RESUMEN
Raffinose family oligosaccharides (RFOs) have diverse structures and exhibit various biological activities. When using RFOs as prebiotics, their structures need to be identified. If we first knew whether an RFO was classical or non-classical, structural identification would become much easier. Here, we cloned and expressed an α-galactosidase (BF0224) from Bacteroides fragilis which showed strict specificity for hydrolyzing α-Gal-(1 â 6)-Gal linkages in RFOs. BF0224 efficiently distinguished classical from non-classical RFOs by identifying the resulting hydrolyzed oligo- and mono-saccharides with HPAEC-PAD-MS. Using this strategy, we identified a non-classical RFO from Pseudostellaria heterophylla (Miquel) Pax with DP6 (termed PHO-6), as well as a classical RFO from Lycopus lucidus Turcz. with DP7 (termed LTO-7). To characterize these RFO structures, we employed four other commercial or reported α-galactosidases in combination with NMR and methylation analysis. Using this approach, we elucidated the accurate chemical structure of PHO-6 and LTO-7. Our study provides an efficient analytical approach to structurally analyze RFOs. This enzyme-based strategy also can be applied to structural analysis of other glycans.
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Bacteroides fragilis , Oligosacáridos , Rafinosa , alfa-Galactosidasa , Bacteroides fragilis/enzimología , alfa-Galactosidasa/química , alfa-Galactosidasa/metabolismo , alfa-Galactosidasa/genética , Rafinosa/química , Rafinosa/metabolismo , Oligosacáridos/química , HidrólisisRESUMEN
Even though Stellaria dichotoma L. var. lanceolate (S. dichotoma) is a well-known medicinal plant in the family Caryophyllaceae, its oligosaccharides remain unexplored in terms of their potential as bioactive agents. Here, we isolated a mixture of oligosaccharides from S. dichotoma (Yield: 12 % w/w), that are primarily non-classical raffinose family oligosaccharides (RFOs). Nine major oligosaccharides were purified and identified from the mixture, including sucrose, raffinose, 1-planteose, lychnose, stellariose, along with four new non-classical RFOs. Two of the four new oligosaccharides are linear hexose pentamers with α-galactosyl extensions on their lychnose moieties, and the other two are branched hexose hexamers with α-galactosyl extensions on their stellariose groups. Their interactions with galectin-3 (Gal-3) revealed significant binding, with the terminal galactose providing enhanced affinity for the lectin. Notably, Gal-3 residues Arg144, His158, Asn160, Arg162, Asn174, Trp181, Glu184 and Arg186 coordinate with the lychnose. In vivo studies using the dextran sulfate sodium (DSS) mouse model for colitis demonstrated the ability of these carbohydrates in mitigating ulcerative colitis (UC). Overall, our study has provided structural information and potential applications of S. dichotoma oligosaccharides, also offers new approaches for the development of medicinal oligosaccharides.
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Colitis , Galectina 3 , Oligosacáridos , Animales , Oligosacáridos/química , Oligosacáridos/farmacología , Ratones , Galectina 3/metabolismo , Galectina 3/química , Colitis/tratamiento farmacológico , Colitis/inducido químicamente , Colitis/metabolismo , Caryophyllaceae/química , Sulfato de Dextran , Ratones Endogámicos C57BL , Masculino , HumanosRESUMEN
Lacto-N-neotetraose (LNnT), as a neutral core structure within human milk oligosaccharides (HMOs), has garnered widespread attention due to its exceptional physiological functions. In the process of LNnT synthesis using cellular factory approaches, substrate promiscuity of glycosyltransferases leads to the production of longer oligosaccharide derivatives. Here, rational modification of ß1,3-N-acetylglucosaminyltransferase from Neisseria meningitidis (LgtA) effectively decreased the concentration of long-chain LNnT derivatives. Specifically, the optimal ß1,4-galactosyltransferase (ß1,4-GalT) was selected from seven known candidates, enabling the efficient synthesis of LNnT in Escherichia coli BL21(DE3). Furthermore, the influence of lactose concentration on the distribution patterns of LNnT and its longer derivatives was investigated. The modification of LgtA was conducted with computational assistance, involving alanine scanning based on molecular docking to identify the substrate binding pocket and implementing large steric hindrance on crucial amino acids to obstruct LNnT entry. The implementation of saturation mutagenesis at positions 223 and 228 of LgtA yielded advantageous mutant variants that did not affect LNnT synthesis while significantly reducing the production of longer oligosaccharide derivatives. The most effective mutant, N223I, reduced the molar ratio of long derivatives by nearly 70 %, showcasing promising prospects for LNnT production with diminished byproducts.
