Heat treatment and homogenization influence the gastric digestion of bovine milk protein in growing pigs as an adult human model.

BACKGROUND: Bovine milk processing influences the structure of the curd formed during gastric digestion, which may alter gastric protein hydrolysis and impact amino acid (AA) release into the small intestine. OBJECTIVE: This study aimed to determine the influence of heat treatment and homogenization on the gastric protein digestion and amino acid emptying of bovine milk. METHODS: Nine-week-old pigs (n=144) consumed either raw, pasteurized non-homogenized (PNH), pasteurized-homogenized (PH), or ultra-high-temperature homogenized (UHT) bovine milk for ten days. On day 11, fasted pigs received the milk treatment (500 mL) before gastric contents were collected at 0, 20, 60, 120, 180, and 300 min postprandially. The apparent degree of gastric protein hydrolysis (based on the release of free amino groups), apparent gastric disappearance of individual proteins (based on SDS-PAGE gel band intensity), and the gastric emptying of digested protein and AA were determined. RESULTS: During the first 60 min, the rate of apparent gastric protein hydrolysis was fastest for pigs fed UHT milk (0.29%/min vs on average 0.07%/min for pigs fed raw, PNH, and PH milk). Differences in the apparent degree of gastric protein hydrolysis and emptying were reflected in the rate of digested protein entering the small intestine. The AA gastric emptying half-time was generally shorter for pigs fed PH and UHT milk than pigs fed raw and PNH milk. For example, the gastric release of total essential AA was more than twofold faster (P<0.01) for pigs fed PH or UHT milk compared to pigs fed raw or PNH milk (i.e., homogenized vs non-homogenized milk). CONCLUSIONS: Heat treatment and homogenization increased the apparent gastric degree of protein hydrolysis and the release of digested protein into the small intestine. However, the rate of AA entering the small intestine was mainly increased by homogenization.

Effects of Green and Gold Kiwifruit Varieties on Antioxidant Neuroprotective Potential in Pigs as a Model for Human Adults.

Kiwifruit (KF) has shown neuroprotective potential in cell-based and rodent models by augmenting the capacity of endogenous antioxidant systems. This study aimed to determine whether KF consumption modulates the antioxidant capacity of plasma and brain tissue in growing pigs. Eighteen male pigs were divided equally into three groups: (1) bread, (2) bread + Actinidia deliciosa cv. 'Hayward' (green-fleshed), and (3) bread + A. chinensis cv. 'Hort16A' (yellow-fleshed). Following consumption of the diets for eight days, plasma and brain tissue (brain stem, corpus striatum, hippocampus, and prefrontal cortex) were collected and measured for biomarkers of antioxidant capacity, enzyme activity, and protein expression assessments. Green KF significantly increased ferric-reducing antioxidant potential (FRAP) in plasma and all brain regions compared with the bread-only diet. Gold KF increased plasma ascorbate concentration and trended towards reducing acetylcholinesterase activity in the brain compared with the bread-only diet. Pearson correlation analysis revealed a significant positive correlation between FRAP in the brain stem, prefrontal cortex, and hippocampus with the total polyphenol concentration of dietary interventions. These findings provide exploratory evidence for the benefits of KF constituents in augmenting the brain's antioxidant capacity that may support neurological homeostasis during oxidative stress.

Validation of a Combined In Vivo/In Vitro Ileal Fermentation Assay in the Growing Pig to be Used as a Model for Adult Humans.

