Levels Of Oligosaccharides Research Articles

Mannan oligosaccharides improve the fur quality of raccoon dogs by regulating the gut microbiota.

Adding antibiotics to animal basal diets can improve growth and production performance. However, the use of antibiotics poses a potential threat to public health safety. The study was conducted to investigate the effects of different levels of mannan oligosaccharides (MOS) on the fur quality, nutrient apparent digestibility, serum immunity, antioxidant status, intestinal morphology, and gut microbiota of fur-growing raccoon dogs. Divide 24 male raccoon dogs (120 ± 5 d) of similar weight (5.01 ± 0.52 kg) into 4 groups randomly. Add 0, 0.05, 0.1, and 0.2% MOS to the basal diets of groups C, L, M, and H, respectively. Compared to the C group, the addition of 0.05% and 0.1% MOS in the diet increased the apparent digestibility of crude protein (CP), Underfur length (UL), Guard hair length (GL), immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) levels in the serum (p < 0.05); Under the dosage of 0.05 % MOS, the activities of Superoxide Dismutase (SOD) and catalase (CAT) increased (p < 0.05). Compared to the C group, adding 0.05% MOS significantly increased the VH/CD of the duodenum and ileum, while also increasing the VH and CD of the jejunum (p < 0.05). Through Spearman correlation analysis of the gut microbiota, it was found that MOS can improve fur quality by reducing the abundance of Dorea while improving the immune response of raccoon dogs by reducing the abundance of Blautia and Gemmiger. In conclusion, MOS can improve the fur quality, serum immunity, antioxidant capacity, and gut microbiota of raccoon dogs. Therefore, MOS has the potential to replace antibiotics.

A platform for qualitative and quantitative characterization of α-Gal and NeuGc at the oligosaccharide level

Glycosylation modification serves as a pivotal quality attribute in glycoprotein-based therapeutics, emphasizing its role in drug safety and efficacy. Prior studies have underscored the potential immunogenic risks posed by the presence of galactose-α-1,3-galactose (α-Gal) and N-glycolylneuraminic acid (NeuGc) in glycoprotein formulations. This accentuates the imperative for developing robust qualitative and quantitative analytical methods to monitor these immunogenic epitopes, thereby ensuring drug safety. In the present investigation, α-Gal and NeuGc were accurately quantified using UPLC-FLR-MS/MS at the oligosaccharide level. Remarkably, α-Gal could be identified when the ion intensity ratio or the mass-to-charge ratio (m/z) of 528.19 to 366.14 exceeded 1. Concurrently, NeuGc and N-acetylneuraminic acid (NeuAc) could be unambiguously identified through their respective fragment ions at m/z 673.23 and m/z 657.23. Furthermore, relative quantification of α-Gal and NeuGc was achieved using fluorescence signals. This study introduces a sensitive and reliable analytical approach for the qualitative and quantitative assessment of α-Gal and NeuGc in glycoprotein pharmaceuticals. The methodology offers significant potential for application in process control and optimization of glycoprotein production, aimed at minimizing immunogenicity and enhancing product quality.

Alterations of milk oligosaccharides in mothers with gestational diabetes mellitus impede colonization of beneficial bacteria and development of RORγt+ Treg cell-mediated immune tolerance in neonates

ABSTRACT Gestational diabetes mellitus (GDM) is an increasing public health concern that significantly increases the risk of early childhood allergic diseases. Altered maternal milk glycobiome may strongly affect gut microbiota and enteric-specific Treg cell-mediated development of immune tolerance in GDM infants. In this study, we found that, compared with healthy Chinese mothers, mothers with GDM had significantly lower levels of total and specific human milk oligosaccharides (HMOs) in their colostrum that subsequently increased with extension of lactation. This alteration in HMO profiles significantly delayed colonization of Lactobacillus and Bifidobacterium spp. in their breast-fed infants, resulting in a distinct gut microbial structure and metabolome. Further experiments in GDM mouse models indicated that decreased contents of milk oligosaccharides, mainly 3ʹ-sialyllactose (3ʹ-SL), in GDM maternal mice reduced colonization of bacteria, such as L. reuteri and L. johnsonii, in the neonatal gut, which impeded development of RORγt+ regulatory T (Treg) cell-mediated immune tolerance. Treatment of GDM neonates with 3ʹ-SL, Lactobacillus reuteri (L. reuteri) and L. johnsonii promoted the proliferation of enteric Treg cells and expression of transcription factor RORγt, which may have contributed to compromising ovalbumin (OVA)-induced allergic responses. In vitro experiments showed that 3ʹ-SL, metabolites of L. johnsonii, and lysates of L. reuteri stimulated differentiation of mouse RORγt+ Treg cells through multiple regulatory effects on Toll-like receptor, MAPK, p53, and NOD-like receptor signaling pathways. This study provides new ideas for the development of gut microbiota and immune tolerance in GDM newborns.

