Dr Terri Grassby

Faba bean (Vicia faba) is an underutilised leguminous crop that is rich in protein and carbohydrates with the potential to improve food and nutrition security globally. The beans are equally a good source of fibre, antioxidants, minerals and vitamins. Besides the nutritional value of the grains, Faba bean also contributes to maintaining the sustainability of agricultural systems through crop rotation for soil enrichment and in the symbiotic fixation of atmospheric nitrogen. The bulk of carbohydrate in the crop is starch, which may vary between approximately 33 and 48% depending on variety and extraction method. With the growing demand for new starch sources, Faba bean starch could serve as an alternative to conventional corn and potato starches. This chapter presents the physicochemical, thermal, structural, and in vitro digestibility properties of Faba bean starch. It also discusses the different modification methods used to enhance the thermal and physicochemical properties of the starch isolate, as well as the potential industrial applications. Future studies on the native and modified Faba bean starches are also suggested.

Dietary fibre is a generic term for a chemically diverse group of carbohydrates that are resistant to endogenous enzymes of the human digestive tract. The major component of dietary fibre consists of plant cell walls, which are supramolecular structures, composed of complex heterogeneous networks of cellulose, hemicelluloses and pectic substances. The amounts and relative proportions of these carbohydrates vary depending on the type and maturity of the plant tissue. Some plant cell walls, especially those from leguminous seeds, are rich in water-soluble non-starch polysaccharides (NSP). Intact plant cell walls and NSP affect the rate and extent of nutrient digestion, with important implications for health and disease. Certain types of fibre reduce the rate of starch digestion, which in turn can significantly attenuate the postprandial rise in blood glucose and insulin concentrations. This is potentially beneficial in the prevention and treatment of diseases, including diabetes mellitus and cardiovascular disease. However, the mechanisms of action of NSP in relation to the digestive process are still not well understood. They are thought to include formation of viscous solutions, encapsulation of nutrients and inhibition of digestive enzymes. These mechanisms are illustrated using specific examples. Oat β-glucan is used to show the effects of various processing techniques on β-glucan molecular weight, and hence viscosity, on risk factors for diabetes and cardiovascular disease. Evidence for guar galactomannan acting as an inhibitor of α-amylase, in addition to forming viscous solutions, is presented. Finally, the effect of intact plant cell walls on the bioaccessibility of nutrients is discussed.

Chinese water chestnut (Eleocharis dulcis (Burman f.) Trin ex Henschel) is a corm consumed globally in Oriental-style cuisine. The corm consists of three main tissues, the epidermis, subepidermis, and parenchyma; the cell walls of which were analyzed for sugar, phenolic, and lignin content. Sugar content, measured by gas chromatography, was higher in the parenchyma cell walls (931 μg/mg) than in the subepidermis (775 μg/mg) or epidermis (685 μg/mg). The alkali-extractable phenolic content, measured by high-performance liquid chromatography, was greater in the epidermal (32.4 μg/mg) and subepidermal cell walls (21.7 μg/mg) than in the cell walls of the parenchyma (12.3 μg/mg). The proportion of diferulic acids was higher in the parenchyma. The Klason lignin content of epidermal and subepidermal cell walls was ~15%. Methylation analysis of Chinese water chestnut cell-wall polysaccharides identified xyloglucan as the predominant hemicellulose in the parenchyma for the first time, and also a significant pectin component, similar to other nongraminaceous monocots.

Investigators often study product release from starches during prolonged incubations with α-amylase in vitro. The reaction time courses usually fit to a linear form of a first order rate equation, i.e., ln[(C∞ − Ct)/C∞] = −kt. This equation calls for an accurate estimate of C∞, i.e., the concentration of product at the end of the reaction. Estimates of C∞ from digestibility curves can be unreliable. The Guggenheim method does not require prior knowledge of C∞ but seems not to have been applied to starch hydrolysis data. An alternative method is also available in which the logarithm of the slope (LOS) of a digestibility curve at various time points is plotted against time. This allows estimations of both k and C∞ and can also reveal whether changes occur in digestion rate from rapid to slow as digestion proceeds. We describe the Guggenheim and LOS methods and provide examples of their application to starch digestibility data.