BACKGROUND: An in vivo/in vitro ileal fermentation assay using growing pigs has been developed but not yet formally validated. OBJECTIVES: This study aimed to validate the in vivo/in vitro ileal fermentation assay by comparing in vitro fermentation values with those obtained in vivo in growing pigs. The effect of raising pigs under different environmental conditions was also investigated. METHODS: Thirty piglets (1.59 ± 0.31 kg body weight, mean ± standard deviation) were subjected to 1 of 3 treatments: artificially reared (AR) (nonfarm, laboratory housing conditions) from postnatal day (PND) 7 (AR group), inoculated orally with human infant fecal extracts from birth until PND 8 and AR (AR+ group), or conventionally reared on a farm (control group). Starting at PND 7, the AR and AR+ pigs received human infant formula for 3 wk, followed by a human-type diet for 5 wk. Control pigs were weaned on the farm and, on PND 63, relocated to the laboratory animal facility. From PND 63, all pigs received a human-type diet. On PND 78, pigs were killed, after which ileal digesta were collected to perform an in vitro ileal fermentation (in vitro organic matter [OM] fermentability and organic acid production) and to determine digesta microbial composition and dietary OM fermentability in vivo. RESULTS: The rearing regimen resulted in only a few differences in ileal microbial taxonomic composition. The rearing regimen generally did not affect the in vitro production of individual organic acids. The in vivo and in vitro OM fermentability of proximal ileal digesta (19.7 ± 2.04%; mean ± SEM) was similar (P > 0.05) for the AR and control pigs but not for the AR+ pigs. CONCLUSIONS: The control-rearing regimen was preferred over AR or AR+ because of ease of implementation. The in vitro ileal fermentation assay accurately predicted the in vivo OM fermentability.

The kinetics of amino acid disappearance in the small intestine is related to the extent of amino acids absorbed in growing pigs.

This study evaluated the importance of a correction for amino acids (AA) released into the hindgut on a measure of AA absorption kinetics and tested whether AA absorption kinetics are related to the extent of AA absorption using the growing pig as a model for humans. Thirty-six nine-week-old pigs (22·3 kg) received a diet containing whey protein as the sole protein source for 8 d. Pigs received their last meal containing the indigestible marker titanium dioxide before being euthanised at 1, 2, 3, 4, 6 and 12 h post-feeding. The entire content of each gastrointestinal tract (GIT) region was collected to determine AA released into the hindgut, and the kinetics and extent of AA absorption (uncorrected and corrected for AA entering the hindgut). Amounts of AA released into the hindgut increased over time (e.g. 33 and 180 mg of Glu for 4 and 6 h post-feeding). The corrected apparent amount of each AA absorbed from the GIT lumen after 4 h post-feeding was generally lower (P ≤ 0·05) than the uncorrected counterpart. Differences in both the kinetics and extent of AA absorption were observed across AA. For example, the time to reach half of the apparent AA absorption (T50) was 1·5 and 3·4 h for Met and Arg, respectively, whereas their extent of apparent absorption was 93 and 73 %. Negative correlations between parameters related to kinetics and the extent of apparent absorption were observed (e.g. for T50 r = -0·81; P < 0·001). The kinetics of AA absorption is related to the extent of AA absorption.

The probiotic Lacticaseibacillus rhamnosus HN001 influences the architecture and gene expression of small intestine tissue in a piglet model.

This study investigated the effects of Lacticaseibacillus rhamnosus HN001 supplementation on the architecture and gene expression in small intestinal tissues of piglets used as an animal model for infant humans. Twenty-four 10-d-old entire male piglets (4·3 (sd 0·59) kg body weight) were fed an infant formula (IF) (control) or IF supplemented with 1·3 × 105 (low dose) or 7·9 × 106 (high dose) colony-forming units HN001 per ml of reconstituted formula (n 8 piglets/treatment). After 24 d, piglets were euthanised. Samples were collected to analyse the histology and gene expression (RNAseq and qPCR) in the jejunal and ileal tissues, blood cytokine concentrations, and blood and faecal calprotectin concentrations. HN001 consumption altered (false discovery rate < 0·05) gene expression (RNAseq) in jejunal tissues but not in ileal tissues. The number of ileal goblet cells and crypt surface area increased quadratically (P < 0·05) as dietary HN001 levels increased, but no increase was observed in the jejunal tissues. Similarly, blood plasma concentrations of IL-10 and calprotectin increased linearly (P < 0·05) as dietary HN001 levels increased. In conclusion, supplementation of IF with HN001 affected the architecture and gene expression of small intestine tissue, blood cytokine concentration and frequencies, and blood calprotectin concentrations, indicating that HN001 modulated small intestinal tissue maturation and immunity in the piglet model.