Effects of Functional Oligosaccharides Extracted from Straw on the Growth, Feeding, Physiology, Histology, Muscle Texture, and Gut Microbiota of Micropterus salmoides

We conducted an 8-week feeding trial to investigate the effect of mixed functional oligosaccharides (chitosan oligosaccharide, fructooligosaccharide, and xylooligosaccharide) extracted from wheat straw on the growth, feeding, physiology, histology, muscle texture, and gut microbiota of Micropterus salmoides. Six diets were formulated by incrementally adding mixed functional oligosaccharides (0.1, 0.5, 1.0, 1.5, and 2.0 mg/kg) to the control diet. 30 individuals with initial body weight of 25.1 ± 3.5 g were randomly allocated to 6 triplicate tanks and fed to apparent satiation twice daily. The fish fed 1.0 mg/kg additives displayed maximum growth, and the feed conversion ratio decreased with the increase in additives from 0 to 1.0 mg/kg ( P &lt; 0.05 ) but did not decline further with the addition of more than 1.0 mg/kg ( P &gt; 0.05 ). The villus height and width were significantly higher in the supplementation groups than in the control group, while the gut and liver structures presented abnormalities with excessive supplementation above 1.5 mg/kg. There were significant differences in muscle texture indices for M. salmoides over the fed additive gradient, and the hardness, gumminess, and chewiness were highest in the 1.0 mg/kg group. High oligosaccharide levels, such as 2.0 mg/kg, decreased the LZM level, while there were no significant differences in the SOD and MDA levels. Gut microbiome analysis revealed no significant differences in richness and diversity for groups fed the functional oligosaccharide gradient; however, the PCoA results showed that the microbial community composition changed markedly in response to different addition levels, and the 0.5 and 1.0 mg/kg supplementation groups were far apart from the lower and higher supplementation groups. The relative abundance of Proteobacteria was lower in the 0.5 and 1.0 mg/kg addition groups, while that of the phyla Fusobacteria and Firmicutes were higher in these two groups. Functional classification showed that microbes related to carbohydrate metabolism were more abundant in the 0.5 and 1.5 mg/kg groups than in the other groups.

The Application of HPLC-FLD and NMR in the Monitoring of Therapy Efficacy in Alpha-Mannosidosis.

Alpha-mannosidosis is a rare lysosomal storage disorder, caused by decreased activity of α-D-mannosidase. This enzyme is involved in the hydrolysis of mannosidic linkages in N-linked oligosaccharides. Due to the mannosidase defect, undigested mannose-rich oligosaccharides (Man2GlcNAc - Man9GlcNAc) accumulating in cells are excreted in large quantities in urine. In this work, we determined the levels of urinary mannose-rich oligosaccharides in a patient subjected to novel enzyme replacement therapy. Urinary oligosaccharides were extracted using solid phase extraction (SPE), labeled by fluorescent tag 2-aminobenzamide, and quantified by high-performance liquid chromatography (HPLC) with fluorescence detector (FLD). The identity of peaks was determined by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry. In addition, the levels of urinary mannose-rich oligosaccharides were also quantified by 1H nuclear magnetic resonance (NMR) spectroscopy. The data were analyzed using one-tailed paired t-test and Pearson's correlation tests. Compared to levels before the administration of therapy, an approximately two-folds decrease in total mannose-rich oligosaccharides after one month of treatment was observed by NMR and HPLC. After four months, an approximately ten-folds significant decrease in total urinary mannose-rich oligosaccharides was detected, suggesting therapy effectiveness. A significant decrease in the levels of oligosaccharides with 7-9 mannose units was detected by HPLC. The application of both HPLC-FLD and NMR in quantification of oligosaccharide biomarkers is a suitable approach for monitoring of therapy efficacy in alpha-mannosidosis patients.

