Sorghum tannin extract impedes in vitro digestibility and fermentability of nutrients in the simulated porcine gastrointestinal tract.

The site and extent of digestion of sorghum nutrients affected by tannins in the intestine are not clarified. Porcine small intestine digestion and large intestine fermentation were simulated in vitro to determine the effects of sorghum tannin extract on the digestion and fermentation characteristics of nutrients in the mimicked porcine gastrointestinal tract. In experiment 1, low-tannin sorghum grain without or with 30 mg/g sorghum tannin extract were digested by porcine pepsin and pancreatin to measure in vitro digestibility of nutrients. In experiment 2, the lyophilized porcine ileal digesta from 3 barrows (Duroc × Landrace × Yorkshire, 27.75 ±â€…1.46 kg) fed the low-tannin sorghum grain without or with 30 mg/g sorghum tannin extract and the undigested residues from experiment 1 were, individually, incubated with fresh pig cecal digesta as inoculums for 48 h to simulate the porcine hindgut fermentation. The results revealed that sorghum tannin extract decreased in vitro digestibility of nutrients both by pepsin hydrolysis or pepsin-pancreatin hydrolysis (P < 0.05). Although enzymatically unhydrolyzed residues provided more energy (P = 0.09) and nitrogen (P < 0.05) as fermentation substrates, the microbial degradation of nutrients from unhydrolyzed residues and porcine ileal digesta were both decreased by sorghum tannin extract (P < 0.05). Regardless of unhydrolyzed residues or ileal digesta as fermentation substrates, microbial metabolites including the accumulative gas production excluding the first 6 h, total short-chain fatty acid and microbial protein content in the fermented solutions were decreased (P < 0.05). The relative abundances of Lachnospiraceae AC2044 and NK4A136 and Ruminococcus_1 was decreased by sorghum tannin extract (P < 0.05). In conclusion, sorghum tannin extract not only directly decreased the chemical enzymatic digestion of nutrients in the simulated anterior intestine, but also directly inhibited the microbial fermentation including microbial diversities and metabolites in the simulated posterior intestine of pigs. The experiment implies that the decreased abundances of Lachnospiraceae and Ruminococcaceae by tannins in the hindgut may weaken the fermentation capacity of microflora and thus impair the nutrient digestion in the hindgut, and ultimately reduce the total tract digestibility of nutrients in pigs fed high tannin sorghum.

Sorghum tannins may be potent negative factor to impede the nutrient utilization in sorghum grain fed to pigs. Unfortunately, the site, rate and extent of digestion of sorghum nutrients affected by tannins in the intestine remain unclear. Therefore, this experiment was conducted to determine the effects of sorghum tannin extract on the digestion and fermentation characteristics of nutrients in the in vitro simulated porcine gastrointestinal tract. The results revealed that sorghum tannin extract decreased the chemical enzymatic digestion of nutrients in the simulated anterior intestine, and inhibited the hindgut microbial fermentation including microbial diversities and metabolites (accumulative gas and short-chain fatty acid production) in the simulated posterior intestine of pigs. The experiment implies that the decreased abundances of Lachnospiraceae and Ruminococcaceae by tannins in the hindgut may weaken the fermentation capacity of microflora and thus impair the nutrient digestion in the hindgut, and ultimately may reduce the total tract nutrient digestibility in pigs fed high tannin sorghum.

Effects of recombinant human gastric lipase and pancreatin during in vitro pediatric gastro-intestinal digestion.

