechanical performance of the obtained hydrogels. Heating of the hydrogels produced a gradual disruption of the bundles; weaker or smaller bundles were formed upon subsequent cooling, suggesting that the process was not completely reversible.

Peptides from chicken processing by-product inhibit DPP-IV and promote cellular glucose uptake: potential ingredients for T2D management.

Inhibition of dipeptidyl peptidase IV (DPP-IV) and stimulation of muscle glucose uptake are two of the key strategies for management of type-2-diabetes (T2D). In the present study, four protein hydrolysates generated by enzymatic hydrolysis of chicken by-product, i.e., mechanical chicken deboning residue, were evaluated for their DPP-IV inhibitory activity as well as their effect on glucose uptake by skeletal muscle cells. The DPP-IV inhibitory assay was performed at two concentrations (1000 µg mL-1 and 10 µg mL-1) for the crude chicken protein hydrolysates. The hydrolysate with the highest DPP-IV inhibition was selected for preparative-scale fractionation using size-exclusion chromatography (SEC). The SEC fractions were tested for DPP-IV inhibitory activity as well as their effect on glucose uptake and metabolic activity of skeletal muscle cells. The muscle cells were treated with the SEC fractions and glucose uptake was measured based on luminescence detection of 2-deoxyglucose-6-phosphate (2DG6P). A fraction with peptides in the lower molecular weight range was shown to promote glucose uptake and to inhibit DPP-IV. Further chromatographic fractionation followed by inhibition assaying of the most potent SEC fraction led to isolation of five refined peptide fractions with more than 80% DPP-IV inhibition, which were subsequently analyzed with LC-HRMS/MS. This led to identification of 14 peptides as potential DPP-IV inhibitors from protein hydrolysates of mechanical chicken deboning residue.

Predicting mixed-meal measured glycaemic index in healthy subjects.

PURPOSE: To determine the influence of meal composition on the glycaemic impact of different carbohydrate staples, and the accuracy of "adjusted calculated meal GI" compared with "measured mixed-meal GI". METHODS: In a non-blind randomized crossover trial fasted healthy subjects consumed four dinner-type mixed meals of realistic serving size comprising a carbohydrate staple of either mashed potato, pasta, rice or a glucose drink, combined with fixed portions of boiled carrots, poached salmon and herb sauce. Blood samples collected between 0 and 180 min were analysed for glucose and insulin concentrations. Adjusted calculated meal GI values were determined against a 50 g reference glucose drink, and compared to corresponding measured mixed-meal GIs, supplemented with data from four previous mixed-meal postprandial glycaemic response studies. RESULTS: The common carbohydrate staples, and the glucose drink, ingested as part of the salmon mixed meal induced a significantly lower post-prandial relative glycaemic response (RGR) and concurrent higher relative insulin response than the same amount of staple eaten alone. Adjusted calculated mixed-meal GI closely predicted measured mixed-meal GI in healthy subjects for 15 out of 17 mixed meals examined, showing the need to account for effects of fat and protein when predicting measured mixed-meal GI. Further, we showed the validity of using customarily consumed food amounts in mixed-meal postprandial RGR study design. CONCLUSIONS: Adjusted calculated mixed-meal GI appears a useful model to predict measured mixed-meal GI in healthy subjects and with further development and validation could aid nutrition research and rational design of healthy meals for personalized nutrition and particular consumer groups.

Effects of pH on steam explosion extraction of acetylated galactoglucomannan from Norway spruce.

