Richard M. Faulks

The potential for the improvement of carotenoid levels in foods and the likely systemic effects

Carotenoids form one of the most important classes of plant pigments and play a crucial role in defining the quality parameters of fruit and vegetables. Their role in the plant is to act as accessory pigments for light harvesting and in the prevention of photo-oxidative damage, as well as acting as attractants for pollinators. Their function as antioxidants in the plant shows interesting parallels with their potential role as antioxidants in foods and humans. Carotenoids are products of the isoprenoid biosynthetic pathway. The enzymes leading to carotenoid biosynthesis have all been characterised, and more recently the genes encoding these enzymes have been cloned from bacteria, fungi and plants. New information on enzyme activities and the factors leading to the regulation of the pathway is reviewed. Vitamin A deficiency is a widespread problem in the developing world, causing blindness, particularly in the young. This has driven research into finding ways of introducing provitamin A carotenoids into staple crops, and this has recently been achieved in rice and canola through genetic manipulation. The fact that carotenoids show protective activity in vitro and in vivo against a variety of degenerative disease end points has also give impetus to studying whether increasing intakes of the commonly consumed carotenoids would have public health benefits in the developed world. Human intervention studies have been undertaken using supplements of β-carotene rather than utilising foods with enhanced carotenoid levels, but no potential benefit has been shown. Indeed, there is evidence of an increased health risk from the consumption of β-carotene supplements. These observations suggest that the threshold between the beneficial and adverse effects of some carotenoids is low and provides a strong stimulus to further understanding the functional effects of specific carotenoids. Specific needs for future research are identified in the review. © 2000 Society of Chemical Industry

In vitro digestion methods for assessing the effect of food structure on allergen breakdown

This article reviews the in vitro digestion models developed to assess the stability of food allergens during digestion. It is hypothesised that food allergens must exhibit sufficient gastro-intestinal stability to reach the intestinal mucosa where absorption and sensitisation (development of atopy) can occur. The investigation of stability of proteins within the gastrointestinal tract may provide prospective testing for allergenicity and could be a significant and valid parameter that distinguishes food allergens from nonallergens. Systematic evaluation of the stability of food allergens that are active via the gastrointestinal tract is currently tested in traditional pepsin digestibility models. The human gastrointestinal tract however is very complex and this article points out the importance of using physiologically relevant in vitro digestion systems for evaluating digestibility of allergens. This would involve the simulation of the stomach/small intestine environment (multi-phase models) with sequential addition of digestive enzymes, surfactants such as phospholipids and bile salts under physiological conditions, as well as the consideration of the effect of the food matrices on the allergen digestion.

Release of Protein, Lipid, and Vitamin E from Almond Seeds during Digestion

The evaluation of the bioaccessibility of almond nutrients is incomplete. However, it may have implications for the prevention and management of obesity and cardiovascular disease. This study quantified the release of lipid, protein, and vitamin E from almonds during digestion and determined the role played by cell walls in the bioaccessibility of intracellular nutrients. Natural almonds (NA), blanched almonds (BA), finely ground almonds (FG), and defatted finely ground almonds (DG) were digested in vitro under simulated gastric and gastric followed by duodenal conditions. FG were the most digestible with 39, 45, and 44% of lipid, vitamin E, and protein released after duodenal digestion, respectively. Consistent with longer residence time in the gut, preliminary in vivo studies showed higher percentages of nutrient release, and microscopic examination of digested almond tissue demonstrated cell wall swelling. Bioaccessibility is improved by increased residence time in the gut and is regulated by almond cell walls.

Effect of intragastric acid stability of fat emulsions on gastric emptying, plasma lipid profile and postprandial satiety

Fat is often included in common foods as an emulsion of dispersed oil droplets to enhance the organoleptic quality and stability. The intragastric acid stability of emulsified fat may impact on gastric emptying, satiety and plasma lipid absorption. The aim of the present study was to investigate whether, compared with an acid-unstable emulsion, an acid-stable fat emulsion would empty from the stomach more slowly, cause more rapid plasma lipid absorption and cause greater satiety. Eleven healthy male volunteers received on two separate occasions 500 ml of 15 % (w/w) [13C]palmitate-enriched olive oil-in-water emulsion meals which were either stable or unstable in the acid gastric environment. MRI was used to measure gastric emptying and the intragastric oil fraction of the meals. Blood sampling was used to measure plasma lipids and visual analogue scales were used to assess satiety. The acid-unstable fat emulsion broke and rapidly layered in the stomach. Gastric emptying of meal volume was slower for the acid-stable fat emulsion (P < 0.0001; two-way ANOVA). The rate of energy delivery of fat from the stomach to the duodenum was not different up to t = 110 min. The acid-stable emulsion induced increased fullness (P < 0.05), decreased hunger (P < 0.0002), decreased appetite (P < 0.0001) and increased the concentration of palmitic acid tracer in the chylomicron fraction (P < 0.04). This shows that it is possible to delay gastric emptying and increase satiety by stabilising the intragastric distribution of fat emulsions against the gastric acid environment. This could have implications for the design of novel foods.