A number of studies have demonstrated that consuming almonds increases satiety but does not result in weight gain, despite their high energy and lipid content. To understand the mechanism of almond digestion, in the present study, we investigated the bioaccessibility of lipids from masticated almonds during in vitro simulated human digestion, and determined the associated changes in cell-wall composition and cellular microstructure. The influence of processing on lipid release was assessed by using natural raw almonds (NA) and roasted almonds (RA). Masticated samples from four healthy adults (two females, two males) were exposed to a dynamic gastric model of digestion followed by simulated duodenal digestion. Between 7·8 and 11·1 % of the total lipid was released as a result of mastication, with no significant differences between the NA and RA samples. Significant digestion occurred during the in vitro gastric phase (16·4 and 15·9 %) and the in vitro duodenal phase (32·2 and 32·7 %) for the NA and RA samples, respectively. Roasting produced a smaller average particle size distribution post-mastication; however, this was not significant in terms of lipid release. Light microscopy showed major changes that occurred in the distribution of lipid in all cells after the roasting process. Further changes were observed in the surface cells of almond fragments and in fractured cells after exposure to the duodenal environment. Almond cell walls prevented lipid release from intact cells, providing a mechanism for incomplete nutrient absorption in the gut. The composition of almond cell walls was not affected by processing or simulated digestion.

Although the growth of bacteria has been studied for more than a century, it is only in recent decades that surface-associated growth has received attention. In addition to the well-characterized biofilm and swarming lifestyles, bacteria can also develop as micro-colonies supported by structured environments in both food products and the GI tract. This immobilized mode of growth has not been widely studied. To develop our understanding of the effects of immobilization upon a food-borne bacterial pathogen, we used the IFR Gel Cassette model. The transcriptional programme and metabolomic profile of Salmonella enterica serovar Typhimurium ST4/74 were compared during planktonic and immobilized growth, and a number of immobilization-specific characteristics were identified. Immobilized S.Typhimurium did not express motility and chemotaxis genes, and electron microscopy revealed the absence of flagella. The expression of RpoS-dependent genes and the level of RpoS protein were increased in immobilized bacteria, compared with planktonic growth. Immobilized growth prevented the induction of SPI1, SPI4 and SPI5 gene expression, likely mediated by the FliZ transcriptional regulator. Using an epithelial cell-based assay, we showed that immobilized S.Typhimurium was significantly less invasive than planktonic bacteria, and we suggest that S.Typhimurium grown in immobilized environments are less virulent than planktonic bacteria. Our findings identify immobilization as a third type of surface-associated growth that is distinct from the biofilm and swarming lifestyles of Salmonella.

food matrix (muffins) investigating whether the cell-wall barrier regulates the bioaccessibility of nutrients within this matrix. Muffins containing small (AF) or large (AP) particles of almond were digested in triplicate using an in vitro dynamic gastric model (DGM, 1 h) followed by a static duodenal digestion (8 h). AF muffins had 97.1 ± 1.7% of their lipid digested, whereas AP muffins had 57.6 ± 1.1% digested. In vivo digestion of these muffins by an ileostomy volunteer (0–10 h) gave similar results with 96.5% and 56.5% lipid digested, respectively. The AF muffins produced a higher postprandial triacylglycerol iAUC response (by 61%) than the AP muffins. Microstructural analysis showed that some lipid remained encapsulated within the plant tissue throughout digestion. The cell-wall barrier mechanism is the main factor in regulating lipid bioaccessibility from almond particles.

We have previously reported on the low lipid bioaccessibility from almond seeds during digestion in the upper gastrointestinal tract (GIT). In the present study, we quantified the lipid released during artificial mastication from four almond meals: natural raw almonds (NA), roasted almonds (RA), roasted diced almonds (DA) and almond butter from roasted almonds (AB). Lipid release after mastication (8.9% from NA, 11.8% from RA, 12.4% from DA and 6.2% from AB) was used to validate our theoretical mathematical model of lipid bioaccessibility. The total lipid potentially available for digestion in AB was 94.0%, which included the freely available lipid resulting from the initial sample processing and the further small amount of lipid released from the intact almond particles during mastication. Particle size distributions measured after mastication in NA, RA and DA showed most of the particles had a size of 1000 µm and above, whereas AB bolus mainly contained small particles (