BACKGROUND: Acetylated galactoglucomannan (AcGGM) is a complex hemicellulose found in softwoods such as Norway spruce (Picea abies). AcGGM has a large potential as a biorefinery feedstock and source of oligosaccharides for high-value industrial applications. Steam explosion is an effective method for extraction of carbohydrates from plant biomass. Increasing the reaction pH reduces the combined severity ( R 0 ' ) of treatment, affecting yields and properties of extracted oligosaccharides. In this study, steam explosion was used to extract oligosaccharides from Norway spruce wood chips soaked with sodium citrate and potassium phosphate buffers with pH of 4.0-7.0. Yields, monosaccharide composition of released oligosaccharides and biomass residue, their acetate content and composition of their lignin fraction were examined to determine the impact of steam explosion buffering on the extraction of softwood hemicellulose. RESULTS: Reducing the severity of steam explosion resulted in lower yields, although the extracted oligosaccharides had a higher degree of polymerization. Higher buffering pH also resulted in a higher fraction of xylan in the extracted oligos. Oligosaccharides extracted in buffers of pH > 5.0 were deacetylated. Buffering leads to a removal of acetylations from both the extracted oligosaccharides and the hemicellulose in the residual biomass. Treatment of the residual biomass with a GH5 family mannanase from Aspergillus nidulans was not able to improve the AcGGM yields. No hydroxymethylfurfural formation, a decomposition product from hexoses, was observed in samples soaked with buffers at pH higher than 4.0. CONCLUSIONS: Buffering the steam explosion reactions proved to be an effective way to reduce the combined severity ( R 0 ' ) and produce a wide range of products from the same feedstock at the same physical conditions. The results highlight the impact of chemical autohydrolysis of hemicellulose by acetic acid released from the biomass in hydrothermal pretreatments. Lower combined severity results in products with a lower degree of acetylation of both the extracted oligosaccharides and residual biomass. Decrease in severity appears not to be the result of reduced acetate release, but rather a result of inhibited autohydrolysis by the released acetate. Based on the results presented, the optimal soaking pH for fine-tuning properties of extracted AcGGM is below 5.0.

Glyceamic and insulinaemic response to mashed potato alone, or with broccoli, broccoli fibre or cellulose in healthy adults.

PURPOSE: To examine the role of realistic serving sizes of broccoli, broccoli fibre and cellulose co-consumed with mash potato, or mashed potato eaten alone, on glycaemic and insulinaemic responses (GR and IR) in healthy adults. METHOD: A non-blind randomized crossover trial was conducted with thirteen healthy subjects consuming four different meals. Capillary blood samples between 0 and 180 min were analysed for glucose and insulin. The incremental area under the fasting blood glucose and insulin curves (iAUC) was calculated for different time increments. Differences in GR and IR between meals were assessed by repeated measures analysis of variance. RESULTS: The immediate GR and IR to one serving of mashed potato eaten with two servings of broccoli were significantly lower than mashed potato eaten alone. The peak, incremental peak and iAUC0-30min for GR and iAUC0-30min for IR were all significantly lower for the broccoli-potato meal. This meal also takes longer to return to fasting baseline with a time-delayed lag in IR and GR compared to the potato only meal. The iAUC60-120min for IR was significantly greater for the broccoli-potato meal compared to the other meals. Yet there was no corresponding significant difference between the broccoli-potato meal and the other meals for peak, incremental peak IR or any other iAUCs for GR and IR. For the potato meals containing added broccoli fibre or cellulose, no significant differences in GR or IR were observed when compared with the potato eaten alone. CONCLUSION: Co-consumption of cooked broccoli with mashed potato has a significant effect on glycaemic and insulinaemic responses compared to potato eaten alone. Our study suggests broccoli eaten with potato improves glucose homeostasis and therefore indicates a general beneficial nutritional role for broccoli when eaten with a carbohydrate staple.

Effect of Dietary Fibers on Cecal Microbiota and Intestinal Tumorigenesis in Azoxymethane Treated A/J Min/+ Mice.