Modulating pancreatic lipase activity with galactolipids: effects of emulsion interfacial composition

It is widely known that the interfacial quality of lipid emulsion droplets influences the rate and extent of lipolysis. The aim of this work was to investigate the effect of two galactolipids, monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), adsorbed at the interface on in vitro digestibility of olive oil by porcine pancreatic lipase. The experiments were performed under simulated duodenal conditions in the presence of phosphatidylcholine (lecithin) and bile salts. It was found that emulsions prepared with DGDG had a longer lag phase prior to lipase activation with a decrease in lipolysis rate. In contrast, no inhibitory effect on lipase kinetics was observed in emulsions prepared with MGDG. We postulated that the larger headgroup and more tightly packed molecular organization of DGDG at the interface gave rise to steric hindrance that retarded colipase and lipase adsorption onto the substrate surfaces and hence delayed and reduced lipolysis. It was noted that the lag phase and lipolysis rate strongly depended on the DGDG/lecithin molar ratio in the systems: the higher the molar ratio, the longer the lag phase followed by a reduced lipolysis rate. The ability of DGDG to inhibit bile salt adsorption/displacement was also investigated. The results showed that bile salts did not completely displace DGDG from the interface, explaining the reason why DGDG still possessed inhibitory activity even in the presence of bile salts at a physiological relevant concentration. The results provide interesting insights into the influence of the galactolipid headgroup and lecithin on the emulsion interfacial quality which in turn regulates the lipolysis. The findings potentially could lead to the production of generic foods and drugs designed for regulating dietary fat absorption in the prevention and treatment of obesity and related disorders.

Solubilization of carotenoids from carrot juice and spinach in lipid phases: II. Modeling the duodenal environment

Our understanding of the factors determining the bioavailability of carotenoids from fruits and vegetables is poor. The apolar nature of carotenoids precludes their simple diffusion from the food structure to the absorption site at the enterocyte. Therefore, there is interest in the potential pathways for solubilization in the gut before absorption. We have studied the transfer of carotenoids from carrot juice and homogenized spinach into lipid phases that mimic the intestinal lumen at the start of digestion. In this paper we report on their transfer into olive oil under conditions pertaining to the gastric environment. A comparison between preparations of raw spinach and of carrot, in which the intact cells have been largely broken, suggests that the membrane-bound carotenoids of spinach are more resistant to transfer than the crystalline carotenoids of carrot. Lowering the pH and pepsin treatment enhance the transfer from raw vegetables. The process of blanching and freezing spinach destroys the chloroplast ultrastructure and leads to (i) a substantial increase in transfer of the carotenoids to oil and (ii) an attenuation or reversal of the enhancement of transfer seen with reduced pH or with pepsin treatment. Similar effects are seen after blanching carrot juice. Our results show that removal of soluble protein and denaturation of membrane proteins enhances the partition of carotenoids into oil. For both vegetables there is no evidence of preference in the extent of transfer of one carotenoid over another. This suggests that partitioning into oil under gastric conditions is not the stage of digestion that could lead to differences in carotenoid bioavailability.

Effect of carrot (Daucus carota) microstructure on carotene bioaccessibilty in the upper gastrointestinal tract. 1. in vitro simulations of carrot digestion

Studies investigating carotene bioaccessibility (release from the food matrix to a solubilized form) directly from plant material during the process of digestion are scarce, mainly due to the difficulties associated with obtaining such material. Therefore, this paper examines the relationship between tissue microstructure and carotene bioaccessibility using an in vitro digestion model. Dietary oil provides a pool for the initial solubilization. Therefore, carotene partitioning into an emulsified oil phase was assessed using raw carrot tissue and carrot tissue subjected to various degrees of heating and particle size reduction and, in all cases, was found to be greatly reduced compared with juiced carrot. Carotene bioaccessibility was found to be greater from raw tissues than heated tissues of the same size. This is because heating increases the propensity for intact cells to separate, effectively encapsulating the carotene. Although the gross structure of the tissues was found to be relatively unaffected by in vitro digestion, at the cellular level some cell-wall swelling and cell death were observed, particularly close to the surfaces of the tissue. This study suggests that cell-wall rupture prior to digestion is an absolute requirement for carotene bioaccessibility in the upper intestine and that heating does not enhance carotene release from intact cells.