Metabolite profiling of peripheral blood plasma in pigs in early postnatal life fed whole bovine, caprine or ovine milk.

Ruminants' milk is commonly used for supplying nutrients to infants when breast milk is unavailable or limited. Previous studies have highlighted the differences between ruminants' milk composition, digestion, absorption, and fermentation. However, whether consuming different ruminants' milk impact the appearance of the circulatory blood metabolites in the early postnatal life is not well understood. The analysis conducted here aimed to determine the effect of feeding exclusively whole milk from bovine, caprine or ovine species to pigs, approximately 7 days-old for 15 days, on circulatory blood plasma metabolites. Relative intensities of plasma metabolites were detected using a liquid chromatography-mass spectrometry based metabolomic approach. Seven polar and 83 non-polar (lipids) metabolites in plasma were significantly different (false discovery rate < 0.05) between milk treatments. These included polar metabolites involved in amino acid metabolism and lipids belonging to phosphatidylcholine, lysophosphatidylcholine, sphingomyelin, and triglycerides. Compared to the caprine or bovine milk group, the relative intensities of polar metabolites and unsaturated triglycerides were higher in the peripheral circulation of the ovine milk group. In contrast, relative intensities of saturated triglycerides and phosphatidylcholine were higher in the bovine milk group compared to the ovine or caprine milk group. In addition, correlations were identified between amino acid and lipid intake and their appearance in peripheral blood circulation. The results highlighted that consuming different ruminants' milk influences the plasma appearance of metabolites, especially lipids, that may contribute to early postnatal life development in pigs.

Effect of Heat Treatment on Protein Self-Digestion in Ruminants' Milk.

This study investigated whether heat treatments (raw, 63 °C for 30 min, and 85 °C for 5 min) affect protein hydrolysis by endogenous enzymes in the milk of ruminants (bovine, ovine, and caprine) using a self-digestion model. Self-digestion consisted of the incubation for six hours at 37 °C of the ruminants' milk. Free amino group concentration was measured by the o-phthaldialdehyde method, and peptide sequences were identified by chromatography-mass spectrometry. Results showed that heat treatments prior to self-digestion decreased the free NH2 by 59% in bovine milk heated at 85 °C/5 min, and by 44 and 53% in caprine milk heated at 63 °C/30 min and 85 °C/5 min, respectively. However, after self-digestion, only new free amino groups were observed for the raw and heated at 63 °C/30 min milk. ß-Casein was the most cleaved protein in the raw and heated at 63 °C/30 min bovine milk. A similar trend was observed in raw ovine and caprine milk. Self-digestion increased 6.8-fold the potential antithrombin peptides in the bovine milk heated at 63 °C/30 min. Enhancing bioactive peptide abundance through self-digestion has potential applications in the industry for functional products. Overall, heat treatments affected the free amino groups according to the species and heat treatment applied, which was reflected in the varying degrees of cleaved peptide bonds and peptides released during self-digestion.

Differences in small intestinal apparent amino acid digestibility of raw bovine, caprine, and ovine milk are explained by gastric amino acid retention in piglets as an infant model.

