Within the active field of in vitro digestion in food research, the COST Action INFOGEST aimed to harmonizein vitro protocols simulating human digestion on the basis of physiologically inferred conditions. A harmonizedstatic in vitro digestion (IVD) method was recently published as a primary output from this network. To validatethis protocol, inter-laboratory trials were conducted within the INFOGEST network. A first study was performedusing skim milk powder (SMP) as a model food and served to compare the different in-house digestion protocolsused among the INFOGEST members. In a second inter-laboratory study applying the harmonized protocol, thedegree of consistency in protein hydrolysis was investigated. Analysis of the hydrolyzed proteins, after the gastric and intestinal phases, showed that caseins were mainly hydrolyzed during the gastric phase, whereas β-lactoglobulin was, as previously shown, resistant to pepsin. Moreover, generation of free amino acids occurredmainly during the intestinal phase.The study also showed that a few critical steps were responsible for the remaining inter-laboratory variability.The largest deviations arose from the determination of pepsin activity. Therefore, this step was further clarified,harmonized, and implemented in a third inter-laboratory study.The presentwork gives an overviewof all three inter-laboratory studies, showing that the IVD INFOGEST methodhas led to an increased consistency that enables a better comparability of in vitro digestion studies in the future

About 70∼100% of the Asian adult population is lactase non-persistent (LNP). The literature shows that many individuals with the LNP-genotype can consume ≤12g of lactose without experiencing gastrointestinal discomfort. Repetitive consumption of lactose may reduce intolerance symptoms via adaptation of the gut microbiota. This study aimed to assess the effects of daily consumption of incremental lactose doses on microbiota composition and function, and intolerance symptoms. Twenty-five healthy adults of Asian origin, carrying the LNP-genotype were included in this 12-week before-and-after intervention trial. Participants consumed gradually increasing lactose doses from 3 to 6g to 12g twice daily, each daily dose of 6, 12, or 24g being provided for 4 consecutive weeks. Participants handed in repeated stool samples and underwent a 25g lactose challenge hydrogen breath test (HBT) before and after the 12-week intervention. Daily gastrointestinal symptoms and total symptom scores (TSS) during the lactose challenge were recorded. A significant increase from 5.5±7.6% to 10.4±9.6% was observed in Bifidobacterium relative abundance after the intervention (p=0.009), accompanied by a two-fold increase (570±269U/g; p<0.001) in fecal β-galactosidase activity compared to baseline (272±158U/g). A 1.5-fold decrease (iAUC; p=0.01) in expired hydrogen was observed during the second HBT (38±35ppm·minutes), compared to the baseline HBT (57±38ppm·minutes). There was a non-significant decrease in TSS (10.6±8.3 before vs 8.1±7.2 after intervention; p=0.09). Daily consumption of lactose was well-tolerated, with mild to no gastrointestinal complaints reported during the intervention. Increased levels of Bifidobacterium indicate an adaptation of the gut microbiota upon repetitive consumption of incremental doses of lactose, which was well-tolerated as demonstrated by reduced expired hydrogen levels during the second 25g lactose HBT. Bifidobacteria metabolize lactose without gas production thereby potentially reducing intestinal gas formation in the gut of individuals with the LNP-genotype. This increased lactose tolerance possibly lifts the necessity to remove nutrient-rich dairy foods completely from the diet. The trial is registered at the International Clinical Trials Registry Platform: NL9516 The effect of dietary lactose in lactase non-persistent individuals on gut microbiota.

Exercise is one of the external factors associated with impairment of intestinal integrity, possibly leading to increased permeability and altered absorption. Here, we aimed to examine to what extent endurance exercise in the glycogen‐depleted state can affect intestinal permeability toward small molecules and protein‐derived peptides in relation to markers of intestinal function. Eleven well‐trained male volunteers (27 ± 4 years) ingested 40 g of casein protein and a lactulose/rhamnose (L/R) solution after an overnight fast in resting conditions (control) and after completing a dual – glycogen depletion and endurance – exercise protocol (first protocol execution). The entire procedure was repeated 1 week later (second protocol execution). Intestinal permeability was measured as L/R ratio in 5 h urine and 1 h plasma. Five‐hour urine excretion of betacasomorphin‐7 (BCM7), postprandial plasma amino acid levels, plasma fatty acid binding protein 2 (FABP‐2), serum pre‐haptoglobin 2 (preHP2), plasma glucagon‐like peptide 2 (GLP2), serum calprotectin, and dipeptidylpeptidase‐4 (DPP4) activity were studied as markers for excretion, intestinal functioning and recovery, inflammation, and BCM7 breakdown activity, respectively. BCM7 levels in urine were increased following the dual exercise protocol, in the first as well as the second protocol execution, whereas 1 h‐plasma L/R ratio was increased only following the first exercise protocol execution. FABP2, preHP2, and GLP2 were not changed after exercise, whereas calprotectin increased. Plasma citrulline levels following casein ingestion (iAUC) did not increase after exercise, as opposed to resting conditions. Endurance exercise in the glycogen depleted state resulted in a clear increase of BCM7 accumulation in urine, independent of DPP4 activity and intestinal permeability. Therefore, strenuous exercise could have an effect on the amount of food‐derived bioactive peptides crossing the epithelial barrier. The health consequence of increased passage needs more in depth studies.

Controlled studies on the effect of exercise on intestinal uptake of protein are scarce and underlying mechanisms largely unclear. We studied the uptake of the major allergen Ara h 6 following peanut consumption in an exercise model and compared this with changes in markers of intestinal permeability and integrity. Ten overnight-fasted healthy non-allergic men (n = 4) and women (n = 6) (23 ± 4 years) ingested 100 g of peanuts together with a lactulose/rhamnose (L/R) solution, followed by rest or by 60 min cycling at 70% of their maximal workload. Significantly higher, though variable, levels of Ara h 6 in serum were found during exercise compared to rest (Peak p = 0.03; area under the curve p = 0.006), with individual fold changes ranging from no increase to an increase of over 150-fold in the uptake of Ara h 6. Similarly, uptake of lactulose (2–18 fold change, p = 0.0009) and L/R ratios (0.4–7.9 fold change, p = 0.04) were significantly increased which indicates an increase in intestinal permeability. Intestinal permeability and uptake of Ara h 6 were strongly correlated (r = 0.77, p < 0.0001 for lactulose and Ara h 6). Endurance exercise after consumption may lead to increased paracellular intestinal uptake of food proteins.