Dietary Isomalto/Malto-Polysaccharides Increase Fecal Bulk and Microbial Fermentation in Mice

Scope: The prevalence of metabolic-syndrome-related disease has strongly increased. Nutritional intervention strategies appear attractive, particularly with novel prebiotics. Isomalto/malto-polysaccharides (IMMPs) represent promising novel prebiotics that promote proliferation of beneficial bacteria in vitro. The present study investigates for the first time the in vivo effects of IMMP in mice. Methods and results: C57BL/6 wild-type mice received control or IMMP-containing (10%, w/w) diets for 3 weeks. IMMP leads to significantly more fecal bulk (+26%, p < 0.05), higher plasma non-esterified fatty acids (colorimetric assay, +10%, p < 0.05), and lower fecal dihydrocholesterol excretion (mass spectrometry, −50%, p < 0.05). Plasma and hepatic lipid levels (colorimetric assays following lipid extraction) are not influenced by dietary IMMP, as are other parameters of sterol metabolism, including bile acids (gas chromatography/mass spectrometry). IMMP is mainly fermented in the cecum and large intestine (high-performance anion exchange chromatography). Next-generation sequencing demonstrates higher relative abundance of Bacteroides and butyrate producers (Lachnospiraceae, Roseburia Odoribacter) in the IMMP group. Conclusion: The combined results demonstrate that IMMP administration to mice increases fecal bulk and induces potentially beneficial changes in the intestinal microbiota. Further studies are required in disease models to substantiate potential health benefits.</p

The association between breastmilk oligosaccharides and faecal microbiota in healthy breastfed infants at two, six, and twelve weeks of age

Several factors affect gut microbiota development in early life, among which breastfeeding plays a key role. We followed 24 mother-infant pairs to investigate the associations between concentrations of selected human milk oligosaccharides (HMOs) in breastmilk, infant faeces, and the faecal microbiota composition in healthy, breastfed infants at two, six and 12 weeks of age. Lactation duration had a significant effect on breastmilk HMO content, which decreased with time, except for 3-fucosyllactose (3FL) and Lacto-N-fucopentaose III (LNFP III). We confirmed that microbiota composition was strongly influenced by infant age and was associated with mode of delivery and breastmilk LNFP III concentration at two weeks, with infant sex, delivery mode, and concentrations of 3′sialyllactose (3′SL) in milk at six weeks, and infant sex and Lacto-N-hexaose (LNH) in milk at 12 weeks of age. Correlations between levels of individual breastmilk HMOs and relative abundance of OTUs found in infant faeces, including the most predominant Bifidobacterium OTUs, were weak and varied with age. The faecal concentration of HMOs decreased with age and were strongly and negatively correlated with relative abundance of OTUs within genera Bifidobacterium, Parabacteroides, Escherichia-Shigella, Bacteroides, Actinomyces, Veillonella, Lachnospiraceae Incertae Sedis, and Erysipelotrichaceae Incertae Sedis, indicating the likely importance of these taxa for HMO metabolism in vivo.</p

Effect of the prebiotic fiber inulin on cholesterol metabolism in wildtype mice

Dietary non-digestible carbohydrates are perceived to improve health via gut microbiota-dependent generation of products such as short-chain fatty acids (SCFA). In addition, SCFA are also precursors for lipid and cholesterol synthesis potentially resulting in unwanted effects on lipid metabolism. Inulin is a widely used model prebiotic dietary fiber. Inconsistent reports on the effects of inulin on cholesterol homeostasis have emerged in humans and preclinical models. To clarify this issue, the present study aimed to provide an in-depth characterization of the effects of short-chain (sc)- and long-chain (lc)- inulin on cholesterol synthesis, absorption and elimination in mice. Feeding wildtype C57BL/6J mice diets supplemented with 10% (w/w) of either sc- or lc-inulin for two weeks resulted in approximately 2.5-fold higher fecal SCFA levels (P < 0.01) compared with controls, but had no significant effects on plasma and liver lipids. Subtle shifts in fecal and plasma bile acid species were detected with beta-muricholic acid increasing significantly in plasma of the inulin fed groups (1.7-fold, P < 0.05). However, neither sc-inulin nor lc-inulin affected intestinal cholesterol absorption, mass fecal cholesterol excretion or trans-intestinal cholesterol excretion (TICE). Combined, our data demonstrate that sc- and lc-inulin have no adverse effects on cholesterol metabolism in mice despite increased generation of SCFA.</p

Combining HPAEC-PAD, PGC-LC-MS, and 1D ¹H NMR to Investigate Metabolic Fates of Human Milk Oligosaccharides in 1-Month-Old Infants:A Pilot Study