Dynamic Changes in the Human Milk Metabolome Over 25 Weeks of Lactation.

Human milk (HM) provides essential nutrition for ensuring optimal infant growth and development postpartum. Metabolomics offers insight into the dynamic composition of HM. Studies have reported the impact of lactation stage, maternal genotype, and gestational age on HM metabolome. However, the majority of the studies have considered changes within the first month of lactation or sampled with large intervals. This leaves a gap in the knowledge of progressing variation in HM composition beyond the first month of lactation. The objective of this study was to investigate whether the HM metabolome from mothers with term deliveries varies beyond 1 month of lactation, during the period in which HM is considered fully mature. Human milk samples (n = 101) from 59 mothers were collected at weeks 1–2, 3–5, 7–9, and 20–25 postpartum and analyzed using 1H nuclear magnetic resonance spectroscopy. Several metabolites varied over lactation and exhibited dynamic changes between multiple time points. Higher levels of HM oligosaccharides, cis-aconitate, O-phosphocholine, O-acetylcarnitine, gluconate, and citric acid were observed in early lactation, whereas later in lactation, levels of lactose, 3-fucosyllactose, glutamine, glutamate, and short- and medium-chain fatty acids were increased. Notably, we demonstrate that the HM metabolome is dynamic during the period of maturity.

Periodate oxidation of plant polysaccharides provides polysaccharide-specific oligosaccharides

Although polysaccharides are frequently used in foods, detailed characterization and/or identification of their structures using a single method remains a challenge. We investigated the suitability of periodate oxidation as an approach to depolymerize polysaccharides, allowing characterization and/or identification of the original polysaccharides based on ESI-MS analyses of the released oligosaccharides. Various periodate oxidation conditions were tested on (arabino)xylan, galactomannan, xyloglucan and homogalacturonan. Each polysaccharide required a different oxidation condition to release a substantial level of oligosaccharides. These oligosaccharides had highly complex structures due to the presence of e.g., dialdehyde sugars, hemialdals, and remnants of (oxidized) sugars, as verified by ESI-MS/MS. Despite these oligosaccharides were highly complex and lost some polysaccharide structural features, each periodate-oxidized sample comprised polysaccharide structure-dependent MS oxidized oligosaccharide profiles. Our findings are a good starting point to find a more generic chemical polysaccharide depolymerization approach based on periodate oxidation to identify polysaccharides by oligosaccharides fingerprinting using MS.

Corn arabinoxylan has a repeating structure of subunits of high branch complexity with slow gut microbiota fermentation

Corn arabinoxylan (CAX), a cell wall-derived dietary fiber, was extracted with alkali, partially purified, and treated with hydrolytic enzymes in order to investigate the relationship of fine structure and fermentability by the human gut microbiota. Glycosyl composition and linkage analysis of CAX and two hydrolysates, coupled with molecular size analysis, indicated an organized structural feature of the native polymer, which consists of a repeating structural subunit containing complex branching patterns along the xylan backbone and flanked by regions of less complexity. The two lengths of the highly branched subunit were isolated and were shown to have enhanced slow fermentation property compared to the native structure (3.3 vs. 5.9 mL gas, 4 h), that was related to increasing complexity of the branched structures. Lower molecular size structures with higher branch complexity fermented slower, contrary to a conventional view that small fiber structures approaching the oligosaccharide level are necessarily more rapidly fermented.

A Survey to Identify the Current Management of Cow's Milk Disorders and the Role of Goat Milk-Based Formulas in the Middle East and North Africa Region.