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N-Acetilglucosaminiltransferasas , Neisseria meningitidis , Oligosacáridos , Neisseria meningitidis/enzimología , N-Acetilglucosaminiltransferasas/metabolismo , N-Acetilglucosaminiltransferasas/genética , Oligosacáridos/química , Oligosacáridos/síntesis química , Simulación del Acoplamiento Molecular , Escherichia coli/genética , Especificidad por Sustrato , Lactosa/análogos & derivados , Lactosa/metabolismo , Lactosa/química , HumanosRESUMEN
The use of antibiotics as conventional feed additives in poultry operations have proven useful, however resulted serious health concerns to consumer due to their bio-accumulation, besides rising problem of antimicrobial resistance in microbes, thus, an alternative to antibiotic growth promoter have called for. One of the aim of the experiment was to assess the lone and combined effects of feeding of chitosan oligosaccharide (COS) and blend of organic acids and short chain fatty acids in essential oils on growth performance, haematological parameters, relative lymphoid organ weight and innate immunity in early aged layer chick (male birds). A total of ninety, day-old chicks were randomly allotted into five groups: CO, Control group fed only poultry feed ; AGP, antibiotic growth promoter fed Avilomycin at the dose of 200 mg/kg of poultry feed; CH, chitosan oligosaccharide fed at the rate of 100 mg/kg feed; OE, blend of organic acids and short chain fatty acids in essential oils contained 1000 to 2000 mg/kg feed in a graded dose per week and CH + OE, chitosan oligosaccharide plus blend of organic acids and short chain fatty acids in essential oils at consistent rate and manner as followed for each of given feed additives when fed individually. Data on growth performance, samples for haematological parameters and innate immunity were measured and assayed on 7th, 21st and 42nd day post feeding (dpf) respectively. The results showed that compared with the control group; there is a marginal gain in body weight at 7th and 21st dpf in CH group and the corresponding CH + OE group. Feed conversion ratio in CH group was remarkably good at 7th and 21st dpf. No significant difference was observed in relative organ weights of thymus, spleen and Bursa of Fabricius in treatment groups as compared to control birds; however a significant rise in splenic weight index in OE fed birds at 42nd dpf noted. Haematological changes were inconsequential in treatment groups with an exception to enhancement of heterophil to lymphocyte ratio (H:L ratio) in CH group at 42nd dpf. Serum lysozyme activity proportionately elevated in CH + OE group on 21st and 42nd dpf when measured against control group; on the other hand no detectable augmentation of gut lysozyme activity observed. Both serum bactericidal and gut bactericidal activity boosted in combinatorial group at 42nd dpf. These results indicated that early age feeding of chitosan individually or combination with organic acids and short chain fatty acids in layer chick is beneficial, as it has the potential to enhance body weight gain to some extent and improves systemic and localized innate immunity to offer protection against infectious assaults thus may avoid early chick mortality in farms.
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Alimentación Animal , Pollos , Quitosano , Inmunidad Innata , Animales , Quitosano/administración & dosificación , Quitosano/farmacología , Pollos/crecimiento & desarrollo , Pollos/inmunología , Masculino , Inmunidad Innata/efectos de los fármacos , Alimentación Animal/análisis , Dieta/veterinaria , Suplementos Dietéticos/análisis , Distribución Aleatoria , Aceites Volátiles/administración & dosificación , Aceites Volátiles/farmacología , Tamaño de los Órganos/efectos de los fármacos , Oligosacáridos/administración & dosificación , Oligosacáridos/farmacologíaRESUMEN
BACKGROUND: Individuals with non-celiac gluten/wheat sensitivity (NCGWS) experience improvement in gastrointestinal symptoms following a gluten-free diet. Although previous results have indicated that fructo-oligosaccharides (FOS), a type of short-chain fructans, were more likely to induce symptoms than gluten in self-reported NCGWS patients, the underlying mechanisms are unresolved. METHODS: Our main objective was therefore to investigate whether FOS-fructans and gluten affect the composition and diversity of the faecal microbiota (16S rRNA gene sequencing), faecal metabolites of microbial fermentation (short-chain fatty acids [SCFA]; gas chromatography with flame ionization detector), and a faecal biomarker of gut inflammation (neutrophil gelatinase-associated lipocalin, also known as lipocalin 2, NGAL/LCN2; ELISA). In the randomised double-blind placebo-controlled crossover study, 59 participants with self-reported NCGWS underwent three different 7-day diet challenges with gluten (5.7 g/day), FOS-fructans (2.1 g/day), and placebo separately (three periods, six challenge sequences). RESULTS: The relative abundances of certain bacterial taxa were affected differently by the diet challenges. After the FOS-fructan challenge, Fusicatenibacter increased, while Eubacterium (E.) coprostanoligenes group, Anaerotruncus, and unknown Ruminococcaceae genera decreased. The gluten challenge was primarily characterized by increased abundance of Eubacterium xylanophilum group. However, no differences were found for bacterial diversity (α-diversity), overall bacterial community structure (ß-diversity), faecal metabolites (SCFA), or NGAL/LCN2. Furthermore, gastrointestinal symptoms in response to FOS-fructans were generally not linked to substantial shifts in the gut bacterial community. However, the reduction in E. coprostanoligenes group following the FOS-fructan challenge was associated with increased gastrointestinal pain. Finally, correlation analysis revealed that changes in gastrointestinal symptoms following the FOS-fructan and gluten challenges were linked to varying bacterial abundances at baseline. CONCLUSIONS: In conclusion, while FOS-fructans induced more gastrointestinal symptoms than gluten in the NCGWS patients, we did not find that substantial shifts in the composition nor function of the faecal microbiota could explain these differences in the current study. However, our results indicate that individual variations in baseline bacterial composition/function may influence the gastrointestinal symptom response to both FOS-fructans and gluten. Additionally, the change in E. coprostanoligenes group, which was associated with increased symptoms, implies that attention should be given to these bacteria in future trials investigating the impact of dietary treatments on gastrointestinal symptoms. TRIAL REGISTRATION: Clinicaltrials.gov as NCT02464150.
