Addition Of Oligosaccharides Research Articles

Short communication: Physicochemical and antioxidant properties of milk supplemented with red ginseng extract

This study investigated the effect of red ginseng extract (RGE) on the physicochemical properties, sensory test, and antioxidant activity of milk. The milk samples with RGE added at 0.5, 1, 1.5, and 2% were analyzed during storage at 4°C. The physicochemical properties included composition of milk, pH, titratable acidity, and color. The antioxidant activity of milk samples was determined using the 2,2-diphenyl-1-picrylhydrazyl method, β-carotene bleaching assay, and ferric thiocyanate assay. An increase in the amount of RGE in milk resulted in an increase of lactose and total solids content, titratable acidity, and a* and b* values, whereas fat and protein contents remained unchanged. Also, pH and L* value decreased. The antioxidant activity of milk samples supplemented with RGE was higher than that of the control sample. Sensory evaluation was performed using a quantitative descriptive analysis. Two types of samples were used: (1) sterilized milk fortified with RGE (0.5, 1, 1.5, and 2%) and (2) 2% RGE, 2% RGE with oligosaccharide, and 2% RGE with oligosaccharide and cyclodextrin. The addition of oligosaccharide and cyclodextrin could effect an increase of sweetness, a decrease of bitterness and flavor of RGE, and aftertaste. Therefore, milk supplemented with RGE could be useful as a functional food.

L-fucose from vitamin C with only acetonide protection.

Addition of human milk oligosaccharides (HMO) to baby foods may protect infants from disease. As many simple HMOs are fucosylated this is likely to increase the demand for L-fucose as a synthetic building block. Any chemical synthesis must be cheap to compete with a biotechnological process. Acetonide is the only protecting group we have used in this new synthesis of L-fucose from vitamin C in 27% overall yield (purification by recrystallization; no chromatography required in the entire sequence).

P.1.g.031 The neuroprotective and anti-amnestic effects of carbamylated erythropoietin derivates in ischemic brain injury

Purpose: To compare the neuroprotection of carbamylated erythropoietin (CEPO) and EPO fusion protein containing TR domain from glycoprotein MUC1 (CEPO-TR), and EPO fusion protein with modified Fc fragments of IgG1 (CEPO-Fc) against ischemic brain injury, including behavioral disturbances in a bilateral focal ischemic infarction of the rat prefrontal cortex. Methods: The carbamylated erythropoietin and erythropoietin derivates were produced by treatment of purified proteins with potassium cyanate in borate buffer. The resulting carbamylated erythropoietin and derivates exhibit no erythropoietic activity in UT-7/EPOR cell viability assay. Bilateral focal ischemic infarction of the prefrontal cortex (areas Frl and Fr2) was induced by the method of photochemical thrombosis in rats. After passive avoidance training and testing rats were treated, respectively, with following regimens: saline, 50mg/kg CEPO, 50mg/kg CEPO-TR, 50mg/kg CEPO-Fc. The substance was injected intraperitoneally in 1 h after operation. Neurological deficit scores and infarct volume were assessed at 4 and 7 days after operation. Functional state of CNS was determined by conditioned passive avoidance response (PA), i.e. by the latency of transition from light compartment to dark compartment (in sec). Morphometric measurements of the areas and volumes of the ischemic focus on serial slices were carried out on animal brain fixed by plunging into formalinethanol-acetic acid mixture (2:7:1). The data were statistically processed using Statistica 6.0 software. Summary of results: Additional oligosaccharides, linked to the erythropoietin, prolong its half-life and increase bioactivity in vivo. For this purpose we, for the first time, used TR domain from glycoprotein MUC1, bearing 5 additional sites for O-glycosylation. Our result indicated that ligation of TR-domain to the coding sequence of EPO did not affect secretion of the chimeric protein into the medium, receptor binding affinity in vitro bioactivity, compared with EPO wild type. However, both the in vitro potency and half-life in circulation of EPO bearing TR or Fc fragments were significantly enhanced. After photothrombosis and injection of saline, the latency of entry into the dark compartment decreased from 300 s to 76 s. Treatment of rats with CEPO, CEPOTR, and CEPO-Fc after photothrombosis restored latency of PA to 243, 258, 227 s, respectively, on day 4. Only injection of CEPOTR retains passive avoidance latency 207 s on the day 7. Ischemic damage volume in animals treated with CEPO-TR on 7th day after operation was 15.8±5.4 against 24.22±4.7 with control injection of saline (P< 0.05). This phenomenon on 7th day was observed only with CEPO-TR. Protection efficiency coefficient, calculated from these experiments represented 34%. Another erythropoietin derivate did not give statistically significant results. Conclusion: Treatment of rats with CEPO and CEPO fusion proteins after photothrombosis of cortex resulted in restoration of passive avoidance response and diminishing of volume of ischemic damage. Those, our study indicates that CEPO-Fc and CEPO-TR display neuroprotectionand anti-amnestic activity, while CEPO-TR demonstrates prolongation ability.