Background: Cow’s milk allergy (CMA) and cow’s milk intolerance (CMI) are the major cow’s milk disorders observed in infants and young children. This study investigates, for the first time, physician knowledge regarding CMA and CMI prevalence, diagnosis, and management in the Middle East and North Africa (MENA) region. In addition, we explore the role of goat milk-based formula as an alternative in infants suffering from CMI. Method: This cross-sectional survey was conducted from December 2020 to February 2021. A convenience sample of 2500 MENA-based physicians received the questionnaire, developed by a working group of pediatric experts. Results: 1868 physicians completed the questionnaire, including pediatric specialists (80.8%), training physicians (0.2%), dermatologists (0.1%), family/general physicians (12.9%), neonatologists (3.6%), neurosurgeons (0.2%), allergy nurse specialists (0.3%), pharmacists (2.1%), and public health workers (0.1%). Differentiation between CMA and CMI was recognized by the majority of respondents (80.7%), for which the majority of respondents (35.4%) identified that the elimination and challenge test was the best test to differentiate CMA from CMI, whereas 30.7% and 5.4% preferred the immunoglobulin E (IgE) test and skin prick test, respectively. In addition, 28.5% of respondents reported that there is no confirmatory test to differentiate CMA from CMI. The majority of respondents (47.3%) reported that amino acid-based formula (AAF)/ extensively hydrolyzed formula (EHF) is the cornerstone for the management of CMA. However, most respondents (33.7%) reported that lactose avoidance was best for the management of CMI. Overall, 65% of the respondents were aware of nutritionally adapted goat’s milk formula as an alternative to cow’s milk products and 37% would recommend its routine use in infants (≤2 years of age). Conclusion: The results of this survey demonstrate that the majority of physicians are aware of the underlying pathophysiology and management of CMA and CMI. However, a significant proportion of physicians do not follow the clinical guidelines concerning CMA/CMI diagnosis and management. Notably, this survey identified that goat’s milk formulas may offer a suitable alternative to AAF/EHF in infants with CMI as they contain β-casein protein which is easily digestible. In addition, goat’s milk formulas contain higher levels of oligosaccharides and medium-chained fatty acids compared with standard cow’s milk formulas, yet further clinical trials are warranted to support the inclusion of goat’s milk formulas in clinical guidelines.

Desiccation as a Post-maturation Treatment Helps Complete Maturation of Norway Spruce Somatic Embryos: Carbohydrates, Phytohormones and Proteomic Status.

Exposure of Norway spruce (Picea abies) somatic embryos and those of many other conifers to post-maturation desiccation treatment significantly improves their germination. An integration analysis was conducted to understand the underlying processes induced during the desiccation phase at the molecular level. Carbohydrate, protein and phytohormone assays associated with histological and proteomic studies were performed for the evaluation of markers and actors in this phase. Multivariate comparison of mature somatic embryos with mature desiccated somatic embryos and/or zygotic embryos provided new insights into the processes involved during the desiccation step of somatic embryogenesis. Desiccated embryos were characterized by reduced levels of starch and soluble carbohydrates but elevated levels of raffinose family oligosaccharides. Desiccation treatment decreased the content of abscisic acid and its derivatives but increased total auxins and cytokinins. The content of phytohormones in dry zygotic embryos was lower than in somatic embryos, but their profile was mostly analogous, apart from differences in cytokinin profiles. The biological processes “Acquisition of desiccation tolerance”, “Response to stimulus”, “Response to stress” and “Stored energy” were activated in both the desiccated somatic embryos and zygotic embryos when compared to the proteome of mature somatic embryos before desiccation. Based on the specific biochemical changes of important constituents (abscisic acid, raffinose, stachyose, LEA proteins and cruciferins) induced by the desiccation treatment and observed similarities between somatic and zygotic P. abies embryos, we concluded that the somatic embryos approximated to a state of desiccation tolerance. This physiological change could be responsible for the reorientation of Norway spruce somatic embryos toward a stage suitable for germination.

Biochemical Evaluation of Moong Bean (Vigna radiata L.) Characterstics for Nutraceutical Applications