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Estudios Cruzados , Heces , Fructanos , Microbioma Gastrointestinal , Glútenes , Humanos , Masculino , Femenino , Microbioma Gastrointestinal/efectos de los fármacos , Microbioma Gastrointestinal/fisiología , Glútenes/efectos adversos , Glútenes/administración & dosificación , Adulto , Heces/microbiología , Heces/química , Persona de Mediana Edad , Método Doble Ciego , Hipersensibilidad al Trigo/dietoterapia , Oligosacáridos/administración & dosificación , Adulto JovenRESUMEN
Breastfeeding provides many health benefits, but its impact on respiratory health remains unclear. This study addresses the complex and dynamic nature of the mother-milk-infant triad by investigating maternal genomic factors regulating human milk oligosaccharides (HMOs), and their associations with respiratory health among human milk-fed infants. Nineteen HMOs are quantified from 980 mothers of the CHILD Cohort Study. Genome-wide association studies identify HMO-associated loci on chromosome 19p13.3 and 19q13.33 (lowest P = 2.4e-118), spanning several fucosyltransferase (FUT) genes. We identify novel associations on chromosome 3q27.3 for 6'-sialyllactose (P = 2.2e-9) in the sialyltransferase (ST6GAL1) gene. These, plus additional associations on chromosomes 7q21.32, 7q31.32 and 13q33.3, are replicated in the independent INSPIRE Cohort. Moreover, gene-environment interaction analyses suggest that fucosylated HMOs may modulate overall risk of recurrent wheeze among preschoolers with variable genetic risk scores (P < 0.01). Thus, we report novel genetic factors associated with HMOs, some of which may protect the respiratory health of children.
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Estudio de Asociación del Genoma Completo , Leche Humana , Oligosacáridos , Sialiltransferasas , Humanos , Leche Humana/química , Leche Humana/metabolismo , Femenino , Oligosacáridos/metabolismo , Sialiltransferasas/genética , Sialiltransferasas/metabolismo , Lactante , Masculino , Preescolar , Fucosiltransferasas/genética , Lactancia Materna , Ruidos Respiratorios/genética , Interacción Gen-Ambiente , Polimorfismo de Nucleótido Simple , Adulto , Estudios de Cohortes , Madres , Niño , Cromosomas Humanos Par 3/genética , Lactosa/análogos & derivadosRESUMEN
We present the application of N-difluoroacetylglucosamine (GlcNDFA) in a chemical evolution strategy to synthesize oligosaccharides. In comparison to conventional N-trifluoroacetylglucosamine, GlcNDFA exhibits superior substrate compatibility with glycosyltransferases as well as stability in aqueous environments. Using our 16-step assembly line, GlcNDFA can be used to produce homogeneous dekaparin, a heparin-like medication, with a yield of 62.2%. This underscores the significant potential of GlcNDFA as a chemical evolution precursor in the precise synthesis of structurally defined polysaccharides.
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Glicosiltransferasas , Glicosilación , Estructura Molecular , Glicosiltransferasas/metabolismo , Glicosiltransferasas/química , Hexosaminas/química , Hexosaminas/síntesis química , Oligosacáridos/química , Oligosacáridos/síntesis químicaRESUMEN
BACKGROUND: Human milk oligosaccharides (HMOs), which are unique bioactive components in human milk, are increasingly recognized for their multifaceted roles in infant health. A deeper understanding of the nexus between HMOs and the gut-brain axis can revolutionize neonatal nutrition and neurodevelopmental strategies. METHODS: We performed a narrative review using PubMed, Embase, and Google Scholar to source relevant articles. The focus was on studies detailing the influence of HMOs on the gut and brain systems, especially in neonates. Articles were subsequently synthesized based on their exploration into the effects and mechanisms of HMOs on these interconnected systems. RESULTS: HMOs significantly influence the neonatal gut-brain axis. Specific concentrations of HMO, measured 1 and 6 months after birth, would seem to agree with this hypothesis. HMOs are shown to influence gut microbiota composition and enhance neurotransmitter production, which are crucial for brain development. For instance, 2'-fucosyllactose has been demonstrated to support cognitive development by fostering beneficial gut bacteria that produce essential short-chain fatty acids. CONCLUSIONS: HMOs serve as crucial modulators of the neonatal gut-brain axis, underscoring their importance in infant nutrition and neurodevelopment. Their dual role in shaping the infant gut while influencing brain function presents them as potential game-changers in neonatal health strategies.