The aim of the study was to implement a gastric digestion step using recombinant human gastric lipase (rHGL) in an in vitro pediatric gastro-intestinal digestion model to achieve a physiologically relevant gastric contribution to total gastro-intestinal lipid digestion. A commercial infant formula (NAN Comfort stage 1 (NAN1)) with 3.4% lipid and an in-lab prepared oil-in-water emulsion, emulsified with soy phosphatidylcholine (SPCemul), with 3.5% lipid (oil-blend containing Akonino NS, MEG-3 and ARASCO oils) were subjected to in vitro gastro-intestinal digestion. To achieve a physiologically relevant level of gastric digestion, 50 min of in vitro gastric digestion, using either 0, 3.75 or 7.5 TBU mL-1 rHGL, was followed by 90 min of in vitro intestinal digestion, using either 0 or 26.5 TBU mL-1 pancreatic triglyceride lipase (PTL) from porcine pancreatin. The digestion of the substrates was assessed using titration-based quantification supported by HPLC-ELSD analysis. In vitro gastric digestion of NAN1 and SPCemul with either 3.75 or 7.5 TBU mL-1 rHGL contributed with 10-27% of the total gastro-intestinal digestion, corresponding to the reported contribution in human infants. At the end of the gastro-intestinal digestion (t = 140 min), the combined lipolytic effect of rHGL and PTL was additive during digestion of SPCemul, but not for the digestion of NAN1, as all lipase activity combinations resulted in a similar degree of NAN1 digestion. The effect of gastric digestion with rHGL on total digestion therefore appeared to be substrate dependent. To conclude, a gastric digestion step using rHGL resulting in physiologically relevant gastric contribution to the observed gastro-intestinal digestion was successfully implemented into an in vitro pediatric gastro-intestinal digestion model.

Antibacterial, anti-biofilm activity and mechanism of action of pancreatin doped zinc oxide nanoparticles against methicillin resistant Staphylococcus aureus.

Staphylococcus aureus are known to cause diseases from normal skin wound to life intimidating infections. Among the drug resistant strain, management of methicillin resistant Staphylococcus aureus (MRSA) is very difficult by using conventional antibiotic treatment. Both Zinc oxide nanoparticles (ZnONPs) and pancreatin (PK) are known to have antibacterial activity. Our main objective is to dope PK on ZnONPs to reduced zinc-oxide toxicity but increased anti-bacterial and anti-biofilms activity. In present study, we showed that, functions of zinc oxide nanoparticles with pancreatin enzyme (ZnONPs-PK) have anti-bacterial, anti-biofilms, anti-motility and anti-virulence properties against MRSA. Moreover, ZnONPs-PK were more potent to eradicate MRSA than only ZnONPs and PK. Application of the produced nano-composites as treatment on infected swine dermis predominantly reflects the potential treatment property of it. The vancomycin sensitivity of MRSA was significantly increased on application with ZnONPs-PK. Further study revealed cell membrane was the target of the ZnONPs-PK and that leads to oxidative damage of the cells. The produced nanoparticles were found completely non-toxic to human's keratinocytes and lung epithelial cell lines at its bactericidal concentration. Overall, this study emphasizes the potential mechanisms underlying the selective bactericidal properties of ZnONPs-PK against MRSA. This novel nanoparticle strategy may provide the ideal solution for comprehensive management of MRSA and its associated diseases with minimising the use of antibiotics.

Short note: Infliximab recovery in a simulated intestinal fluid of the upper intestine tract.

BACKGROUND: The oral administration of Infliximab (IFX) antibody would ensure a direct action on inflamed intestinal tissues without side effects. Thus, investigations about its resilience within the intestinal environment are required. OBJECTIVE: Quantify the IFX recovery in a simulated upper intestinal environment. METHODS: IFX was incubated for different times until 120 min in simulated intestinal fluid (SIF) which differed (i) for pH (7.2 vs 6.8, Exp 1), (ii) for addition or not with pancreatin (Exp 2) and (iii) for addition or not with bovine serum albumin in presence of pancreatin (BSA, Exp 3). RESULTS: In Exp 1 the IFX incubated without pancreatin was degraded by about 15% by SIF pH change from 7.2 to 6.8 and after 120 min it was reduced by about 20%. In Exp 2 the presence of pancreatin determined an intense and rapid IFX degradation (recovery < 33%, within 30 min), but when BSA was added to simulate the presence of food protein (Exp 3) the IFX half-life ranged between 59 and 70 min. CONCLUSIONS: A discrete in vitro stability of IFX in the upper intestine environment was demonstrated, if food protein is available and competes with pancreatin proteases.