Background: Moong bean (Vigna radiate L.) plays an important role in antioxidant, antimicrobial, antidiabetic, anti inflammatory, antihypertensive and antitumor effects. Mostly beans and legumes cause gas and abdominal bloating in some people but the moong beans are the easy to digest. Soaking the beans in water before cooking and also sprouting them reduces this issue. The high levels of amino acids, oligosaccharides and polyphenols provide high antioxidant levels not only fight tumors and cancer but also protect against DNA damage and cell mutation. The seed of moong bean acts as a nutraceutical because it provides beneficial effects on human health and also helps in the prevention of cardiovascular disease, diabetes, digestive tract diseases, overweight, obesity and cancer etc. The flavonoids vitexin and isovitexin have high free radical scavenging activities which help lower oxidative stress, the cancer causative factor. It is not only lower blood glucose levels, but also reduce total cholesterol levels, triglycerides, plasma C peptides and glucagon levels. Samples of moong bean were collected from Legume Section of the University. Methods: Through NIRS-2500 evaluated samples of moong bean. It is an instrument through which Near Infrared (NIR) analysis a spectroscopic technique that makes use of the naturally occurring electromagnetic spectrum. This works in the region of the spectrum defined by wavelengths between 700 nm and 2500 nm. All the cultivars were found to cluster in major four groups on the basis of principal component analysis. The result showed the diversity between nutritional and antinutritional factors in the cultivars that could be further used by plant breeders to develop superior genotypes. The moong bean has lots of advantages which cure the several diseases such as cardiovascular disease, type 2 diabetes, digestive diseases and some cancers. Result: Histidine amino acid was varied from 6.40% to 8.81%, arginine amino acid from 2.21% to 3.87%, valine amino acid from 2.92% to 4.44%, phenylalanine from 9.73% to 13.86%, isoleucine from 9.28% to 10.02%, methionine from 0.09% to 1.26%, leucine from 6.02% to 7.65%, tryptophan from 0.30% to 0.68%, lysine from 2.03% to 2.22% and threonine from 4.32% to 5.56%. The grain color have the direct related to carotenoids and polyphenols. The hardness of the grain is due to fiber amount of the grain. The moisture is related to the properties such as sphericity, porosity, bulk density. Mostly anti-nutrients factors such as phytic acid, tannins, hemagglutinins and polyphenols associated with the grain. The effects of storage and processing on nutrients and food processing properties are needed to enable optimization of processing steps, for better mung bean food quality and process efficiency.

Oligosaccharide Presentation Modulates the Molecular Recognition of Glycolipids by Galectins on Membrane Surfaces.

Galectins are a family of glycan binding proteins that stand out for the wide range of biological phenomena in which they are involved. Most galectin functions are associated with their glycan binding capacities, which are generally well characterized at the oligosaccharide level, but not at the glycoprotein or glycolipid level. Glycolipids form the part of cell membranes where they can act as galectin cellular receptors. In this scenario, glycan presentation as well as the membrane chemical and structural features are expected to have a strong impact in these molecular association processes. Herein, liposomes were used as membrane mimicking scaffolds for the presentation of glycosphingolipids (GSLs) and to investigate their interaction with Galectin-3 and the N-domain of Galectin-8 (Gal8N). The binding towards GM3 and GM1 and their non-silaylated GSLs was compared to the binding to the free glycans, devoid of lipid. The analysis was carried out using a combination of NMR methods, membrane perturbation studies, and molecular modeling. Our results showed a different tendency of the two galectins in their binding capacities towards the glycans, depending on whether they were free oligosaccharides or as part of GSL inserted into a lipid bilayer, highlighting the significance of GSL glycan presentation on membranes in lectin binding.

Association Between Breastmilk Microbiota and Food Allergy in Infants.

Regulating the composition of human breastmilk has the potential to prevent allergic diseases early in life. The composition of breastmilk is complex, comprising varying levels of oligosaccharides, immunoactive molecules, vitamins, metabolites, and microbes. Although several studies have examined the relationship between different components of breastmilk and infant food allergies, few have investigated the relationship between microorganisms in breastmilk and infant food allergy. In the present study, we selected 135 healthy pregnant women and their full-term newborns from a cohort of 202 mother–infant pairs. Among them, 69 infants were exclusively breastfed until 6 mo after birth. At follow-up, 11 of the 69 infants developed a food allergy in infancy while 22 showed no signs of allergy. Thirty-three breastmilk samples were collected within 1 mo after delivery, and 123 infant fecal samples were collected at five time points following their birth. These samples were analyzed using microbial 16S rRNA gene sequencing. The abundance and evenness of the milk microbiota and the number of differential bacteria were higher in the breastmilk samples from the non-allergy group than in those from the food allergy group. The non-allergy group showed relatively high abundance of Bifidobacterium, Akkermansia, Clostridium IV, Clostridium XIVa, Veillonella, and butyrate-producing bacteria such as Fusobacterium, Lachnospiraceae incertae sedis, Roseburia, and Ruminococcus. In contrast, the abundance of Proteobacteria, Acinetobacter, and Pseudomonas in breastmilk was higher in the food allergy group. A comparison of the changes in dominant differential breastmilk microbiota in the intestinal flora of the two groups of infants over time revealed that the changes in Bifidobacterium abundance were consistent with those in the breastmilk flora. Functional pathway prediction of breastmilk microflora showed that the enhancement of the metabolic pathways of tyrosine, tryptophan, and fatty acids was significantly different between the groups. We suggest that changes in the breastmilk microbiota can influence the development of food allergies. Breastmilk contains several microbes that have protective effects against food allergies, both by influencing the colonization of intestinal microbiota and by producing butyrate. This study may provide new ideas for improving infant health through early intervention with probiotics.