BACKGROUND: Cereal crops, particularly wheat, are a major dietary source of starch, and the bioaccessibility of starch has implications for postprandial glycemia. The structure and properties of plant foods have been identified as critical factors in influencing nutrient bioaccessibility; however, the physical and biochemical disassembly of cereal food during digestion has not been widely studied. OBJECTIVES: The aims of this study were to compare the effects of 2 porridge meals prepared from wheat endosperm with different degrees of starch bioaccessibility on postprandial metabolism (e.g., glycemia) and to gain insight into the structural and biochemical breakdown of the test meals during gastroileal transit. DESIGN: A randomized crossover trial in 9 healthy ileostomy participants was designed to compare the effects of 55 g starch, provided as coarse (2-mm particles) or smooth (

BACKGROUND: The particle size and structure of masticated almonds have a significant impact on nutrient release (bioaccessibility) and digestion kinetics. OBJECTIVES: The goals of this study were to quantify the effects of mastication on the bioaccessibility of intracellular lipid of almond tissue and examine microstructural characteristics of masticated almonds. DESIGN: In a randomized, subject-blind, crossover trial, 17 healthy subjects chewed natural almonds (NAs) or roasted almonds (RAs) in 4 separate mastication sessions. Particle size distributions (PSDs) of the expectorated boluses were measured by using mechanical sieving and laser diffraction (primary outcome). The microstructure of masticated almonds, including the structural integrity of the cell walls (i.e., dietary fiber), was examined with microscopy. Lipid bioaccessibility was predicted by using a theoretical model, based on almond particle size and cell dimensions, and then compared with empirically derived release data. RESULTS: Intersubject variations (n = 15; 2 subjects withdrew) in PSDs of both NA and RA samples were small (e.g., laser diffraction; CV: 12% and 9%, respectively). Significant differences in PSDs were found between these 2 almond forms (P < 0.05). A small proportion of lipid was released from ruptured cells on fractured surfaces of masticated particles, as predicted by using the mathematical model (8.5% and 11.3% for NAs and RAs, respectively). This low percentage of lipid bioaccessibility is attributable to the high proportion (35-40%) of large particles (>500 μm) in masticated almonds. Microstructural examination of the almonds indicated that most intracellular lipid remained undisturbed in intact cells after mastication. No adverse events were recorded. CONCLUSIONS: Following mastication, most of the almond cells remained intact with lipid encapsulated by cell walls. Thus, most of the lipid in masticated almonds is not immediately bioaccessible and remains unavailable for early stages of digestion. The lipid encapsulation mechanism provides a convincing explanation for why almonds have a low metabolizable energy content and an attenuated impact on postprandial lipemia.

Polysaccharides derived from plant foods are major components of the human diet, with limited contributions of related components from fungal and algal sources. In particular, starch and other storage carbohydrates are the major sources of energy in all diets, while cell wall polysaccharides are the major components of dietary fibre. We review the role of these components in the human diet, including their structure and distribution, their modification during food processing and effects on functional properties, their behaviour in the gastro-intestinal tract and their contribution to healthy diets.

Molecular simulations can measure strain in bond lengths, angles, torsion, and non-bonded interactions. High strain energy indicates a reactive and unstable molecule. Method 4-turn amylose and saturated FA models (C8-C22) were built using MOE2020.09 software. Molecular dynamics were simulated in water, with complexes containing 1 or 2 FAs per amylose helix. Results Conclusion The lower the energy, the more stable the molecule. • Double FA complexes are less stable than single FA complexes. • Trend: The stability increases from C8 and decreases from C14. • C10 single FA complexes have the lowest energy so are most likely to be stable, meaning they are a feasible candidate for practical use.