A solid-phase extraction procedure was optimized to extract 3-fucosyllactose and other human milk oligosaccharides (HMOs) from human milk samples separately, followed by absolute quantitation using high-performance anion-exchange chromatography-pulsed amperometric detection and porous graphitized carbon-liquid chromatography-mass spectrometry, respectively. The approach developed was applied on a pilot sample set of 20 human milk samples and paired infant feces collected at around 1 month postpartum. One-dimensional 1H nuclear magnetic resonance spectroscopy was employed on the same samples to determine the relative levels of fucosylated epitopes and sialylated (Neu5Ac) structural elements. Based on different HMO consumption patterns in the gastrointestinal tract, the infants were assigned to three clusters as follows: complete consumption; specific consumption of non-fucosylated HMOs; and, considerable levels of HMOs still present with consumption showing no specific preference. The consumption of HMOs by infant microbiota also showed structure specificity, with HMO core structures and Neu5Ac(α2-3)-decorated HMOs being most prone to degradation. The degree and position of fucosylation impacted HMO metabolization differently. </p

Variation in gastrointestinal metabolization of prebiotics : Learnings from human milk oligosaccharides and modified starch

Prebiotics and its close interactions with human gut microbiota is gaining attention in recent years, as it exerts multiple impacts on human health. Studying the metabolization of metabolic fates of indigestible oligo- and poly- saccharides in the gastro-intestinal tract is vital for understanding the mechanism of how the carbohydrates influence gut environment and immune system. The current thesis describes the metabolization of indigestible carbohydrates, human milk oligosaccharides (HMOs) and isomalto/malto-polysaccharides (IMMPs) by infant or adult gut microbiota using in vivo or in vitro study approaches.HMOs are considered as the first natural prebiotic for life, providing essential benefits to support infant health. Concentrations of 19 major HMOs present in human milk and infant fecal samples were obtained by combining high performance anion exchange chromatography - pulsed amperometric detection (HPAEC-PAD) ans porous graphitized carbon - liquid chromatography mass spectrometry (PGC-LC-MS). Relative levels of fucosylated and sialylated HMO structural elements was obtained by one-dimensional 1H nuclear magnetic resonance (1D 1H NMR). &nbsp;Mother milk and paired infant fecal samples at one-month postpartum from the KOALA cohort study and at two-, six-, and 12- weeks postpartum from the BINGO cohort study were analyzed for HMOs present. Fecal microbiota composition was characterized using Illumina HiSeq amplicon 16S rRNA sequencing. The results showed inter-individual variations regarding HMO synthesis by mothers and metabolism by infant gut microbiota during the first three months of life. HMOs with different structural elements were differentially consumed when passing through infant gut, and their metabolic fates depended on both linkage types and decoration of fucose/sialic acid moieties. Three distinct consumption patterns of HMOs were found among infants under three months of age, which correlated to three microbial community clusters found in the infant fecal samples. Caesarean section and early exposure to hospital/clinic associated surroundings were found to delay the gradual progression of infant gut development stages. Several phylotypes (OTUs) within specific genera were suggested to be the key taxa responding for HMO metabolization.&nbsp;Prebiotic fermentation behaviors of IMMPs were studied using a batchwise in vitro fermentation model with human fecal inoculum. Structural degradation on molecular levels, metabolites production, enzyme expression and microbiota composition changes during in vitro fermentation were linked together, pointed out a slow-fermenting behavior and structure-specific prebiotic effects of IMMPs.Overall, these findings provide valuable evidence for researchers and industry to identify functional structures of prebiotics for humans at different developmental stages.&nbsp;&nbsp;&nbsp

The Potential of Pectins to Modulate the Human Gut Microbiota Evaluated by In Vitro Fermentation: A Systematic Review

Pectin is a dietary fiber, and its health effects have been described extensively. Although there are limited clinical studies, there is a growing body of evidence from in vitro studies investigating the effect of pectin on human gut microbiota. This comprehensive review summarizes the findings of gut microbiota modulation in vitro as assessed by 16S rRNA gene-based technologies and elucidates the potential structure-activity relationships. Generally, pectic substrates are slowly but completely fermented, with a greater production of acetate compared with other fibers. Their fermentation, either directly or by cross-feeding interactions, results in the increased abundances of gut bacterial communities such as the family of Ruminococcaceae, the Bacteroides and Lachnospira genera, and species such as Lachnospira eligens and Faecalibacterium prausnitzii, where the specific stimulation of Lachnospira and L. eligens is unique to pectic substrates. Furthermore, the degree of methyl esterification, the homogalacturonan-to-rhamnogalacturonan ratio, and the molecular weight are the most influential structural factors on the gut microbiota. The latter particularly influences the growth of Bifidobacterium spp. The prebiotic potential of pectin targeting specific gut bacteria beneficial for human health and well-being still needs to be confirmed in humans, including the relationship between its structural features and activity