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Eje Cerebro-Intestino , Microbioma Gastrointestinal , Fenómenos Fisiológicos Nutricionales del Lactante , Leche Humana , Oligosacáridos , Humanos , Leche Humana/química , Microbioma Gastrointestinal/fisiología , Eje Cerebro-Intestino/fisiología , Recién Nacido , Fenómenos Fisiológicos Nutricionales del Lactante/fisiología , Encéfalo/metabolismo , Encéfalo/crecimiento & desarrollo , Desarrollo Infantil , Lactante , Femenino , TrisacáridosRESUMEN
A diet with low content of fermentable oligo-, di-, and monosaccharides and polyols (FODMAP) is established treatment for irritable bowel syndrome (IBS), with well-documented efficiency. A starch- and sucrose-reduced diet (SSRD) has shown similar promising effects. The primary aim of this randomized, non-inferiority study was to test SSRD against low FODMAP and compare the responder rates (RR = ∆Total IBS-SSS ≥ -50) to a 4-week dietary intervention of either diet. Secondary aims were to estimate responders of ≥100 score and 50% reduction; effects on extraintestinal symptoms; saturation; sugar craving; anthropometric parameters; and blood pressure. 155 IBS patients were randomized to SSRD (n = 77) or low FODMAP (n = 78) for 4 weeks, with a follow-up 5 months later without food restrictions. The questionnaires Rome IV, IBS-severity scoring system (IBS-SSS), and visual analog scale for IBS (VAS-IBS) were completed at baseline and after 2 and 4 weeks and 6 months. Weight, height, waist circumference, and blood pressures were measured. Comparisons were made within the groups and between changes in the two groups. There were no differences between groups at baseline. The responder rate of SSRD was non-inferior compared with low FODMAPs at week 2 (79.2% vs. 73.1%; p = 0.661;95% confidence interval (CI) = -20-7.2) and week 4 (79.2% vs. 78.2%; p = 1.000;95%CI = -14-12). Responder rate was still high when defined stricter. All gastrointestinal and extraintestinal symptoms were equally improved (p < 0.001 in most variables). SSRD rendered greater reductions in weight (p = 0.006), body mass index (BMI) (p = 0.005), and sugar craving (p = 0.05), whereas waist circumference and blood pressure were equally decreased. Weight and BMI were regained at follow-up. In the SSRD group, responders at 6 months still had lowered weight (-0.7 (-2.5-0.1) vs. 0.2 (-0.7-2.2) kg; p = 0.005) and BMI (-0.25 (-0.85-0.03) vs. 0.07 (-0.35-0.77) kg/m2; p = 0.009) compared with baseline in contrast to non-responders. Those who had tested both diets preferred SSRD (p = 0.032). In conclusion, a 4-week SSRD intervention was non-inferior to low FODMAP regarding responder rates of gastrointestinal IBS symptoms. Furthermore, strong reductions of extraintestinal symptoms were found in both groups, whereas reductions in weight, BMI, and sugar craving were most pronounced following SSRD.
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Síndrome del Colon Irritable , Almidón , Humanos , Síndrome del Colon Irritable/dietoterapia , Femenino , Masculino , Adulto , Persona de Mediana Edad , Almidón/administración & dosificación , Resultado del Tratamiento , Fermentación , Polímeros , Monosacáridos , Dieta Baja en Carbohidratos/métodos , Sacarosa en la Dieta/administración & dosificación , Oligosacáridos , Disacáridos/administración & dosificación , Presión SanguíneaRESUMEN
Changes in dietary patterns and living habits have led to an increasing number of individuals with elevated cholesterol levels. Excessive consumption of high-cholesterol foods can disrupt the body's lipid metabolism. Numerous studies have firmly established the cholesterol-lowering effects of probiotics and prebiotics, with evidence showing that the synergistic use of synbiotics is functionally more potent than using probiotics or prebiotics alone. Currently, the screening strategy involves screening prebiotics for synbiotic development with probiotics as the core. However, in comparison to probiotics, there are fewer types of prebiotics available, leading to limited resources. Consequently, the combinations of synbiotics obtained are restricted, and probiotics and prebiotics are only relatively suitable. Therefore, in this study, a novel synbiotic screening strategy with prebiotics as the core was developed. The synbiotic combination of Lactobacillus rhamnosus S_82 and xylo-oligosaccharides was screened from the intestinal tract of young people through five generations of xylo-oligosaccharides. Subsequently, the cholesterol-lowering ability of the medium was simulated, and the two carbon sources of glucose and xylo-oligosaccharides were screened out. The results showed that synbiotics may participate in cholesterol-lowering regulation by down-regulating the expression of NPC1L1 gene, down-regulating ACAT2 and increasing the expression of ABCG8 gene in vitro through cell adsorption and cell absorption in vitro, and regulating the intestinal microbiota. Synbiotics hold promise as potential candidates for the prevention of hypercholesterolemia in humans and animals, and this study providing a theoretical foundation for the development of new synbiotic products.