Foods naturally high in dietary fiber are generally considered to protect against development of colorectal cancer (CRC). However, the intrinsic effect of dietary fiber on intestinal carcinogenesis is unclear. We used azoxymethane (AOM) treated A/J Min/+ mice, which developed a significantly higher tumor load in the colon than in the small intestine, to compare the effects of dietary inulin (IN), cellulose (CE) or brewers spent grain (BSG) on intestinal tumorigenesis and cecal microbiota. Each fiber was tested at two dose levels, 5% and 15% (w/w) content of the AIN-93M diet. The microbiota was investigated by next-generation sequencing of the 16S rRNA gene (V4). We found that mice fed IN had approximately 50% lower colonic tumor load than mice fed CE or BSG (p<0.001). Surprisingly, all three types of fiber caused a dose dependent increase of colonic tumor load (p<0.001). The small intestinal tumor load was not affected by the dietary fiber interventions. Mice fed IN had a lower bacterial diversity than mice fed CE or BSG. The Bacteroidetes/Firmicutes ratio was significantly (p = 0.003) different between the three fiber diets with a higher mean value in IN fed mice compared with BSG and CE. We also found a relation between microbiota and the colonic tumor load, where many of the operational taxonomic units (OTUs) related to low tumor load were significantly enriched in mice fed IN. Among the OTUs related to low tumor load were bacteria affiliated with the Bacteroides genus. These results suggest that type of dietary fiber may play a role in the development of CRC, and that the suppressive effect of IN on colonic tumorigenesis is associated with profound changes in the cecal microbiota profile.

Inter-laboratory evaluation of SEC-post-column calcofluor for determination of the weight-average molar mass of cereal ß-glucan.

Even though size exclusion chromatography (SEC) with post column addition of calcofluor (SEC-calcofluor) has been used for the determination of cereal ß-glucan molar mass in foods for many years, there is a lack of systematic evaluation of the method. To address this issue a set of suitable ß-glucan standards were generated by preparative SEC and their molar mass characteristics were determined by analytical multi-detection SEC (refractive index (RI), light scattering). Each standard was then analysed by SEC-calcofluor at three different labs. As a direct comparison, the analyses were repeated with a RI detector. For SEC-calcofluor accurate measurements of weight average molar mass (Mw) can be made for ß-glucan populations within 10-500×10(3)g/mol. Above this molar mass threshold there is an increasing tendency for underestimation of Mw. Precipitation of some ß-glucan-calcofluor complexes may have delayed their transport into the detector.

Viscosity based quantification of endogenous ß-glucanase activity in flour.

High molecular weight (MW) is a key parameter for cereal ß-glucans physiological benefits like decreased serum cholesterol and attenuated post prandial blood glucose. However, the activity of endogenous flour enzymes during bread production results in a decrease of ß-glucan MW. The depolymerization of a standard ß-glucan solution by different flour extracts (wheat, barley and rye) was followed by measuring the viscosity decrease with a Rheometer. The slope of the inverse viscosity (1/η) against degradation time was used to quantify ß-glucanase activity by comparison with slopes obtained with known concentrations of the ß-glucanase Lichenase. Results correlated well with depolymerization rates estimated by HPSEC. The viscosity based method is rapid (20 min per sample), accurate (≤6% variation), and a powerful screening tool for identifying flour fractions with low ß-glucanase activity, treatments that can inactivate ß-glucanases in flour, or the development of ß-glucanase inhibitors for the use in e.g. bread making.

Generic tools to assess genuine carbohydrate specific effects on in vitro immune modulation exemplified by ß-glucans.

Even if carbohydrate preparations from plant/fungal sources have a high degree of purity, observed immune-stimulation may be caused by minute sample contaminations. Using the example of different ß-glucans we present a range of analytical tools crucial for validation of possible immune-stimulatory effects. Two yeast (MacroGard and Zymosan) and one cereal ß-glucan (CBG40) increased IL-8 secretion by HT-29 cells considerably. Degradation of the ß-glucan samples with ß-glucan specific enzymes did hardly influence the effect of Zymosan and CBG40 but significantly decreased the effect of MacroGard. Stimulation of IL-8 secretion by CBG40 and Zymosan was hence not due to their ß-glucan content. Instead, the effect of the CBG40 sample was due to low levels of LPS despite the inability of the known LPS inhibitor Polymyxin B to supress its stimulatory effect. We conclude that targeted enzymatic degradation of samples is a powerful validation tool to investigate carbohydrate specific immune-modulation.

Application of high-performance anion-exchange chromatography with pulsed amperometric detection to compare the kinetic properties of ß-glucosidase on oligosaccharides from lichenase digested barley ß-glucan.