Background: The rate of stomach emptying of milk from different ruminant species differs, suggesting that the small intestinal digestibility of nutrients could also differ across these milk types. Objective: To determine the small intestinal amino acid (AA) digestibility of raw bovine, caprine, and ovine milk in the piglet as an animal model for the infant. Methods: Seven-day-old piglets (n = 12) consumed either bovine, caprine, or ovine milk diets for 15 days (n = 4 piglets/milk). On day 15, fasted piglets received a single meal of fresh raw milk normalized for protein content and containing the indigestible marker titanium dioxide. Entire gastrointestinal tract contents were collected at 210 min postprandially. Apparent AA digestibility (disappearance) in different regions of the small intestine was determined. Results: On average, 35% of the dietary AAs were apparently taken up in the small intestine during the first 210 min post-feeding, with 67% of the AA digestibility occurring in the first quarter (p ≤ 0.05) and 33% in the subsequent two quarters. Overall, except for isoleucine, valine, phenylalanine, and tyrosine, the small intestinal apparent digestibility of all AAs at 210 min postprandially in piglets fed ovine milk was, on average, 29% higher (p ≤ 0.05) than for those fed bovine milk. Except for lysine, there was no difference in the apparent digestibility (p > 0.05) of any AAs between piglets fed caprine milk or ovine milk. The apparent digestibility of alanine was higher (p ≤ 0.05) in piglets fed caprine milk than those fed bovine milk. When apparent digestibility was corrected for gastric AA retention, only small differences in the small intestinal apparent digestibility of AAs were observed across milk types. Conclusion: Bovine, caprine and ovine milk had different apparent small intestinal AA digestibility at 210 min postprandially. When corrected for gastric AA retention, the differences in apparent digestibility across species largely disappeared. The apparent AA digestibility differed across small intestinal locations.

Type and Amount of Dietary Fiber Influence the Hindgut Synthesis of Organic Acids from Fermentable Material of Both Total and Nondietary Origin in a Pig Model of the Adult Human.

BACKGROUND: Organic acid synthesis by the hindgut microbiota is commonly believed to be mainly of fermentable material of dietary origin. OBJECTIVE: This study aimed to determine the hindgut organic acid synthesis from fermentable material of dietary (mainly fiber) or nondietary origin for different types and amounts of dietary fiber in growing pigs used as a model for adult humans. METHOD: Seven fiber-containing diets were formulated: 4 fiber types (cellulose, gum acacia, oligofructose, and pectin) at 6% of the diet and 3 (gum acacia, oligofructose, and pectin) at 3% as the sole fiber source. Ileal cannulated female pigs (n = 14; Landrace/Large white) were fed the fiber-containing diets (n = 6 pigs/diet) for 11 days (fiber phase) followed by 3 days on a fiber-free diet (fiber-free phase), using a replicated Youden square. Ileal digesta for each phase were collected and fermented in vitro with a pooled fecal microbial inoculum prepared from feces collected during the fiber phase to determine the organic acids synthesized from fermentable material of dietary (fiber phase) and nondietary (fiber-free phase) origins. RESULTS: The total amount of each individual organic acid synthesized during in vitro hindgut fermentation differed (P ≤ 0.05) across the types and amounts of dietary fiber intake. For example, the amount of acetate was 3.6-fold higher (P ≤ 0.05) for pigs fed the 6% pectin-containing diet than those fed the 6% oligofructose-containing diet. The nondietary substrate contributed between 36% (hexanoate) and 70% (succinate) to the total hindgut organic acid synthesis. The adaptation to the different fiber-containing diets led to different amounts of some organic acids of nondietary origin. CONCLUSIONS: The total amount of organic acids synthesized in the hindgut by the resident microbes is influenced by the type and amount of dietary fiber consumed. This study quantifies the interaction between both dietary and nondietary fermentable materials in hindgut fermentation.

Biotransformation of Rutin in In Vitro Porcine Ileal and Colonic Fermentation Models.

Quercetin, a polyphenol antioxidant, is widely distributed in food in the form of glycoside rutin, which is not readily absorbed in the gastrointestinal tract. The microbiota of the colon is known to biotransform rutin, generating quercetin aglycones that can be absorbed. We investigated the role of the ileal and colonic microbiota in rutin biotransformation using established in vitro fermentation models. Overall, a higher rate of rutin biotransformation was observed during colonic fermentation compared with ileal fermentation. The colonic microbiome showed higher potential for rutin conversion to quercetin through an increased abundance of α-rhamnosidase- and ß-glucosidase-encoding genes compared to the ileal microbiome. Nonetheless, rutin metabolism occurred rapidly during ileal fermentation (∼20% rutin disappearance after 1 h). The appearance of quercetin varied depending on the ileal inoculum and correlated with an increased abundance of Firmicutes, suggesting that quercetin absorption could be improved via modulation of the ileal microbiota.