Short-term germination of faba bean (Vicia faba L.) and the effect on selected chemical constituents

Selected chemical compounds of five faba bean (FB) cultivars; Snowdrop (low tannin), Taboar (high tannin), Snowbird (low tannin), Fabelle (low vicine & convicine), and Malik (high tannin) that were grown in North America (3 production years) were determined. The effect of short-term germination (24 h, 48 h, and 72 h) on the levels of phytic acid, vicine/convicine (v/cv), oligosaccharides, and total phenolics were evaluated. On average, FB contained 2% phytic acid and there was no difference between the 5 cultivars. Fabelle presented the lowest level of v/cv of 1 and 0.4 mg/g, respectively while the others had 7.27–10.82 and 3.53–3.93 mg/g, respectively. No significant difference was found among cultivars for oligosaccharides content. Content of total phenolics differed significantly (p ≤ 0.05) between FB cultivars. Snowdrop and Snowbird had the highest total phenolic content of 48.4 and 48.3 mg/g, respectively, while Fabelle had the lowest level of 10.8 mg/g. All cultivars showed significantly (p ≤ 0.05) reduced contents of raffinose (100%), stachyose (60%), verbascose (80%) at 72 h. Germination had no significant effect on phytic acid, vicine (except Snowbird), and convicine levels. At 48-h, Snowdrop and Snowbird seeds only showed 17-18% reduction (p ≤ 0.05) in total phenolic content. Germination between 48 to 72 h is an option to produce FB seeds with very low levels of oligosaccharides.

Six Oligosaccharides' Variation in Breast Milk: A Study in South China from 0 to 400 Days Postpartum.

This study investigated the variation in oligosaccharide levels in the breast milk of south Chinese mothers in a prolonged breastfeeding period of up to 400 days postpartum. A total of 488 breast milk samples were collected from 335 healthy mothers at five different time points: 0–5 days, 10–15 days, 40–45 days, 200–240 days, and 300–400 days postpartum. A high-performance anion-exchange chromatography-pulsed amperometric detector (HPAEC-PAD) was used to quantify 2′-fucosyllactose (2′-FL), 3-fucosyllactose (3-FL), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 3′-sialyllactose (3′-SL) and 6′-sialyllactose (6′-SL). In this study, we found six oligosaccharides that were present in breast milk from 0 to 400 days postpartum. The median value ranges of individual oligosaccharide components in this study were 1013–2891 mg/L 2′-FL, 193–1421 mg/L 3-FL, 314–1478 mg/L LNT, 44–255 mg/L LNnT, 111–241 mg/L 3′-SL, and 23–602 mg/L6′-SL. HMO levels decreased over the lactation periods, except for 3-FL, which increased throughout lactation. The predominant fucosylated and sialylated HMOs were 2′-FL and 6′-SL at 40–45 days postpartum and changed to 3-FL and 3′-SL at 200–240 days postpartum. Results from this study showed that lactating women continue to provide their offspring with a high level of 2′-FL one year after delivery, suggesting that 2′-FL may play an important role for infants in early life. Our findings also provide further evidence in support of breastfeeding after one-year postpartum.

ROS-Scavengers, Osmoprotectants and Violaxanthin De-Epoxidation in Salt-Stressed Arabidopsis thaliana with Different Tocopherol Composition.