Development of an Animal Model for Inducing Various Degrees of Severity of Incontinence-Associated Dermatitis.

PURPOSE: The purpose of this study was to describe the biological changes after incontinence-associated dermatitis (IAD) induction by pancreatin in the guinea pigs and to explore the potentially appropriate timing and pancreatin concentration for IAD induction with different severity. DESIGN: In vivo, experimental study. SUBJECTS AND SETTING: An experimental animal model (guinea pig) in a controlled laboratory setting was used for investigation. METHODS: We developed an IAD model in guinea pigs by occluded application of 1%, 5%, and 10% pancreatin solutions for 1, 3, and 5 days, respectively. The irritant was applied to the posterior aspect of shaved guinea pigs. We used an adapted visual scoring system to evaluate IAD and its severity. We also measured differences of the fluid absorption rate as a proxy for transepidermal water loss and enzyme-linked immunosorbent assays of interleukin 2 and interferon-γ expression as indicators of IAD-related inflammation. Analysis of variance (ANOVA) was used to examine group differences. RESULTS: Higher pancreatin concentrations led to more severe skin responses and higher mean visual scale scores, yet the statistically score differences were only observed in the 1% and 5% pancreatin groups after 3 and 5 days of exposure compared with 1 day of exposure (P < .05). The average absorbed fluid rate increased from 1 to 3 days of exposure and reached a plateau at 3 days; significant differences were observed in 3 and 5 days of exposure (P < .05) when compared with 1 day of exposure but not between 3 and 5 days of exposure. CONCLUSIONS: Exposure of a guinea pig animal model to 1%, 5%, and 10% pancreatin solutions over a 3-day period induced IAD with different levels of severity. Additional studies using this model are warranted.

Interaction between Various Apple Procyanidin and Staphylococcal Enterotoxin A and Their Inhibitory Effects on Toxin Activity.

In this study, we investigated the interaction between apple polyphenols (AP; mainly consisting of procyanidin (PC) from an apple) and staphylococcal enterotoxin A (SEA), and the inhibitory effects of AP on SEA activity. According to the degree of polymerization, in particularly highly polymerized PC (more than pentamer) strongly interacted with SEA. The binding affinity of AP with SEA molecules was determined using Biacore analysis. AP reacted with SEA immobilized on a Biacore sensor chip. After treatment with pepsin and pancreatin, to examine the changes of binding affinity of AP in intragastric conditions, AP maintained interaction with SEA. We examined whether AP inhibits the proliferation and interferon-γ (IFN-γ) production induced by SEA in mouse spleen cells. AP strongly inactivated the proliferation and IFN-γ production induced by SEA. These results suggest that AP, which has a higher degree of polymerization, inactivates stronger biological activity of SEA through interaction with SEA. Our studies are the first to demonstrate the relationship between the degree of polymerization of AP and the inhibitory effects on SEA activities.

Incorporation of in vitro digestive enzymes in an intestinal epithelial cell line model for protein hazard identification.

Relatively few proteins in nature produce adverse effects following oral exposure. Of those that do, effects are often observed in the gut, particularly on intestinal epithelial cells (IEC). Previous studies reported that addition of protein toxins to IEC lines disrupted monolayer integrity but innocuous dietary proteins did not. Studies presented here investigated the effects of innocuous (bovine serum albumin, ß-lactoglobulin, RuBisCO, fibronectin) or hazardous (phytohaemagglutinin-E, concanavalin A, wheat germ agglutinin, melittin) proteins that either were untreated or exposed to digestive enzymes prior to addition to Caco-2 human IEC line monolayers. At high concentrations intact fibronectin caused an increase in monolayer permeability but other innocuous proteins did not whether exposed to digestive enzymes or not. In contrast, all untreated hazardous proteins and those that were resistant to digestion (ex. wheat germ agglutinin) disrupted monolayer integrity. However, proteins sensitive to degradation by digestive enzymes (ex. melittin) did not adversely affect monolayers when exposed to these enzymes prior to addition to IEC line monolayers. These results indicate that in vitro exposure of proteins to digestive enzymes can assist in differentiating between innocuous and hazardous proteins as another component to consider in the overall weight of evidence approach in protein hazard assessment.