The impact of heating and drying on protease activities of ruminant milk before and after in vitro infant digestion.

This study investigated the effect of heating (63°C/30 min or 75°C/15 s) and drying (spray-drying or freeze-drying) on plasmin, cathepsin D, and elastase activities in bovine, ovine, and caprine milk, compared to non-dried raw milk counterparts. Protease activities and protein hydrolysis were assessed before and after in vitro infant digestion with or without gastric and pancreatic enzymes. At 75°C/15 s, plasmin activity in caprine and ovine milk decreased (69-75%, p<0.05), while cathepsin D activity in spray-dried bovine milk heated increased (2.8-fold, p<0.05). Plasmin and cathepsin D activities increased (<1.2-fold, p<0.05) after in vitro digestion with pancreatin, regardless of milk species. Endogenous milk enzymes hydrolyzed more proteins than gastric enzymes during gastric digestion and contributed to small intestinal digestion. In summary, milk proteases remained active after processing with effects dependent on the species of milk, and they contributed to in vitro protein hydrolysis in the stomach and small intestine.

In Vitro Ileal Fermentation is Affected More by the Fiber Source Fermented than the Ileal Microbial Composition in Growing Pigs.

Background: The fermentation of undigested material in the ileum is quantitatively important. However, the respective contributions of the microbial composition and the substrate to ileal fermentation are unclear. Objective: This aim was to investigate the contribution of microbial composition and fiber source to in vitro ileal fermentation outcomes. Methods: Thirteen ileal cannulated female pigs (Landrace/Large White; 9-wk-old; 30.5 kg body weight) were given diets containing black beans, wheat bread, chickpeas, peanuts, pigeon peas, sorghum, or wheat bran as the sole protein source for 7 d (100 g protein/kg dry matter diet). On day 7, ileal digesta were collected and stored at -80°C for microbial analysis and in vitro fermentation. For each diet, a pooled ileal inoculum was prepared to ferment different fiber sources (cellulose, pectin, arabinogalactan, inulin, fructooligosaccharides, and resistant starch) for 2 h at 37°C. Organic matter fermentability and organic acid production were determined following in vitro fermentation. Data were analyzed using a 2-way ANOVA (inoculum × fiber). Results: Forty-five percent of the identified genera in the digesta differed across diets. For instance, the number of Lactococcus was 115-fold greater (P ≤ 0.05) in the digesta of pigs fed the pigeon pea diet than for pigs fed the wheat bran diet. For both in vitro organic matter fermentability and organic acid production, there were significant (P ≤ 0.05) interactions between the inoculum and the fiber source. For instance, pectin and resistant starch resulted in 1.6- to 31-fold more (P ≤ 0.05) lactic acid production when fermented by the pigeon pea inoculum than other inocula. For specific fiber sources, statistically significant correlations were found between the number of bacteria from certain members of the ileal microbial community and fermentation outcomes. Conclusions: Both the fiber source fermented and the ileal microbial composition of the growing pig affected in vitro fermentation; however, the effect of the fiber source was predominant.Curr Dev Nutr 2023;x:xx.

Available Lysine in Foods as Determined in Adult Ileostomates.