A chromatographic method using HPAEC-PAD was developed to accurately quantify the major oligosaccharides derived from lichenase degradation of barley ß-glucan. This method was further used to follow ß-glucosidase degradation and product formation as progress curves. This approach allowed us to compare the kinetic characteristics of ß-glucosidase on each exclusive oligosaccharide substrate and their mixtures. Our results show that when determining the kinetic parameters of exohydrolases on oligosaccharides following the progress curve for the substrates is necessary since calculations based only on released monosaccharide products may lead to an error. The catalytic activity of almond ß-glucosidase on laminaribiose (G3G) was approximately half that measured against cellobiose (3.15 S(-1)). The enzyme had 12 times and 560 times less catalytic activity on 3-O-ß-cellobiosyl-D-glucose (G4G3G) and on 3-O-ß-cellotriosyl-d-glucose (G4G4G3G) respectively than on G3G. Our approach offers a useful tool for the determination of the kinetics of enzymatic or chemical modification of various carbohydrate substrates.

Release and characterization of single side chains of white cabbage pectin and their complement-fixing activity.

A mixture of single side chains from white cabbage pectin were obtained by anion exchange chromatography after applying mild chemical conditions promoting beta-elimination. These pectin fragments were characterized by their molecular weight distribution, sugar composition, 13C-NMR, and MALDI-TOF-MS analysis. These analyses revealed that the large oligosaccharides released by beta-eliminative treatment were composed of alpha-1,5 linked arabinosyl residues with 2- and 3-linked alpha-arabinosyl side chains, and, or beta-1,4 linked galactosyl side chains. Fractions were tested for complement-fixing activity in order to determine their interaction with the complement system. These results strongly indicated that there was a minimal unit size responsible for the complement-fixing activity. Neutral pectin fragments (8 kDa) obtained from beta-elimination were inactive in the complement system, although they contained a sugar composition previously shown to be highly active. Larger pectin fragments (17 kDa) retained some activity, but much lower than polymers containing rhamnogalacturonan type 1 (RGI) structures isolated from the same source. This implied that structural elements containing multiple side chains is necessary for efficient complement-fixing activity.

Screening of acrylamide contents in potato crisps using process variable settings and near-infrared spectroscopy.

In order to study the formation of acrylamide in potato crisps during processing, an experimental design was set up. The design variables were drying time (6 levels), frying temperature (2 levels) and frying time (8 levels). The design contained 36 samples, which were analysed for acrylamide contents using LC high-resolution mass spectroscopy (LC-HRMS), and fat contents using the Soxhlet apparatus. Prior to analysis, all potato crisp samples were ground and analysed on an NIRSystems 6500 near-infrared (NIR) spectrometer. The acrylamide contents were modelled by: (i) design variables using multiple linear regression, (ii) NIR spectra using partial least squares regression (PLSR) and (iii) design variables and NIR spectra in combination using a novel technique combining least squares regression on the former, and PLSR on the latter. The results showed that the NIR spectra alone or in combination with the design variables gave better prediction models for acrylamide than the design variables alone. This implies that the spectra contain chemical information that is not purely a result of the processing variables that were investigated in this experiment. NIR spectroscopy is proposed as a possible tool for screening and identification of potato crisps with a high acrylamide content.

Treatment of cereal products with a tailored preparation of trichoderma enzymes increases the amount of soluble dietary fiber.

Nutritionists recommend increasing the intake of soluble dietary fiber (SDF), which is very low in most cereal-based products. Conversion of insoluble DF (IDF) into SDF can be achieved by chemical treatments, but this affects the sensorial properties of the products. In this study, the possibility of getting a substantial increase of SDF from cereal products using a tailored preparation of Trichoderma enzymes is reported. Enzymes were produced cultivating Trichoderma using durum wheat fiber (DWF) and barley spent grain (BSG) as unique carbon sources. Many Trichoderma strains were screened, and the hydrolysis conditions able to increase by enzymatic treatment the amount of SDF in DWF and BSG were determined. Results demonstrate in both products that it is possible to triple the amount of SDF without a marked decrease of total DF. The enzymatic treatment also causes the release of hydroxycinnamic acids, mainly ferulic acid, that are linked to the polysaccharides chains. This increases the free phenolic concentration, the water-soluble antioxidant activity, and, in turn, the phenol compounds bioavailability.