Effects of the Food Manufacturing Chain on the Viability and Functionality of Bifidobacterium animalis through Simulated Gastrointestinal Conditions.

The viability and functionality of probiotics may be influenced by industrial production processes resulting in a decrease in probiotic efficiency that benefit the health of humans. This study aimed to investigate the probiotic characteristics of Bifidobacterium strains isolated from fecal samples of healthy Thai infants. In the present work, three local strains (BF014, BF052, and BH053) belonging to Bifidobacterium animalis showed a great resistance against conditions simulating the gastrointestinal tract. Among these, B. animalis BF052 possessed considerable probiotic properties, including high acid and bile tolerance, strong adhesion capability to Caco-2 cells, and inhibitory activity against pathogens including Salmonella typhimurium and Vibrio cholerae. This strain also exhibited a high survival rate compared to commercial strains during storage in a wide variety of products, including pasteurized milk, soy milk, drinking yogurt, and orange juice. The impact of food processing processes as well as the freeze-drying process, storage of freeze-dried powders, and incorporation of freeze-dried cells in food matrix on probiotic properties was also determined. The stability of the probiotic properties of the BF052 strain was not affected by food processing chain, especially its resistance in the simulated gastrointestinal conditions and its adherence ability to Caco-2 cells. It indicates that it satisfies the criteria as a potential probiotic and may be used as an effective probiotic starter in food applications.

Yarrowia lipolytica Lipase 2 Is Stable and Highly Active in Test Meals and Increases Fat Absorption in an Animal Model of Pancreatic Exocrine Insufficiency.

BACKGROUND & AIMS: Pancreatic exocrine insufficiency (PEI) reduces pancreatic secretion of digestive enzymes, including lipases. Oral pancreatic enzyme replacement therapy (PERT) with pancreatin produces unsatisfactory results. The lipase 2 produced by the yeast Yarrowia lipolytica (YLLIP2; GenBank: AJ012632) might be used in PERT. We investigated its ability to digest triglycerides in a test meal and its efficacy in reducing fecal fat in an animal model of PEI. METHODS: YLLIP2 was produced by genetically engineered Y lipolytica and purified from culture media. YLLIP2 or other gastric (LIPF) and pancreatic (PNLIPD) lipases were added to a meal paste containing dietary triglycerides, at a range of pH values (pH 2-7), with and without pepsin or human bile and incubated at 37°C. We collected samples at various time points and measured lipase activities and stabilities. To create an animal model of PEI, steatorrhea was induced by embolization of the exocrine pancreas gland and pancreatic duct ligation in minipigs. The animals were given YLLIP2 (1, 4, 8, 40, or 80 mg/d) or pancreatin (100,000 US Pharmacopeia lipase units/d, controls) for 9 days. We then collected stool samples, measured fat levels, and calculated coefficient of fat absorption (CFA) values. RESULTS: YLLIP2 was highly stable and poorly degraded by pepsin, and had the highest activity of all lipases tested on meal triglyceride at pH 4-7 (pH 6 with bile: 94 ± 34 U/mg; pH 4 without bile: 43 ± 13 U/mg). Only gastric lipase was active and stable at pH 3, whereas YLLIP2 was sensitive to pepsin hydrolysis after pH inactivation. From in vitro test meal experiments, the lipase activity of YLLIP2 (10 mg) was estimated to be equivalent to that of pancreatin (1200 mg; 100,000 US Pharmacopeia units) at pH 6. In PEI minipigs, CFA values increased from 60.1% ± 9.3% before surgery to 90.5% ± 3.2% after administration of 1200 mg pancreatin (P < .05); CFA values increased to a range of 84.6% ± 3.0% to 90.0% ± 3.8% after administration of 4-80 mg YLLIP2 (P < .05). CONCLUSIONS: The yeast lipase YLLIP2 is stable and has high levels of activity against test meal triglycerides in a large pH range, with and without bile. Oral administration of milligram amounts of YLLIP2 significantly increased CFA values, similar to that of 1.2 g pancreatin, in a minipig model of PEI.