BACKGROUND: When foods are subjected to heat and pressure, a proportion of lysine is structurally altered and some will revert to lysine because of acid hydrolysis during amino acid analysis. Altered lysine molecules may be partly absorbed but are not utilized post-absorption. OBJECTIVES: A guanidination-based bioassay has been developed to determine true ileal digestible reactive lysine but it was only used in animal models (pig and rat). The objective of this study was to apply the assay and determine whether there is a difference between true ileal digestible total lysine and true ileal digestible reactive lysine in adult human ileostomates. METHODS: Six cooked or processed foods were analyzed for total lysine and reactive lysine. Six adults with a fully functioning ileostomy (4 women and 2 men; age range: 41-70 y; BMI: 20.8-28.1) participated. Foods for which total lysine > reactive lysine (cooked black beans, toasted wheat bread, and processed wheat bran), as well as a protein-free diet, were consumed by the ileostomates (n = 5 to 8; test food meals contained 25 g protein) and ileal digesta was collected. Each food was ingested twice by each participant, and the digesta was pooled. The order of foods for each participant was determined according to a Youden square. True ileal digestible total lysine and true ileal digestible reactive lysine were determined and a 2-way ANOVA model was used to analyze data. RESULTS: True ileal digestible reactive lysine was significantly lower than true ileal digestible total lysine for cooked black beans, toasted wheat bread, and processed wheat bran by 89%, 55%, and 85%, respectively (P< 0.05). CONCLUSIONS: True ileal digestible reactive lysine was lower than true ileal digestible total lysine, similar to that previously reported in pigs and rats, demonstrating the importance of determining the true ileal digestible reactive lysine contents of processed foods.

Ileal Digestibility of Nitrogen and Amino Acids in Human Milk and an Infant Formula as Determined in Neonatal Minipiglets.

BACKGROUND: Infant formula (IF) has to provide at least the same amount of amino acids (AAs) as human milk (HM). AA digestibility in HM and IF was not studied extensively, with no data available for tryptophan digestibility. OBJECTIVES: The present study aimed to measure the true ileal digestibility (TID) of total nitrogen and AAs in HM and IF to estimate AA bioavailability using Yucatan mini-piglets as an infant model. METHODS: Twenty-four 19-day-old piglets (males and females) received either HM or IF for 6 days or a protein-free diet for 3 days, with cobalt-EDTA as an indigestible marker. Diets were fed hourly over 6 h before euthanasia and digesta collection. Total N, AA, and marker contents in diets and digesta were measured to determine the TID. Unidimensional statistical analyses were conducted. RESULTS: Dietary N content was not different between HM and IF, while true protein was lower in HM (-4 g/L) due to a 7-fold higher non-protein N content in HM. The TID of total N was lower (P < 0.001) for HM (91.3 ± 1.24%) than for IF (98.0 ± 0.810%), while the TID of amino acid nitrogen (AAN) was not different (average of 97.4 ± 0.655%, P = 0.272). HM and IF had similar (P > 0.05) TID for most of the AAs including tryptophan (96.7 ± 0.950%, P = 0.079), except for some AAs (lysine, phenylalanine, threonine, valine, alanine, proline, and serine), with small significant difference (P < 0.05). The first limiting AA was the aromatic AAs, and the digestible indispensable AA score (DIAAS) was higher for HM (DIAASHM = 101) than for IF (DIAASIF = 83). CONCLUSION: HM, compared to IF, had a lower TID for total N only, whereas the TID of AAN and most AAs, including Trp, was high and similar. A larger proportion of non-protein N is transferred to the microbiota with HM, which is of physiological relevance, although this fraction is poorly considered for IF manufacturing.

Effects of the maize-derived protein zein, and the milk proteins casein, whey, and α-lactalbumin, on subjective measures of satiety and food intake in normal-weight young men.

Protein is considered to be the most satiating food macronutrient and the satiating effect may be dependent on the source of the protein. The maize-derived protein zein and milk protein casein have been shown previously to lower stomach emptying rate more than dairy whey protein, but the effect of zein on satiety has not been evaluated. The objective was to compare the satiating effects of zein and casein, with whey protein and its protein component α-lactalbumin. The study was a randomised crossover design with thirteen normal-weight men (mean age 27.8 years and mean BMI 24.4 kg/m2) consuming isoenergetic (∼4000 kJ, ∼990 kcal) preload mixed meals enriched with Zein, Casein, whey protein isolate (Whey), α-lactalbumin (ALac), or maltodextrin carbohydrate (Carb). Consumption of an ad libitum standardised test meal of chicken fried rice and water provided 360 min following ingestion of the preload meal was measured, and subjective feelings of appetite (hunger, fullness, desire to eat, and prospective food consumption) were assessed using 100-mm visual analogue scales (VAS). There were no differences among the five preload mixed meals in the amount of chicken fried rice consumed at the ad libitum test meal (mean ± sem: 531.6 ± 35.0 g, p = 0.47) or total (preload + test meal) energy intakes (mean ± sem: 5780.5 ± 146.0 kJ, p = 0.29). The subjective VAS appetite ratings and total area under the curve responses for hunger, fullness, desire to eat, and prospective food consumption, were not different following consumption of all five preload mixed meals (p > 0.05). The findings indicate that the effects of zein and casein on satiety were not different from the satiating effects of whey protein and α-lactalbumin.