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Lacticaseibacillus rhamnosus , Oligosacáridos , Prebióticos , Simbióticos , Lacticaseibacillus rhamnosus/metabolismo , Oligosacáridos/farmacología , Humanos , Hipolipemiantes/farmacología , Colesterol/metabolismo , Colesterol/sangre , Microbioma Gastrointestinal/efectos de los fármacos , Probióticos , GlucuronatosRESUMEN
The purpose of this study was to investigate the potential prebiotic properties of cassava cultivars from Northeast [Doce mel and Ourinho (OUR)] and South [Baiana, and IPR-Upira (UPI)] of Brazil in in vitro fermentation systems. The cultivars were evaluated for their chemical composition, and, then, two cultivars were selected (OUR and UPI) and subjected to in vitro gastrointestinal digestion to assess the effects on probiotics Lacticaseibacillus casei, Lactobacillus acidophilus, and Bifidobacterium animalis growth, metabolic activity, and prebiotic activity scores. Finally, the impact of cassava cultivars on the fecal microbiota of celiac individuals was evaluated using the 16S rRNA gene. Cassava cultivars have variable amounts of fiber, resistant starch, fructooligosaccharides (FOS), organic acids, phenolic compounds, and sugars, with OUR and UPI cultivars standing out. OUR and UPI cultivars contributed to the increase in the proliferation rates of L. casei (0.04-0.19), L. acidophilus (0.34-0.27), and B. animalis (0.10-0.03), resulting in more significant effects than FOS, an established prebiotic compound. Also, the positive scores of prebiotic activities with probiotic strains indicate OUR and UPI's ability to stimulate beneficial bacteria while limiting enteric competitors selectively. In addition, OUR and UPI promoted increased relative abundance of Bifidobacteriaceae, Enterococcaceae, and Lactobacillaceae in the fecal microbiota of celiac individuals while decreased Lachnospirales, Bacteroidales, and Oscillospirales. The results show that cassava cultivars caused beneficial changes in the composition and metabolic activity of the human intestinal microbiota of celiacs. OUR and UPI cultivars from the Northeast and South of Brazil could be considered potential prebiotic ingredients for use in the formulation of functional foods and dietary supplements.
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Enfermedad Celíaca , Heces , Fermentación , Microbioma Gastrointestinal , Manihot , Prebióticos , Manihot/química , Humanos , Brasil , Heces/microbiología , Enfermedad Celíaca/dietoterapia , Enfermedad Celíaca/microbiología , Colon/microbiología , Colon/metabolismo , Lactobacillus acidophilus , Masculino , Probióticos , Adulto , ARN Ribosómico 16S/genética , Femenino , Oligosacáridos , Lacticaseibacillus casei , Bifidobacterium animalisRESUMEN
Breast milk is widely acknowledged as the ideal nutritional resource for infants and can well meet the nutritional requirements for baby's growth and development. Infant formula is a substitute for breast milk, designed to closely mimic its composition and function for breast milk. Most of the previous studies used tumor colorectal cancer cell lines to study the nutritional potency of formula and its components, so realistic data closer to the baby could not be obtained. Small intestinal organoids, derived from differentiated human embryonic stem cells, can be used to simulate nutrient absorption and metabolism in vitro. In this experiment, we used small intestinal organoids to compare the nutrient absorption and metabolism of three infant formulae for 0-6 months with breast milk samples. Transcriptome and metabolome sequencing methods were used to analyze the differentially expressed genes (DEGs) and differentially expressed metabolites (DEMs). The pathways related to DEGs, DEMs were enriched using GO, KEGG, GSEA and other methods to investigate their biological characteristics. We have found that both formula and breast milk promote the development of the infant's immune system, nutrient absorption and intestinal development. In PMH1 we found that the addition of oligofructose to milk powder promoted lipid metabolism and absorption. In PMH2 we found that whey protein powder favours the development of the immune system in infants. In PMH3 we found that oligogalactans may act on the brain-gut axis by regulating the intestinal flora, thereby promoting axon formation and neural development. By linking these biological properties of the milk powder with its composition, we confirmed the effects of added ingredients on the growth and development of infants. Also, we demonstrated the validity of small intestine organoids as a model for absorption and digestion in vitro. Through the above analyses, the advantages and disadvantages of the roles of formula and breast milk in the growth and metabolism of infants were also compared.