Nitrogen sources impact hydrogen production by Escherichia coli using cheese whey as substrate.

The impact of nitrogen source on hydrogen production by Escherichia coli WDHL (ΔhycAΔlacI) strain using cheese whey as a substrate was evaluated. To improve the assimilation of complex proteins such as lactalbumin, we assessed treatment with a protease. Also, five external nitrogen sources were tested: NH4Cl, (NH4)2SO4, urea, yeast extract, and tryptone. The treatments in 120 mL serological bottles with pancreatin 1,000 mg/L produced 1.75-fold more hydrogen than the cultures without pancreatin. In the bottle cultures supplemented with yeast extract or tryptone 5 g/L, hydrogen production increased up to 3.2- and 3.5-fold, respectively, whereas inorganic salts and urea had no statistical difference with respect to the control cultures. In 1-L bioreactors, the use of tryptone improved 2.1-fold hydrogen production. Tryptone or yeast extract enable the total consumption of lactose in 40 h, whereas in the control assay the lactose was not completely consumed. Our results demonstrate that it is necessary to select an adequate nitrogen source, which allows both carbon source consumption and high hydrogen production.

Effects of porcine pancreatic enzymes on the pancreas of hamsters. Part 1: basic studies.

CONTEXT: Porcine pancreatic enzymes (PPE) extracted from glandular stomach has been used for the treatment of pancreatic cancer patients. Unfortunately, no information is available on the in vitro and in vivo effect on the pancreas and other tissues. OBJECTIVE: We used Syrian Golden hamsters, a unique pancreatic cancer model, to obtain basic information on PPE for its eventual use for the treatment of pancreatic cancer. DESIGN: PPE was used in different concentrations in vitro and in vivo. The stability of the enzyme in the water solution was investigated. It was given to the hamsters by gavage in concentrations of 1g/kg and 400 mg/kg for short periods and in aqueous solution for 65 days. Plasma enzyme and insulin, the size of islets and the number of the insulin cells per islet were examined. RESULTS: The enzyme activity of PPE was maintained in water solution for at least 24 hours. Due to its content of calcium chloride it showed a high toxicity to normal and malignant hamster pancreatic cancer cells and human pancreatic cancer cell lines in vitro. PPE did not alter the plasma pancreatic enzyme levels regardless of the dose, duration and application route. On the contrary, PPE reduced their levels significantly. Remarkably, it also reduced the level of insulin, the size of the islets and the number of insulin cells in the islets significantly. CONCLUSION: The results imply that PPE does not enter the blood circulation but it appears to slow down the function of both the exocrine and endocrine pancreas.

Effects of porcine pancreatic enzymes on the pancreas of hamsters. Part 2: carcinogenesis studies.

CONTEXT: Our previous study suggested that porcine pancreatic extract in hamsters with peripheral insulin resistance, normalizes insulin output, islet size and pancreatic DNA synthetic rate. It also inhibited the growth of human pancreatic cancer cells in nude mice. OBJECTIVE: To examine the potential value of the porcine pancreatic extract in controlling pancreatic carcinogenesis in this model, the present experiment was performed. DESIGN: Hamsters were fed a high fat diet and four weeks later when insulin resistance emerges, they were divided into two groups. One group received 1 g/kg BW of porcine pancreatic extract in drinking water and the other group received tap water. One week later, when insulin output normalizes in porcine pancreatic extract-treated hamsters, a single subcutaneous injection of N-nitrosobis-(2-oxopropyl) amine (BOP) at a dose of 40 mg/kg BW was given to all hamsters. The experiment was terminated at 43 weeks after the porcine pancreatic extract treatment. The number and size of pancreatic tumors, blood glucose, insulin, amylase and lipase levels, the average size of islets and the number of insulin cells/islets were determined. RESULTS: The incidence of pancreatic cancer was significantly lower in the porcine pancreatic extract group (P=0.043), as well as the plasma insulin level and the size of the islets in the porcine pancreatic extract group were significantly lower (P<0.001) than in the control group. No significantly differences were found in the glucose level between the groups. CONCLUSION: These results show that porcine pancreatic extract has a potential to inhibit pancreatic cancer growth.