To determine the role of α- and γ-tocopherol (TC), this study compared the response to salt stress (200 mM NaCl) in wild type (WT) Arabidopsis thaliana (L.) Heynh. And its two mutants: (1) totally TC-deficient vte1; (2) vte4 accumulating γ-TC instead of α-TC; and (3) tmt transgenic line overaccumulating α-TC. Raman spectra revealed that salt-exposed α-TC accumulating plants were more flexible in regulating chlorophyll, carotenoid and polysaccharide levels than TC deficient mutants, while the plants overaccumulating γ-TC had the lowest levels of these biocompounds. Tocopherol composition and NaCl concentration affected xanthophyll cycle by changing the rate of violaxanthin de-epoxidation and zeaxanthin formation. NaCl treated plants with altered TC composition accumulated less oligosaccharides than WT plants. α-TC deficient plants increased their oligosaccharide levels and reduced maltose amount, while excessive accumulation of α-TC corresponded with enhanced amounts of maltose. Salt-stressed TC-deficient mutants and tmt transgenic line exhibited greater proline levels than WT plants, lower chlorogenic acid levels, and lower activity of catalase and peroxidases. α-TC accumulating plants produced more methylated proline- and glycine- betaines, and showed greater activity of superoxide dismutase than γ-TC deficient plants. Under salt stress, α-TC demonstrated a stronger regulatory effect on carbon- and nitrogen-related metabolites reorganization and modulation of antioxidant patterns than γ-TC. This suggested different links of α- and γ-TCs with various metabolic pathways via various functions and metabolic loops.

Lipoxygenase Enzymes, Oligosaccharides (Raffinose and Stachyose) and 11sA4 and A5 Globulins of Glycinin Present in Soybean Meal Are Not Drivers of Enteritis in Juvenile Atlantic Salmon (Salmo salar)

Soybean meal has been largely investigated and commercially used in fish nutrition. However, its inclusion levels have been carefully considered due to the presence of antinutritional factors, which depending on a series of factors might induce gut inflammation damaging the mucosal integrity and causing enteritis. Several strategies including genetic engineering have been applied attempting to reduce or eliminate some of the antinutritional factors. Accordingly, we assessed the intestinal health of juvenile Atlantic salmon fed high levels of speciality soybean genotypes with reduced-to-no content amounts of lipoxygenases, altered glycinin profile and reduced levels of oligosaccharides. No major signs of enteritis, only indication of enteritis progression, was noticed in the soybean meal-based diets illustrated by mild changes in distal intestine morphology. Whereas fish, fed fishmeal control feeds, displayed normal distal intestine integrity. Speciality soybean types did not improve intestinal health of juvenile Atlantic salmon suggesting these antinutrients are not drivers of the intestinal inflammatory process in this species. No additional benefits in terms of production performance or blood biochemistry were noticed in the speciality soybean types compared to the traditional soybean.

Levels of human milk oligosaccharides in breast milk of mothers delivering preterm infants of different gestational ages and their effects on early growth and development

Objective: To investigate the human milk oligosaccharides (HMOs) levels in breast milk of mothers delivering preterm infants and their effects on the early growth and development of infants. Methods: In this prospective cohort study, full-term and preterm newborns whose parents decided to breastfeed were recruited from Peking University Third Hospital between December 1, 2017 and November 30, 2018. The preterm infants were divided based on their gestational ages into extremely preterm (<28 weeks), very preterm (28-31+6 weeks) and moderate to late preterm (32-36+6 weeks) groups. Breast milk was collected from mothers at 7, 14, 28 and 120d postpartum. 368 breast milk samples were collected from 125 mothers in this study, including 54 mothers of full-term infants, 23 mothers of moderate to late preterm infants, 39 mothers of very preterm infants, and 9 mothers of extremely preterm infants. Ultra-performance liquid chromatography-mass spectrometer (UPLC-MS/MS) was used to determine the concentration of 2'-fucosyllactose (2'FL), 3-fucosyllactose (3FL), 3'-sialyllactose (3'SL), A-tetrasaccharide (P1), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-fucopentaose Ⅱ (LNFP-Ⅱ) and lacto-N-fucopentaose Ⅴ (LNFP-Ⅴ). Secretor status of mothers was defined as 2'-fucosyllactose (2'FL) concentration in colostrum and transitional milk greater than 200 μg/mL. Weight gain and the occurrence of allergic diseases of infants were collected at 120 d(4 months) postpartum. The chi-square test or the Fisher's exact test was used for the comparison of categorical data between groups; Kruskal-Wallis test and Wilcoxon rank sum test were used for comparison of continuous data between groups. Nemenyi test was used for multiple comparison. Results: 79.2% (99/125) of the mothers were secretor. There were no statistical differences between groups in the secretor status of mothers (χ²=1.31,P>0.05). The total concentration of HMOs peaked at 1-2 weeks postpartum. Compared to the preterm milk, the HMOs from the term milk was trending downwards at an earlier time. In the breast milk of secretor mothers on 28 d, total concentration of HMOs significant differed among the three groups of preterm milk and the term milk, with the median value of 4 587.09,4 615.25,5 277.44,5 476.03 μg/mL, respectively (Kruskal-Wallis χ²=8.1234,P=0.044). When analyzed by the median weight gain of the infants (low vs high weight gain) at 4 months postpartum, 2'FL was significantly lower in the high weight gain group at 7 d (1 818.04 μg/mL vs 2 181.67 μg/mL, W=1 386,P=0.018), while LNT & LNnT were significantly higher (1 182.36 μg/mL vs 1 053.62 μg/mL, W=816,P=0.044). The level of 3FL at 120 d was significantly affected by presence of allergic disease in infants, breast milk from mothers of infants with allergic disease had lower 3FL than those from mothers of infants without allergic disease (256.17 μg/mL vs 286.18 μg/mL, W=564,P=0.026). Conclusions: The overall profiles of HMOs in breast milk of mothers delivering preterm infants was basically the same as that of mothers delivering term infants; individual HMOs play a role in weight gain and the development of allergic diseases in preterm infants, but the mechanism is unclear and needs further study.