Amylose, the linear fraction of starch, can form single helix inclusion complexes with a range of guest molecules including lipids, iodine, alcohols and flavour compounds(Reference Li, Liu and Zhang1). Amylose-lipid complexes (ALCs) are known as resistant starch 5. When eaten in food such as bread, potato or pasta, resistant starches withstand digestion by ensymes in the small intestine and pass into the colon where they act as substrates for fermentation by the gut microbiota(Reference Annison and Topping2). Starch digestion in the small intestine is critical in determining post-prandial blood glucose and insulin concentrations, with wider health implications in development and management of metabolic and cardiovascular diseases(Reference Hasjim, Lee and Hendrich3). This includes lower blood glucose and cholesterol levels, lower risk of colo-rectal cancers and cardiometabolic diseases, and increased satiety(Reference Panyoo and Emmambux4). Enhancement of starchy foods, for example pasta, with ALCs would bring a host of health benefits, and molecular modelling can be used to gain greater understanding of the structure and behaviour of ALCs in order to optimise their incorporation into food matrices. The aim of this study was to use molecular modelling to identify whether ALCs were more stable with one or two fatty acids, and which fatty acid gave the most stable complex. Molecular Mechanics was performed using the AMBER forcefield to build ALC molecules and measure their strain energy under static conditions. A 24 glucose-residue V-amylose helix was modelled with saturated fatty acids (even numbers C8-C22), and the complex binding energy calculated to determine the optimum amylose-fatty acid configuration. Molecular Dynamics simulations were run on the same molecules solvated in a 55Å water cube, then the binding energy calculated. When there is one saturated fatty acid in the complex, potential energy decreases as the hydrocarbon tail length increases, showing an increase in stability. However, when two fatty acids are in the complex, stability increases up to C16 (palmitic acid) then decreases for the longer chains. Once solvated, single fatty acids reach an optimum at C12 (lauric acid) and then stability decreases. Double fatty acids increase in stability up to C20 (arachidic acid), then decrease at C22. Electrostatic interactions also peaked at C20 and dropped at C22, indicating that they have a stabilising effect on the complexes. These results will inform which ALCs are produced in the lab to stand the greatest likelihood of being stable in foods, so they do not breakdown before reaching the small intestine of consumers. This optimum was C20 for ALCs containing two fatty acids. Further studies will look at the impact of the length of V-amylose polymers on formation of ALCs and on mixed combinations of fatty acids.

Almonds are rich in unsaturated lipids, which play a role in some of the reported benefits of almond consumption for human health. Almond lipids are poorly bioaccessible due to almonds’ unique physicochemical properties that influence particle size distribution (PSD) following mastication, allowing much intracellular lipid to escape digestion in the upper gastrointestinal tract. To investigate the impact of commercial processing (grinding almonds into flour), on PSD and predicted lipid bioaccessibility following mastication, a randomised cross-over design mastication study was conducted in healthy adults. The PSDs of masticated whole and ground almonds was assessed using two laboratory methods (mechanical sieving and laser diffraction). PSD from mechanical sieving was used to calculate lipid bioaccessibility using a theoretical mathematical model. Thirty-one healthy adults (18–45 years) completed both mastication sessions. Following mastication, ground almonds had a PSD with significantly fewer larger particles and more smaller particles, compared with whole almonds. Predicted lipid bioaccessibility of masticated ground almonds (10.4%, SD 1.8) was marginally but significantly greater than the predicted lipid bioaccessibility of masticated whole almonds (9.3%, SD 2.0; p = 0.017). Commercial grinding of almonds significantly influences the PSD of almonds following mastication, which results in a modest but significant increase in predicted lipid bioaccessibility. (This article belongs to the Special Issue Nut, Nut Products and Health)

Processed foods are increasingly under the spotlight since the development of classification systems based on proxies for food processing. Published critical reviews and commentaries suggest different views among professional disciplines about the definition and classification of processed food. There is a need to further understand perspectives of professionals on the conceptualisation of processed food and the agreements and disagreements among experts, to encourage interdisciplinary dialogue and aid communication to the public. The aim of this research was to elicit views and understandings of professionals on processed food, their perceptions of lay people's perceptions of the same, and their perspectives on the challenges of communicating about processed foods to the public. The online discussion groups brought together a range of professionals (n = 27), covering the fields of nutrition, food technology, policy making, industry, and civil society, mixed in 5 heterogenous groups. Through thematic analysis the following themes relating to the conceptualisation of processed food and challenges for communication were identified: (1) Broad concepts that need differentiation; (2) Disagreements on scope and degree of processing; (3) The role of food processing within the food system: the challenges in framing risks and benefits; and (4) The challenge of different perspectives and interests for risk communication. Throughout the discussions blurred lines in the characterisation of processing, processed foods, and unhealthy foods were observed. Participants agreed that consensus is important, but difficult. Participants identified a need for further interdisciplinary dialogue, including public engagement, to break down the observed issues, and work towards a mutual understanding and develop clear communication messages.