Ileal digestibility of amino acids, phosphorus, phytate and energy in pigs fed sorghum-based diets supplemented with phytase and Pancreatin®.

The effects of phytase supplementation on the apparent ileal digestibility (AID) of amino acids (AA) have been inconsistent. Two experiments evaluated the effect of providing a mixture of pancreatic enzymes (Pancreatin(®) ) to growing pigs fed sorghum-soybean meal diets supplemented with phytase on the AID of AA, energy, and phosphorus (P), as well as the ileal digestibility (ID) of phytate; there were four periods per experiment. In Experiment 1, eight pigs (BW 22.1±1.3 kg) were fitted with a T-cannula at the distal ileum. Each period consisted of 9 days; 7 days for diet adaptation, and 2 days for digesta collection. Treatments (T) were: (i) basal sorghum-soybean meal diet, (ii) basal diet plus Pancreatin®, (iii) basal diet plus phytase and (iv) basal diet plus phytase and Pancreatin®. Phytase increased the digestibilities of phytate and P (p<0.001), but did not affect the AID of AA and energy (p>0.10). Except for methionine (p=0.07), Pancreatin® did not affect the AID of AA. Phytase and Pancreatin® did not interact (p>0.10). Experiment 2 was similar to Experiment 1, but Pancreatin® was infused into duodenum. Pancreatin® infusion did not affect the AID of AA (p>0.10); and tended to reduce (p=0.09) the AID of lysine. Phytase × Pancreatin® interactions were not observed (p>0.10). In conclusion, phytase and Pancreatin® did not improve the AID of AA in growing pigs fed sorghum-soybean meal diets indicating that phytates did not affect AA digestibility.

Fragment analogs as better mimics of obestatin.

Obestatin is a twenty three amino acid peptide produced in the stomach by post translational modification of the preproghrelin gene. Since its discovery in 2005, many studies have shown that obestatin reduces feed intake and gain in body weight in rodents. Studies from our laboratory have shown the N-terminal thirteen residues mimic obestatin the best and residues 6-18 reduce epididymal fat significantly in adult male mice. In this study we have tried to increase the efficacy of these fragments. As an initial step, we have substituted G(8) with alpha-aminoisobutyricacid(Aib,U) and F(5) with cyclohexylalanine(Cha) in the N-terminal peptide to obtain two modified peptides and modified the middle fragment (residues 6-18) by substituting both the glycine residues at position 3 and 8 with alpha-aminoisobutyricacid(U). The rationale being, unusual amino acids could protect the peptides from immediate degradation and Aib would also induce secondary structure in these unstructured peptides. The N-terminal fragment with the G(8)U substitution fared the best. It reduced food intake, gain in body weight, levels of cholesterol and triglycerides in the blood, epididymal and perirenal fat in adult male mice similar to that of obestatin. The middle fragment with G(3,8)U double substitution was the second best.

Molecular and probiotic characterization of bacteriocin-producing Enterococcus faecium strains isolated from nonfermented animal foods.