Survival of Lactobacillus rhamnosus strains in the upper gastrointestinal tract

In the present study six probiotic Lactobacillus rhamnosus strains were investigated for their ability to survive in the human upper gastrointestinal tract through a dynamic gastric model of digestion. MRS broth was used as delivery vehicle and survival was investigated during in vitro gastric and gastric plus duodenal digestion. Results highlighted that all tested strains showed good survival rate during both gastric and duodenal digestion. In particular, three strains exhibited a great survival showing a recovery percentage in the range between 117 and 276%. In agreement with survival data, high lactic acid production was detected for all strains, confirming their metabolic activity during digestion.

Bioaccessibility of pistachio polyphenols, xanthophylls, and tocopherols during simulated human digestion

OBJECTIVE The bioaccessibility of bioactives from pistachios has not been previously evaluated. In the present study we quantified the release of polyphenols, xanthophylls (lutein), and tocopherols from pistachios (raw pistachios, roasted salted pistachios, and muffins made with raw pistachios) during simulated human digestion. METHODS A dynamic gastric model of digestion that provides a realistic and predictive simulation of the physical and chemical processing and accurately mimics the residence time and the luminal environment within the human stomach was used for the digestion studies. RESULTS More than 90% of the polyphenols were released in the gastric compartment, with virtually total release in the duodenal phase. No significant differences were observed between raw shelled and roasted salted pistachio. The presence of a food matrix (muffin) decreased the bioaccessibility of protocatechuic acid (78%) and luteolin (36%). Almost 100% bioaccessibility of lutein and tocopherols was found after duodenal digestion, with no difference among the three samples. CONCLUSION The rapid release of the assayed bioactives in the stomach maximizes the potential for absorption in the duodenum and contributes to the beneficial relation between pistachio consumption and health-related outcomes.

Simultaneous time-varying systemic appearance of oral and hepatic glucose in adults monitored with stable isotopes

The rates (and extent) of appearance of glucose in arterialized plasma from an oral glucose load and from liver (RaO, RaH) can be estimated in humans using radioisotopes, but estimates vary among laboratories. We investigated the use of stable isotopes and undertook 22 primed intravenous infusions of D-[6,6-2H2]glucose with an oral load including D-[13C6]glucose in healthy humans. The effective glucose pool volume (VS) had a lower limit of 230 ml/kg body weight (cf. 130 ml/kg commonly assumed). This VS in Steeles one-compartment model of glucose kinetics gave a systemic appearance from a 50-g oral glucose load per 70 kg body weight of 96 +/- 3% of that ingested, which compared with a theoretical value of approximately 95%. Maris two-compartment model gave 100 +/- 3%. The two models gave practically identical RaO and RaH at each point in time and a plateau in the cumulative RaO when absorption was complete. Less than 3% of 13C was recycled to [13C3]glucose, suggesting that recycling errors were practically negligible in this study. Causes of variation among laboratories are identified. We conclude that stable isotopes provide a reliable and safe alternative to radioactive isotopes in these studies.The rates (and extent) of appearance of glucose in arterialized plasma from an oral glucose load and from liver (RaO, RaH) can be estimated in humans using radioisotopes, but estimates vary among laboratories. We investigated the use of stable isotopes and undertook 22 primed intravenous infusions ofd-[6,6-2H2]glucose with an oral load includingd-[13C6]glucose in healthy humans. The effective glucose pool volume (VS) had a lower limit of 230 ml/kg body weight (cf. 130 ml/kg commonly assumed). This VS in Steeles one-compartment model of glucose kinetics gave a systemic appearance from a 50-g oral glucose load per 70 kg body weight of 96 ± 3% of that ingested, which compared with a theoretical value of ∼95%. Maris two-compartment model gave 100 ± 3%. The two models gave practically identical RaO and RaH at each point in time and a plateau in the cumulative RaO when absorption was complete. Less than 3% of13C was recycled to [13C3]glucose, suggesting that recycling errors were practically negligible in this study. Causes of variation among laboratories are identified. We conclude that stable isotopes provide a reliable and safe alternative to radioactive isotopes in these studies.