BACKGROUND Almonds contain lipid, fiber and polyphenols and possess physicochemical properties that impact nutrient bioaccessibility, which are hypothesized to impact gut physiology and microbiota. OBJECTIVES Investigate the impact of whole almonds and ground almonds (almond flour) on fecal bifidobacteria (primary outcome), gut microbiota composition and transit time. DESIGN Healthy adults (n = 87) participated in a parallel, 3-arm randomized controlled trial. Participants received whole almonds (56 g/d), ground almonds (56 g/d) or an isocaloric control muffin in place of habitual snacks for 4 weeks. Gut microbiota composition and diversity (16S rRNA gene sequencing), short-chain fatty acids (gas-chromatography), volatile organic compounds (gas-chromatography mass-spectrometry), gut transit time (wireless motility capsule), stool output and gut symptoms (7-day diary) were measured at baseline and endpoint. The impact of almond form on particle size distribution (PSD) and predicted lipid release was measured in a subgroup (n = 31). RESULTS Modified intention-to-treat analysis was performed on 79 participants. There were no significant differences in abundance of fecal bifidobacteria following consumption of whole almonds (8.7%, SD 7.7%), ground almonds (7.8%, SD 6.9%) or control (13.0%, SD 10.2%; q = 0.613). Consumption of almonds (whole and ground pooled) resulted in higher butyrate (24.1 μmol/g, SD 15.0 μmol/g) in comparison to control (18.2 μmol/g, SD 9.1 μmol/g; p = 0.046). There was no effect of almonds on gut microbiota at the phylum level or diversity, gut transit time, stool consistency or gut symptoms. Almond form (whole versus ground) had no effect on study outcomes. Ground almonds resulted in significantly smaller PSD and higher predicted lipid release (10.4%, SD 1.8%) in comparison to whole almonds (9.3%, SD 2.0%; p = 0.017). CONCLUSIONS Almond consumption has limited impact on gut microbiota composition but increases butyrate concentrations in adults, suggesting positive alterations to microbiota functionality. Almonds can be incorporated into the diet to increase fiber consumption without triggering gut symptoms. Clinical trial registry: ClinicalTrials.gov identifier - NCT03581812.

Background: Processed foods are typically praised/revered for their convenience, palatability, and novelty; however, their healthfulness has increasingly come under scrutiny. Classification systems that categorise foods according to their “level of processing” have been used to predict diet quality and health outcomes and inform dietary guidelines and product development. However, the classification criteria used are ambiguous, inconsistent and often give less weight to existing scientific evidence on nutrition and food processing effects; critical analysis of these criteria creates conflict amongst researchers. Scope and approach: We examine the underlying basis of food classification systems and provide a critical analysis of their purpose, scientific basis, and distinguishing features by thematic analysis of the category definitions. Key findings and conclusions: These classification systems were mostly created to study the relationship between industrial products and health. There is no consensus on what factors determine the level of food processing. We identified four defining themes underlying the classification systems: 1. Extent of change (from natural state); 2. Nature of change (properties, adding ingredients); 3. Place of processing (where/by whom); and 4. Purpose of processing (why, essential/cosmetic). The classification systems embody socio-cultural elements and subjective terms, including home cooking and naturalness. Hence, “processing” is a chaotic conception, not only concerned with technical processes. Most classification systems do not include quantitative measures but, instead, imply correlation between “processing” and nutrition. The concept of “whole food” and the role of the food matrix in relation to healthy diets needs further clarification; the risk assessment/management of food additives also needs debate.