AIMS: The characterization of four novel bacteriocin-producing enterococcal strains, isolated from nonfermented animal foods, was carried out with a view to evaluate their potential application as probiotics in raw and processed foodstuffs. METHODS AND RESULTS: 16S rRNA sequencing and random amplification of polymorphic DNA-polymerase chain reaction (RAPD-PCR) analysis allowed the identification and intra-specific grouping of Enterococcus faecium strains, which inhibited the growth of four relevant food-borne pathogenic and spoilage species. Enterococcus faecium strains exhibited remarkable probiotic profiles, being able to survive to pH 3.0 and to the presence of bile salts, pancreatin and pepsin. Enterococcus faecium strains evaluated did not exhibit bile salt hydrolase or haemolytic activity, but showed good adhesion properties, also exhibiting sensitivity to clinically relevant antimicrobial agents. CONCLUSIONS: In our study, DNA sequencing of the 16S rRNA gene and RAPD-PCR analysis were equally discriminatory for typing E. faecium strains. This study also confirmed the potential tolerance and survival of E. faecium strains isolated from nonfermented animal foods to the gastrointestinal tract. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents the first report on potential probiotic E. faecium strains isolated from nonfermented meat and fish. Their moderate heat resistance opens the way to their potential use as probiotics in minimally processed foods subjected to moderate heat processing.

Assessing potential effects of inulin and probiotic bacteria on Fe availability from common beans (Phaseolus vulgaris L.) to Caco-2 cells.

Inulin, a prebiotic, may enhance intestinal Fe absorption. Our objective was to assess the effects of supplemental inulin and 2 probiotic bacteria (B. infantis and L. acidophillus) on Fe availability to Caco-2 cells from common white and red beans (Phaseolus vulgaris L.). Cooked beans were mixed or not with supplemental inulin (4%, w/w), and then subjected to simulated gastrointestinal digestion (pepsin, pH 2; pancreatin, pH 7.2). Subsequently, the digests were incubated overnight with and without B. infantis or L. acidophilus. Ferritin formation in Caco-2 cells was used to evaluate Fe uptake. Total soluble phenols (Folin-Ciocalteau) and phytate (HPLC-electrochemical detection) were quantified, and the flavonoids profile (HPLC-PDA/UV detection) was monitored in the digests. Supplemental inulin did not affect Fe uptake from white nor red beans. Incubation with B. infantis increased total soluble phenols (TSP) in the digests and decreased Fe uptake. Incubation with L. acidophilus decreased TSP in the digest and increased Fe uptake. Variations in Fe uptake were not associated with soluble phytate concentrations in the digests. The largest change in flavonoids profile were found in the digests incubated with L. acidophilus, which decreased the soluble concentration of astragalin (kaempferol-3-O-glucoside). These results suggest that certain probiotics could increase Fe uptake from common beans.

Stability of anthocyanins from commercial black currant juice under simulated gastrointestinal digestion.

Anthocyanins are effective antioxidants but they have also been proposed to have other biological activities independent of their antioxidant capacities that produce health benefits. Examples range from inhibition of cancer cell growth in vitro, induction of insulin production in isolated pancreatic cells, reduction of starch digestion through inhibition of a-glucosidase activity, suppression of inflammatory responses as well as protection against age-related declines in cognitive behavior and neuronal dysfunction in the central nervous system. However, to achieve any biological effect in a specific tissue or organ, anthocyanins must be bioavailable; i.e. effectively absorbed from the gastrointestinal tract (GIT) into the circulation and delivered to the appropriate location within the body. In this study, we assess the stability of anthocyanins from commercial Black currant (Ribes nigrum L.) juice using an in vitro digestion procedure that mimics the physiochemical and biochemical conditions encountered in the gastrointestinal tract (GIT). The main objective of this work was the evaluation of stability of anthocyanins during in vitro digestion in gastric and intestinal fluid regarding whether appropriate enzyme (pepsin or pancreatin) was added or not. Anthocyanins present in commercial black currant juice remain stable during in vitro digestion in gastric fluid regardless whether pepsin was added into the medium or not. Also, they remain stable during in vitro digestion in simulated intestinal fluid without pancreatin. The stability studies of anthocyanins in the intestinal fluid containing pancreatin indicated reduced stability, which also mainly contribute to slight reduction of total anthocyanins content (-1.83%) in commercial black currant juice.

Redefining the role of ectoderm in somitogenesis: a player in the formation of the fibronectin matrix of presomitic mesoderm.