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Fórmulas Infantiles , Intestino Delgado , Metaboloma , Leche Humana , Organoides , Transcriptoma , Humanos , Leche Humana/metabolismo , Leche Humana/química , Organoides/metabolismo , Intestino Delgado/metabolismo , Intestino Delgado/citología , Lactante , Oligosacáridos/metabolismo , Recién Nacido , Absorción Intestinal , Femenino , Proteína de Suero de Leche/metabolismoRESUMEN
There are great challenges in the field of natural product isolation and purification and in the pharmacological study of oligosaccharide monomers. And these isolation and purification processes are still universal problems in the study of natural products (NPs), traditional Chinese medicine (TCM), omics, etc. The same polymer-modified materials designed for the special separation of oligosaccharides, named Sil-epoxy-PEI and Sil-chloropropyl-PEI, were synthesized via two different methods and characterized by scanning electron microscopy combined with energy spectrum analysis, Fourier transform infrared spectroscopy, thermogravimetric analysis, zeta potential as well as surface area analysis, etc. Several nucleotide/nucleoside molecules with different polarities and selectivities were successfully isolated in our laboratory using stainless-steel columns filled with the synthesized material. In addition, the separation of saccharide probes and oligosaccharides mixtures in water extracts of Morinda officinalis were compared in HILIC mode. The results showed that the resolution of separations for the representative analytes of the Sil-epoxy-PEI column was higher than for the Sil-chloropropyl-PEI column, and the developed stationary phase exhibited improved performance compared to hydrothermal carbon, amide columns and other HILIC materials previously reported.
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Oligosacáridos , Polietileneimina , Dióxido de Silicio , Oligosacáridos/química , Oligosacáridos/síntesis química , Oligosacáridos/aislamiento & purificación , Polietileneimina/química , Dióxido de Silicio/química , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Immunoglobulin A (IgA) is a major gut antibody that coats commensal gut bacteria and contributes to shaping a stable gut bacterial composition. Although previous studies have shown that cyclic oligosaccharides, including cyclic nigerosyl-1,6-nigerose (CNN) and cyclodextrins (CDs, including αCD, ßCD, and γCD), alter the gut bacterial composition, it remains unclear whether cyclic oligosaccharides modify the IgA coating of gut bacteria, which relates to cyclic oligosaccharide-induced alteration of the gut bacterial composition. To address this issue, mice were maintained for 12 weeks on diets containing CNN, αCD, ßCD, or γCD; the animals' feces were evaluated for their bacterial composition and the IgA coating index (ICI), a measure of the degree of IgA coating of bacteria. We observed that the intake of each cyclic oligosaccharide altered the gut bacterial composition, with changes in the ICI found at both the phylum and genus levels. The ICI for Bacillota, Lachnospiraceae NK4A136 group, UC Lachnospiraceae, and Tuzzerella were significantly and positively correlated with the relative abundance (RA) in total bacteria for these bacteria; in contrast, significant correlations were not seen for other phyla and genera. Our observations suggest that cyclic oligosaccharide-induced modulation of the IgA coating of gut bacteria may partly relate to changes in the community structure of the gut bacteria.
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Heces , Microbioma Gastrointestinal , Inmunoglobulina A , Oligosacáridos , Animales , Microbioma Gastrointestinal/efectos de los fármacos , Oligosacáridos/farmacología , Ratones , Heces/microbiología , Bacterias/efectos de los fármacos , Masculino , Ciclodextrinas/farmacología , Ratones Endogámicos C57BLRESUMEN
Early-life nutrition significantly impacts vaccination efficacy in infants, whose immune response to vaccines is weaker compared to adults. This study investigated vaccination efficacy in female C57Bl/6JOlaHsd mice (6 weeks old) fed diets with 0.7% galacto-oligosaccharides (GOS)/long-chain fructo-oligosaccharides (lcFOS) (9:1), 0.3% human milk oligosaccharides (HMOS), or a combination (GFH) for 14 days prior to and during vaccination. Delayed-type hypersensitivity (DTH) was measured by assessing ear swelling following an intradermal challenge. Influvac-specific IgG1 and IgG2a levels were assessed using ELISAs, while splenic T and B lymphocytes were analyzed for frequency and activation via flow cytometry. Additionally, cytokine production was evaluated using murine splenocytes co-cultured with influenza-loaded dendritic cells. Mice on the GFH diet showed a significantly enhanced DTH response (p < 0.05), increased serological IgG1 levels, and a significant rise in memory B lymphocytes (CD27+ B220+ CD19+). GFH-fed mice also exhibited more activated splenic Th1 cells (CD69+ CXCR3+ CD4+) and higher IFN-γ production after ex vivo restimulation (p < 0.05). These findings suggest that GOS/lcFOS and HMOS, particularly in combination, enhance vaccine responses by improving memory B cells, IgG production, and Th1 cell activation, supporting the potential use of these prebiotics in infant formula for better early-life immune development.