Pectin isolated from white cabbage--structure and complement-fixing activity.

This study was done to investigate whether white cabbage contained polysaccharides with immunostimulatory activity using the complement-fixing test as an indicator. The main polysaccharide isolated was of pectin nature. Methanolysis and (13)C-NMR showed that the polymers consisted of highly esterified alpha-galactopyranoside (alpha-GalpA), significant amounts of alpha-arabinose furanoside (alpha-Araf), beta-Galp and lesser amounts of rhamnose in the pyranose form (Rhap) and xylose in the pyranose form (Xylp). Linkage analyses showed that the alpha-GalpA residues were mainly 1,4-linked with small amounts of 1,3,4-linkages. The alpha-Araf residues were mainly terminally (t)- and 1,5-linked, whereas beta-Galp was t-, 1,3-, 1,6-, and 1,3,6-linked. Positive Yariv reaction indicated polymers with arabinogalactan type 2 like structures. alpha-Rhap was mainly present as 1,2- and 1,2,4-linked residues and Xylp was t- and 1,4-linked. The molecular weight varied greatly and was from 10 to 150 kDa. Cabbage polymers had biological activity and this complement-fixing activity was greatly affected by hydrolytic removal of Araf from pectic side chains.

Flavanol and bound phenolic acid contents in different barley varieties.

Different barley varieties, consisting of hulled and hull-less types, of normal, waxy, and high amylose starch, as well as two-rowed and six-rowed types, were analyzed for their main proanthocyanidins and bound phenolic acids. Variations in proanthocyanidin and phenolic acid contents were studied in different barley types as well as inter-relationships between the phytochemicals and polysaccharides. The main flavanols found in the analyzed barley varieties were two dimeric as well as four trimeric forms in addition to catechin. The total amount of flavanols ranged from 325 to 527 microg/g of fresh weight of barley flour. No evident associations were found between variations in proanthocyanidin levels and different barley types. The total amount of phenolic acids ranged from 604 to 1346 microg/g of fresh weight of barley flour, with ferulic acid as the dominating acid. The amount of phenolic acids varied according to occurrence or lack of hull, with significantly higher levels in the hulled varieties.

Addition of glycine reduces the content of acrylamide in cereal and potato products.

The effects of adding amino acids on the content of acrylamide in potato crisps, French fries, flat breads, and bread crusts were investigated. Addition of glycine or glutamine during blanching of crisps reduced the amount of acrylamide by approximately 30% compared to no addition. No effect was found in French fries. Addition of glycine during doughmaking significantly reduced acrylamide in both flat breads and bread crusts. In bread crusts the reduction of acrylamide ranged from 50 to >90% depending on the baking condition. In flat breads the reduction varied between 60 and >95%.

Effective ways of decreasing acrylamide content in potato crisps during processing.

The aim of this work was to examine the effect of blanching or soaking in different acid solutions on the acrylamide content in potato crisps. Furthermore, the effects of a shorter frying time and a lower frying temperature combined with a postdrying were investigated. Soaking or blanching of potato slices in acidic solutions decreased the pH of potato juice and increased the extraction of amino acids and sugars. Potato crisps obtained after such pretreatments were characterized by lower acrylamide content. The most effective extraction of free amino acids and sugars as well as the largest decrease of acrylamide content (90%) in crisps was obtained when potato slices were soaked in acetic acid solution for 60 min at 20 degrees C. Shorter frying time followed by postdrying resulted in low-moisture potato crisps. Furthermore, the postdrying treatment gave a decreases in acrylamide content of approximately 70% when potato slices were fried at 185 degrees C and approximately 80% when potato slices were fried at 160 degrees C. Effective ways of decreasing acrylamide content in crisps production have been found. Crisps with low acrylamide content and good sensory quality can be obtained either by blanching in acetic acid as pretreatment or by a short frying followed by postdrying.