The absence of ectoderm impairs somite formation in cultured presomitic mesoderm (PSM) explants, suggesting that an ectoderm-derived signal is essential for somitogenesis. Here we show in chick that the standard enzymatic treatments used for explant isolation destroy the fibronectin matrix surrounding the anterior PSM, which fails to form somites when cultured for 6 hours. By contrast, explants isolated with collagenase retain their fibronectin matrix and form somites under identical culture conditions. The additional presence of ectoderm enhances somite formation, whereas endoderm has no effect. Furthermore, we show that pancreatin-isolated PSM explants cultured in fibronectin-supplemented medium, form significantly more somites than control explants. Interestingly, ectoderm is the major producer of fibronectin (Fn1) transcripts, whereas all but the anterior-most region of the PSM expresses the fibronectin assembly receptor, integrin alpha5 (Itga5). We thus propose that the ectoderm-derived fibronectin is assembled by mesodermal alpha5beta1 integrin on the surface of the PSM. Finally, we demonstrate that inhibition of fibronectin fibrillogenesis in explants with ectoderm abrogates somitogenesis. We conclude that a fibronectin matrix is essential for morphological somite formation and that a major, previously unrecognised role of ectoderm in somitogenesis is the synthesis of fibronectin.

In vitro assessment of the gastrointestinal transit tolerance of taxonomic reference strains from human origin and probiotic product isolates of Bifidobacterium.

Next to health promoting effects, the functional aspect of probiotic strains also involves their capacity to reach the colon as viable metabolically active cells. The present study aimed to assess the potential of 24 probiotic product isolates and 42 human reference strains of Bifidobacterium to survive gastrointestinal transit under in vitro conditions. The survival capacity of exponential and stationary phase cultures upon exposure to gastric and small intestinal juices was determined using a recently developed microplate-based assay in combination with the LIVE/DEAD BacLight Bacterial Viability kit. All 66 strains tested displayed a considerable loss in viability during exposure to an acidic pepsin containing solution (pH 2.0). Among the 10 taxa tested, cultures of B. animalis ssp. lactis appeared to be most capable to survive gastric transit. Although to a lesser extent, the presence of bile salts also affected the viability of most of the strains tested. Except for 3 strains, all 66 strains showed bile salt hydrolase activity using an agar-based assay. In contrast, the bifidobacterial strains used in this study appeared to possess a natural ability to survive the presence of pancreatin (pH 8.0). Although the effect was not significant, a slightly enhanced tolerance to gastrointestinal transit was observed when cells were in the stationary phase, especially when exposed to acid, compared with cells being in the exponential phase. Survival in the gastrointestinal tract appeared to be largely strain-dependent and hence implies that different strains will likely display a different behavior in functionality. The assay used in this study allows an initial assessment of strains for use as probiotic cultures prior to selecting potential candidate strains for further investigation in vivo.

Expression of the mucus adhesion genes Mub and MapA, adhesion-like factor EF-Tu and bacteriocin gene plaA of Lactobacillus plantarum 423, monitored with real-time PCR.

Expression of the mucus adhesion genes Mub and MapA, adhesion-like factor EF-Tu and bacteriocin gene plaA by Lactobacillus plantarum 423, grown in the presence of bile, pancreatin and at low pH, was studied by real-time PCR. Mub, MapA and EF-Tu were up-regulated in the presence of mucus, proportional to increasing concentrations. Expression of MapA was up-regulated in the presence of 3.0 g/l bile and 3.0 g/l pancreatin at pH 6.5. Similar results were recorded in the presence of 10.0 g/l bile and 10.0 g/l pancreatin at pH 6.5. Expression of Mub was down-regulated in the presence of bile and pancreatin, whilst the expression of EF-Tu and plaA remained unchanged. Expression of Mub and MapA remained unchanged at pH 4.0, whilst expression of EF-Tu and plaA were up-regulated. Expression of MapA was down-regulated in the presence of 1.0 g/l l-cysteine HCl, suggesting that the gene is regulated by transcription attenuation that involves cysteine.