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Vacunas contra la Influenza , Ratones Endogámicos C57BL , Leche Humana , Oligosacáridos , Animales , Oligosacáridos/farmacología , Leche Humana/inmunología , Leche Humana/química , Femenino , Vacunas contra la Influenza/inmunología , Humanos , Ratones , Vacunación , Inmunoglobulina G/sangre , Galactosa , Linfocitos B/inmunología , Bazo/inmunología , Citocinas/metabolismo , Modelos Animales de Enfermedad , Anticuerpos Antivirales/sangreRESUMEN
Human milk, the gold standard in infant nutrition, is a unique fluid that provides essential nutrients such as lactose, lipids, proteins, and free oligosaccharides. While its primary role is nutritional, it also protects against pathogens. This protection mainly comes from immunoglobulins, with human milk oligosaccharides (HMOs) providing additional support by inhibiting pathogen binding to host cell ligands. The prebiotic and immune-modulatory activity of HMOs strongly depends on their structure. Over 200 individual structures have been identified so far, with the composition varying significantly among women. The structure and composition of HMOs are influenced by factors such as the Lewis blood group, secretor status, and the duration of nursing. HMO profiles are heavily influenced by maternal phenotypes, which are defined based on the expression of two specific fucosyltransferases. However, recent data have shown that HMO content can be modified by various factors, both changeable and unchangeable, including diet, maternal age, gestational age, mode of delivery, breastfeeding frequency, and race. The first part of this overview presents the historical background of these sugars and the efforts by scientists to extract them using the latest chromatography methods. The second part is divided into subchapters that examine modifiable and non-modifiable factors, reviewing the most recent articles on HMO composition variations due to specific reasons and summarizing potential future challenges in conducting these types of studies.
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Leche Humana , Oligosacáridos , Leche Humana/química , Humanos , Oligosacáridos/análisis , Femenino , Lactancia Materna , Antígenos del Grupo Sanguíneo de Lewis , Prebióticos , DietaRESUMEN
Less than half of all patients diagnosed with pancreatic ductal adenocarcinoma (PDAC) respond to chemotherapy, and the prognosis of PDAC is poor, which may be mediated by the gut microbiota. We investigated the clinical improvement effects of 1-kestose, a fructooligosaccharide, on PDAC chemotherapy in this single-center, randomized, controlled pilot trial conducted at Fujita Health University Hospital, which enrolled patients with PDAC. The trial included 1-kestose administration and non-administration groups. The 1-kestose group received 9 g of 1-kestose daily for 12 weeks, and their blood markers, imaging studies, physical findings, and gut microbiota were evaluated. In the 1-kestose administration group, the cancer marker CA19-9 significantly decreased, and there was a reduction in the neutrophil-to-lymphocyte ratio (NLR). There was also suppression of the reduction of albumin levels and of an increase in C-reactive protein. Additionally, Escherichia coli, which typically increases in PDAC, significantly decreased in the 1-kestose group. Thus, 1-kestose altered the gut microbiota and improved the prognostic factors for PDAC. Large-scale, long-term trials of 1-kestose interventions for PDAC are thus warranted to improve the prognosis of PDAC.
Asunto(s)
Carcinoma Ductal Pancreático , Microbioma Gastrointestinal , Neoplasias Pancreáticas , Humanos , Proyectos Piloto , Carcinoma Ductal Pancreático/tratamiento farmacológico , Masculino , Femenino , Neoplasias Pancreáticas/tratamiento farmacológico , Microbioma Gastrointestinal/efectos de los fármacos , Anciano , Persona de Mediana Edad , Suplementos Dietéticos , Biomarcadores de Tumor/sangre , Pronóstico , Antígeno CA-19-9/sangre , Proteína C-Reactiva/análisis , Proteína C-Reactiva/metabolismo , Escherichia coli/efectos de los fármacos , Resultado del Tratamiento , Neutrófilos , Oligosacáridos/administración & dosificación , Oligosacáridos/farmacologíaRESUMEN
Cello-oligosaccharides (COS) become a new type of functional oligosaccharides. COS transglycosylation reactions were studied to enhance COS yield production. Seeking the ability of the free form of Fusarium solani ß-glucosidase (FBgl1) to synthesize COS under low substrate concentrations, we found out that this biocatalyst initiates this reaction with only 1 g/L of cellobiose, giving rise to the formation of cellotriose. Cellotriose and cellopentaose were detected in biphasic conditions with an immobilized FBgl1 and when increased to 50 g/L of cellobiose as a starter concentration. After the biocatalyst recycling process, the trans-glycosylation yield of COS was maintained after 5 cycles, and the COS concentration was 6.70 ± 0.35 g/L. The crude COS contained 20.15 ± 0.25 g/L glucose, 23.15 ± 0.22 g/L non-reacting substrate cellobiose, 5.25 ± 0.53 g/L, cellotriose and 1.49 ± 0.32 g/L cellopentaose. A bioprocess was developed for cellotriose enrichment, using whole Bacillus velezensis cells as a microbial purification tool. This bacteria consumed glucose, unreacted cellobiose, and cellopentaose while preserving cellotriose in the fermented medium. This study provides an excellent enzyme candidate for industrial COS production and is also the first study on the single-step COS enrichment process.