A Method Using Longitudinal Laboratory Data to Predict Future Intestinal Complication in Patients with Crohn's Disease.

BACKGROUND: Stenosis, fistulization, and perforation of the bowel are severe outcomes which can occur in patients with Crohn's disease. Accurate prediction of these events may enable clinicians to alter treatment strategies and avoid these outcomes. AIMS: To study the correlation between longitudinal laboratory testing and subsequent intestinal complications in patients with Crohn's disease. METHODS: An observational cohort of patients with Crohn's disease at a single center were analyzed between 01/01/1994 and 06/30/2016. A complication was defined as the development of an intestinal fistula, stenosis, or perforation. Exploratory analysis using Cox regression was performed to select the best statistical method to represent longitudinal laboratory data. Cox regression was used to identify laboratory variables independently associated with the development of a subsequent complication. A clinical scoring tool was designed. RESULTS: In 246 patients observed over a median of 5.72 years, 134 complications occurred. Minimum or maximum value in a preceding window period of one year was most strongly associated with subsequent complication. A Longitudinal Laboratory score of ≥ 2 (maximum albumin level < 39 g/L = 1, maximum mean cell volume < 88 fL = 1, minimum platelet count > 355 × 109/L = 1, minimum C reactive protein > 5 mg/L = 1) was 62% sensitive and 91% specific in identifying patients who develop a subsequent complication. CONCLUSION: A consistent reduction in serum albumin and mean cell volume, and a consistent increase in platelet count and C reactive protein were associated with a subsequent complication in patients with Crohn's disease. Longitudinal laboratory tests may be used as described in this paper to provide a rational for earlier escalation of therapy.

Infant nutrition affects the microbiota-gut-brain axis: Comparison of human milk vs. infant formula feeding in the piglet model.

Early nutrition plays a dominant role in infant development and health. It is now understood that the infant diet impacts the gut microbiota and its relationship with gut function and brain development. However, its impact on the microbiota-gut-brain axis has not been studied in an integrative way. The objective here was to evaluate the effects of human milk (HM) or cow's milk based infant formula (IF) on the relationships between gut microbiota and the collective host intestinal-brain axis. Eighteen 10-day-old Yucatan mini-piglets were fed with HM or IF. Intestinal and fecal microbiota composition, intestinal phenotypic parameters, and the expression of genes involved in several gut and brain functions were determined. Unidimensional analyses were performed, followed by multifactorial analyses to evaluate the relationships among all the variables across the microbiota-gut-brain axis. Compared to IF, HM decreased the α-diversity of colonic and fecal microbiota and modified their composition. Piglets fed HM had a significantly higher ileal and colonic paracellular permeability assessed by ex vivo analysis, a lower expression of genes encoding tight junction proteins, and a higher expression of genes encoding pro-inflammatory and anti-inflammatory immune activity. In addition, the expression of genes involved in endocrine function, tryptophan metabolism and nutrient transport was modified mostly in the colon. These diet-induced intestinal modifications were associated with changes in the brain tissue expression of genes encoding the blood-brain barrier, endocrine function and short chain fatty acid receptors, mostly in hypothalamic and striatal areas. The integrative approach underlined specific groups of bacteria (Veillonellaceae, Enterobacteriaceae, Lachnospiraceae, Rikenellaceae, and Prevotellaceae) associated with changes in the gut-brain axis. There is a clear influence of the infant diet, even over a short dietary intervention period, on establishment of the microbiota-gut-brain axis.