Series of five project update presentations

Almonds contain phytochemicals and nutrients that potentially have positive health benefits in relation to heart disease, diabetes and obesity. One important mechanism associated with these benefits is the bioaccessibility (release) of lipids from the almond tissue during mastication and digestion. Indeed, it has been reported that in human subjects on almond-rich diets, a significant proportion of lipid remains undigested, mainly caused by the entrapment of lipid by intact cell walls (dietary fiber). However, the bioaccessibility of almond lipid has not yet been quantified. We have studied this by performing a mastication trial in healthy human volunteers (n = 15). The particle size distribution of chewed almonds was measured using mechanical sieving and laser diffraction methods. Lipid release from ruptured almond cells was estimated using a theoretical model based on the particle size of the chewed almond tissue and the diameter of the lipid-rich cells. Our results showed that laser diffraction was the most reliable and efficient method. Lipid release from masticated almonds was estimated to be ~30% of total lipid and originated from the ruptured cells of the fractured tissue. This indicates that the majority of almond lipid is unavailable for the early stages of digestion. Further work will extend to studies of lipid release and digestion at more distal sites of the gastrointestinal tract

Introduction The proportion of lipid released during mastication of nuts is strongly influenced by particle size, due to natural encapsulation of the lipid by the walls of intact cells. The cell walls may act as barriers to digestion and may partially explain why nuts have reduced metabolizable energy versus the energy content predicted by Atwater factors. The lipid released from masticated nuts can be calculated using a mathematical model which has the cell diameter and the particle size distribution (PSD) of the bolus as variables. This study measured the cell size and PSD of four tree nuts (raw cashews, raw walnuts, roasted pistachios and raw Brazil nuts) to predict the lipid released due to their mastication. Objectives To predict the proportion of lipid released from masticated tree nuts, using measurements of cell size and PSD of masticated tree nuts (including cashews, walnuts, pistachios and Brazil nuts). To compare the results for these nuts to those already published for almonds. Method/Design Transverse and longitudinal sections were cut from each nut (including almonds) and micrographs processed using image analysis software to calculate the average cell diameter. Two randomized, un-blinded, cross-over trials were conducted. In each trial, 10 healthy volunteers attended two sessions, at which they chewed eight samples of a randomly allocated nut. For determination of PSDs, the expectorated boluses were sieved (2 boluses) or analyzed by laser diffraction (2 boluses). Initial lipid release was then predicted using the mathematical model. Results The cashew cells (34.3 μm) were smaller than the almond (45.1 μm), walnut (49.4 μm), pistachio (53.1 μm), and Brazil nut cells (60.8 μm). Laser diffraction showed that masticated nut boluses had median particle sizes (± SEM) which were smaller (cashews, 178 ± 12 μm; walnuts, 179 ± 8 μm; pistachios, 123 ± 10 μm; Brazil nuts, 145 ± 8 μm) than that for almonds measured previously (550 ± 18 μm). This results in higher predicted lipid release, calculated from the mathematical model, (mean, range) for cashews (12.3%, 8.7–16.3%), walnuts (14.5%, 12.0–18.0%), pistachios (11.0%, 8.7–13.0%) and Brazil nuts (14.4%, 11.9–17.3%) than for almonds (9.5%, 7.4–11.1%). Conclusions All of the five nuts had predicted lipid releases of less than 18%, which would be expected to attenuate postprandial lipemia and total nutrient availability relative to that expected due to their total lipid content. Due to their higher lipid content and predicted lipid release, walnuts and Brazil nuts (after mastication) are likely to release more fat on mastication than the other nuts tested.

Abstract 149/1077 (Poster)

Ingestion of different foods containing identical amounts of starch can result in very different postprandial rises in blood glucose and insulin concentrations. Limitation of the early rises in blood glucose and insulin levels seems to be beneficial to human health in the long term. Many studies of starch digestion in vitro are made to understand the molecular basis for differences in digestion rates in vivo to enable prediction of likely rates of digestion of particular starchy foods. Michaelis-Menten kinetics of starch digestibility provides estimates of available (digestible) substrate as starch samples are hydrothermally treated. Combined with studies of starch structure using calorimetry and FTIR spectroscopy, key features influencing rates of amylolysis were identified. Measurement of product formation during prolonged incubations of starch with α-amylase produces digestibility curves. Use of logarithm of slope (LOS) plots to analyse the curves by 1st order kinetics gave values for digestibility rate constants and the total digestible starch, C∞. An important conclusion is that contrary to many reports in the literature, cooked starches do not contain distinct fractions of rapidly and slowly digested material. These kinetic approaches have also been used in studies of plan-tencapsulated starch to understand how cell walls influence access of amylase to starch.

Seven project update posters