Response Surface Optimization of Extraction Conditions for the Active Components with High Acetylcholinesterase Inhibitory Activity and Identification of Key Metabolites from <i>Acer truncatum</i> Seed Oil Residue

The State Council of China has called for the comprehensive development and utilization of Acer truncatum resources. However, research on one of its by-products, namely seed oil residue (ASR), from seed oil extraction is seriously insufficient, resulting in a waste of these precious resources. We aimed to optimize the conditions of ultrasound-assisted extraction (UAE) using a response surface methodology to obtain high acetylcholinesterase (AChE) inhibitory components from ASR and to tentatively identify the active metabolites in ASR using non-targeted metabolomics. Based on the results of the independent variables test, the interaction effects of three key extracting variables, including methanol concentration, ultrasonic time, and material-to-liquid ratio, were further investigated using the Box–Behnken design (BBD) to obtain prior active components with high AChE inhibitory activity. UPLC-QTOF-MS combined with a multivariate method was used to analyze the metabolites in ASR and investigate the causes of activity differences. Based on the current study, the optimal conditions for UAE were as follows: methanol concentration of 85.06%, ultrasonic time of 39.1 min, and material-to-liquid ratio of 1.06:10 (g/mL). Under these optimal conditions, the obtained extracts show strong inhibitions against AChE with half maximal inhibitory concentration (IC50) values ranging from 0.375 to 0.459 µg/mL according to an Ellman’s method evaluation. Furthermore, 55 metabolites were identified from the ASR extracted using methanol in different concentrations, and 9 biomarkers were subsequently identified as potential compounds responsible for the observed AChE inhibition. The active extracts have potential to be used for the development of functional foods with positive effects on Alzheimer’s disease owing to their high AChE inhibition activity. Altogether, this study provides insights into promoting the comprehensive utilization of A. truncatum resources

Fucosylated Human Milk Oligosaccharides during the First 12 Postnatal Weeks Are Associated with Better Executive Functions in Toddlers

Human milk oligosaccharides (HMOs) are one of the most abundant solid components in a mother's milk. Animal studies have confirmed a link between early life exposure to HMOs and better cognitive outcomes in the offspring. Human studies on HMOs and associations with later child cognition are scarce. In this preregistered longitudinal study, we investigated whether human milk 2'-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, grouped fucosylated HMOs, and grouped sialylated HMOs, assessed during the first twelve postnatal weeks, are associated with better child executive functions at age three years. At infant age two, six, and twelve weeks, a sample of human milk was collected by mothers who were exclusively (n = 45) or partially breastfeeding (n = 18). HMO composition was analysed by use of porous graphitized carbon-ultra high-performance liquid chromatography-mass spectrometry. Executive functions were assessed at age three years with two executive function questionnaires independently filled in by mothers and their partners, and four behavioural tasks. Multiple regression analyses were performed in R. Results indicated that concentrations of 2'-fucosyllactose and grouped fucosylated HMOs were associated with better executive functions, while concentrations of grouped sialylated HMOs were associated with worse executive functions at age three years. Future studies on HMOs that sample frequently during the first months of life and experimental HMO administration studies in exclusively formula-fed infants can further reveal associations with child cognitive development and uncover potential causality and sensitive periods

Preparation and Characterization of Phenolic Acid-Chitosan Derivatives as an Edible Coating for Enhanced Preservation of Saimaiti Apricots

In this study, caffeic acid (CA) and chlorogenic acid (CGA) were incorporated onto chitosan (CS) using free radical grafting initiated by a hydrogen peroxide/ascorbic acid (H2O2/Vc) redox system. The structural properties of the CA (CA-g-CS) and CGA (CGA-g-CS) derivatives were characterized by UV&ndash;Vis absorption, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), nuclear magnetic resonance (NMR), and thermal stability analysis. Then, the antioxidant and antibacterial properties were evaluated, and the effect of CGA-g-CS on the postharvest quality of Saimaiti apricot was studied. It proved that phenolic acids were successfully grafted onto the CS. The grafting ratios of CA-g-CS and CGA-g-CS were 126.21 mg CAE/g and 148.94 mg CGAE/g. The antioxidation and antibacterial activities of CGA-g-CS were better than those of CA-g-CS. The MICs of CGA-g-CS against E. coli, S. aureus, and B. subtilis were 2, 1, and 2 mg/mL. The inhibitory zones of 20 mg/mL CGA-g-CS against the three bacteria were 19.16 &plusmn; 0.35, 16.33 &plusmn; 0.91, and 16.24 &plusmn; 0.05 mm. The inhibitory effects of 0.5% CGA-g-CS on the firmness, weight loss, SSC, TA, relative conductivity, and respiration rate of the apricot were superior. Our results suggest that CGA-g-CS can be potentially used as an edible coating material to preserve apricots