Asunto(s)
Bacillus , Celobiosa , Fusarium , Oligosacáridos , beta-Glucosidasa , Fusarium/enzimología , Fusarium/metabolismo , Fusarium/genética , beta-Glucosidasa/metabolismo , Oligosacáridos/metabolismo , Celobiosa/metabolismo , Bacillus/enzimología , Bacillus/metabolismo , Bacillus/genética , Prebióticos , Glicosilación , Glucosa/metabolismoRESUMEN
BACKGROUND: Immunomodulatory proteins in human milk (HM) can shape infant immune development. However, strategies to modulate their levels are currently unknown. This study investigated whether maternal prebiotic supplementation alters the levels of immunomodulatory proteins in HM. METHODS: The study was nested within the SYMBA double-blind randomized controlled trial (ACTRN12615001075572), which investigated the effects of maternal prebiotic (short-chain galacto-oligosaccharides/long-chain fructo-oligosaccharides) supplementation from <21 weeks gestation during pregnancy until 6 months postnatal during lactation on child allergic disease risk. Mother-child dyads receiving prebiotics (n = 46) or placebo (n = 54) were included in this study. We measured the levels of 24 immunomodulatory proteins in HM collected at 2, 4, and 6 months. RESULTS: Cluster analysis showed that the overall immunomodulatory protein composition of milk samples from both groups was similar. At 2 months, HM of prebiotic-supplemented women had decreased levels of TGF-ß1 and TSLP (95% CI: -17.4 [-29.68, -2.28] and -57.32 [-94.22, -4.7] respectively) and increased levels of sCD14 (95% CI: 1.81 [0.17, 3.71]), when compared to the placebo group. At 4 months, IgG1 was lower in the prebiotic group (95% CI: -1.55 [-3.55, -0.12]) compared to placebo group. CONCLUSION: This exploratory study shows that prebiotic consumption by lactating mothers selectively alters specific immunomodulatory proteins in HM. This finding is crucial for understanding how prebiotic dietary recommendations for pregnant and lactating women can modify the immune properties of HM and potentially influence infant health outcomes through immune support from breastfeeding.
Asunto(s)
Suplementos Dietéticos , Leche Humana , Prebióticos , Humanos , Leche Humana/inmunología , Leche Humana/química , Prebióticos/administración & dosificación , Femenino , Método Doble Ciego , Embarazo , Lactante , Adulto , Masculino , Lactancia/inmunología , Oligosacáridos/administración & dosificación , Recién Nacido , Lactancia Materna , Citocinas/metabolismoRESUMEN
OBJECTIVES: We hypothesized that a low fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAP) diet may be associated with the disease severity of ulcerative colitis (UC). Therefore, this cross-sectional study was undertaken to assess the association between FODMAP score and UC severity. METHODS: In this study, 158 patients with UC were enrolled. The disease severity was designated using the Mayo score. The diet relation information was obtained using the 160-item food frequency questionnaire. To calculate the FODMAP score, the consumption of all food items was converted to a gram per day and multiplied by the FODMAP factor. The FODMAP factor was obtained from the application developed by Monash University (Melbourne, Victoria, Australia). The association between disease severity (dependent factor) and FODMAP score tertiles (independent factors) was assessed by logistic regression adjusted for different covariates. RESULTS: In the present study, the age range of participants was 18 to 64 y old, and 46.2% of patients had moderate or severe disease activity. There were significant differences in sex, body mass index, and supplement use across different tertiles of FODMAP score. There was no significant association between the FODMAP score tertiles and disease severity in the crude model and adjusted models. CONCLUSIONS: The results of the present study showed that there was no significant association between the FODMAP score and UC severity. However, considering the limitations of the study, more studies with prospective and interventional designs using more accurate methods of dietary assessments are needed to confirm these preliminary results.