Presence and Levels of Galactosyllactoses and Other Oligosaccharides in Human Milk and Their Variation during Lactation and According to Maternal Phenotype.

Among the human milk oligosaccharides (HMOS), the galactosyllactoses (GLs) are only limitedly studied. This study aims to describe the presence and relative levels of HMOS, including GLs, in human milk (HM) according to maternal Secretor and Lewis (SeLe) phenotype and lactation stage. Relative levels of 19 HMOS were measured in 715 HM samples collected in the first 4 months postpartum from 371 donors participating in the PreventCD study. From a subset of 24 Dutch women (171 HM samples), samples were collected monthly up to 12 months postpartum and were additionally analyzed for relative and absolute levels of β6′-GL, β3′-GL and α3′-GL. Maternal SeLe phenotype or HM group was assigned based on the presence of specific fucosylated HMOS. Most HMOS, including β6′- and β3′-GL, were present in the vast majority (≥75%) of HM samples, whereas others (e.g., LNDFH II, 2′-F-LNH and α3′-GL) only occurred in a low number (<25%) of samples. Clear differences were observed between the presence and relative levels of the HMOS according to the maternal phenotype and lactation stage. Absolute concentrations of β6′-GL and β3′-GL were higher in HM group IV samples compared to samples of the other three HM groups. β3′-GL was also higher in HM group II samples compared to HM group I samples. β3′-GL and β6′-GL were stable over lactation stages. In conclusion, presence and levels of HMOS vary according to HM group and lactation stage. Not all HMOS behave similarly: some HMOS depend strongly on maternal phenotype and/or lactation stage, whereas others do not. β3′-GL and β6′-GL were present in low concentrations in over 75% of the analyzed HM samples and showed differences between HM groups, but not between the lactation stages.

Ultrasound treatment for improving the bioactive compounds and quality properties of a Brazilian nopal (Opuntia ficus-indica) beverage during shelf-life

The use of ultrasound treatment for improving the bioactive compounds and quality properties of a Brazilian nopal beverage was studied. Physicochemical characterization and possible changes in nutritional, microbiological, and sensory parameters throughout the shelf-life were evaluated, and compared with heat treatment. Five products were prepared and analyzed during 28 days of shelf-life: control without treatment (C0), beverage submitted to heat treatment for 10 min (HP10), and beverages submitted to ultrasound treatment for 10 (US10), 20 (US20), and 40 (US40) min, with these times being defined based on a previously conducted pilot study. The US40 beverage presented the highest levels of oligosaccharides (31.99%), organic (48.93%) and ascorbic (26.09%) acids, and minerals (83.59%) compared to those found for HP10. The US40 also showed better antioxidant activity and 81.80% more phenolic compounds when compared with the HP10. The US40 presented greater stability for bioactive compounds and showed fewer changes in the color parameters, lower microorganism count, and greater acceptability regarding quality during shelf-life, while the HP10 sample presented the worst results. Therefore, the US40 beverage has higher functional potential, and ultrasound represents a promising technique to preserve or improve the quality of this product during its processing and shelf-life.