Gastric digestion of cow milk, almond milk and oat milk in rats.

In this study, gastric digestion of isocaloric and iso-macronutrient cow milk, almond milk and oat milk were compared in rats euthanized at different post-feeding times. The cow milk separated into a curd phase and a liquid phase in the rat stomach. This coagulation of the cow milk led to higher (P < 0.05) protein and lipid retention in the stomach compared with almond milk and oat milk. Almond milk oil bodies aggregated, creamed and rapidly layered in the stomach. This induced a faster (P < 0.05) gastric emptying of proteins (T1/2 = 36 min) compared with cow milk (T1/2 = 89 min) and oat milk (T1/2 = 55 min), and a slower gastric emptying of almond lipids than of almond proteins. In contrast, no significant physical change during the digestion of oat milk was found, with both the proteins and the lipids being steadily emptied from the stomach. This in vivo study provides information on the gastric digestion and emptying (and thereby nutritional characteristics) of plant-based milks compared with animal-based milks, that will be useful for the design of novel plant-based drinks.

The effects of ruminant milk treatments on hippocampal, striatal, and prefrontal cortex gene expression in pigs as a model for the human infant.

While infant formula is usually bovine milk-based, interest in other ruminant milk-based formulas is growing. However, whether different ruminant milk treatments with varying nutrient compositions influence the infant's brain development remains unknown. The aim was to determine the effects of consuming bovine, caprine, or ovine milk on brain gene expression in the early postnatal period using a pig model of the human infant. Starting at postnatal day 7 or 8, pigs were exclusively fed bovine, ovine, or caprine milk for 15 days. The mRNA abundance of 77 genes in the prefrontal cortex, hippocampus, and striatum regions was measured at postnatal day 21 or 22 using NanoString. The expression level of two hippocampal and nine striatal genes was most affected by milk treatments, particularly ovine milk. These modulatory genes are involved in glutamate, gamma-aminobutyric acid, serotonin, adrenaline and neurotrophin signaling and the synaptic vesicle cycle. The expression level of genes involved in gamma-aminobutyric acid signaling was associated with pigs' lactose intake. In contrast, milk treatments did not affect the mRNA abundance of the genes in the prefrontal cortex. This study provides the first evidence of the association of different ruminant milk treatments with brain gene expression related to cognitive function in the first 3 months of postnatal life.

Actinidin reduces gluten-derived immunogenic peptides reaching the small intestine in an in vitro semi-dynamic gastrointestinal tract digestion model.

Actinidin, a cysteine protease in green kiwifruit (Actinidia deliciosa), has been identified as a potential enzyme to hydrolyse gluten within the lumen of the gastrointestinal tract (GIT). The present study aimed to further evaluate the effect of purified actinidin sourced from green kiwifruit on the digestion of gluten and the release of immunogenic peptides during GIT digestion using an in vitro semi-dynamic GIT digestion model. Purified gluten was digested for 180 min with or without actinidin and subsequently analysed for free amino groups (o-phthaldialdehyde) to determine the degree of hydrolysis (DH), gluten R5 epitopes (ELISA), and peptide profiles (mass spectrometry). Strong interactions were observed between treatment (GIT digestion with or without actinidin) and digestion time for the DH of gluten (P < 0.01), amount of free amino groups released into the small intestine (P < 0.01), and amount of gluten epitopes present in the small intestine (P < 0.001). The rate of increase of DH of gluten and the amount of R5 epitopes present in the small intestine during the first 30 min of GIT digestion with actinidin was 0.3%/min and 4.8 ng/g of gluten respectively, whereas it was 0.01%/min and 60.9 ng/g of gluten respectively without actinidin. These results were corroborated by untargeted peptidomics, with a 1.5-fold lower number of known immunogenic epitopes reaching the small intestine at 30 min of GIT digestion when actinidin was present compared to the control. Present results demonstrate that actinidin enhanced the rate of proteolysis of gluten and reduced the number of immunogenic gluten epitopes reaching the small intestine during simulated semi-dynamic GIT digestion.