Comparison of Effects of Oligosaccharides on Physicochemical Properties of Corn Starch

Purpose: To investigate the effect of different oligosaccharides on the physicochemical properties of corn starch.Methods: The blue value and retrogradation of corn starch were evaluated following the addition of different oligosaccharides and compared with control. Pasting properties, melting enthalpy and melting temperature were determined by rapid viscosity analysis (RVA) and differential scanning calorimetry (DSC).Results: Maltose, lactose and fructo-oligosaccharides significantly affected iodine binding ability of corn starch. Retrogradation of corn starch was significantly inhibited by maltose, xylooligosaccharide and fructooligosaccharides. RVA analysis showed that the addition of oligosaccharides increased the peak viscosity of corn starch (from 3180 cP to 3687 cP) but decreased that of corn starch (from 896cP to 625 cP). DSC analysis showed that there was a large reduction in recrystallisation of corn starch by oligosaccharide. The samples containing xylooligosaccharide and fructooligosaccharides showed lower melting enthalpy (142.7 J/g and 143.5 J/g, respectively) than that of control (209.4 J/g), and their onset temperature (-10.9 °C and -8.89 °C, respectively) were also lower than that of control (-5.92 °C). In addition, the melting enthalpy and melting temperature of the samples containing oligosaccharides also showed lower values than those of control during storage.Conclusion: The finding that oligosaccharides modify the physicochemical properties of corn starch would be of practical importance in broadening their application prospects of the starch in the food and pharmaceutical industries.Keywords: Corn starch, Oligosaccharides, Pasting properties, Iodine binding, Retrogradation, Water activity, Enthalpy

Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics.

Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. The addition of oligosaccharides to recombinant proteins has been shown to greatly influence the overall physiochemical attributes of many proteins. It is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. The latter effect was demonstrated to increase antibody-dependent cell-mediated cytotoxicity for a recombinant antibody. These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability.

Can an ancestral condition for milk oligosaccharides be determined? Evidence from the Tasmanian echidna (Tachyglossus aculeatus setosus).

The monotreme pattern of egg-incubation followed by extended lactation represents the ancestral mammalian reproductive condition, suggesting that monotreme milk may include saccharides of an ancestral type. Saccharides were characterized from milk of the Tasmanian echidna Tachyglossus aculeatus setosus. Oligosaccharides in pooled milk from late lactation were purified by gel filtration and high-performance liquid chromatography using a porous graphitized carbon column and characterized by (1)H NMR spectroscopy; oligosaccharides in smaller samples from early and mid-lactation were separated by ultra-performance liquid chromatography and characterized by negative electrospray ionization mass spectrometry (ESI-MS) and tandem collision mass spectroscopy (MS/MS) product ion patterns. Eight saccharides were identified by (1)H NMR: lactose, 2'-fucosyllactose, difucosyllactose (DFL), B-tetrasaccharide, B-pentasaccharide, lacto-N-fucopentaose III (LNFP3), 4-O-acetyl-3'-sialyllactose [Neu4,5Ac(α2-3)Gal(β1-4)Glc] and 4-O-acetyl-3'-sialyl-3-fucosyllactose [Neu4,5Ac(α2-3)Gal(β1-4)[Fuc(α1-3)]Glc]. Six of these (all except DFL and LNFP3) were present in early and mid-lactation per ESI-MS, although some at trace levels. Four additional oligosaccharides examined by ESI-MS and MS/MS are proposed to be 3'-sialyllactose [Neu5Ac(α2-3)Gal(β1-4)Glc], di-O-acetyl-3'-sialyllactose [Neu4,5,UAc3(α2-3)Gal(β1-4)Glc where U = 7, 8 or 9], 4-O-acetyl-3'-sialyllactose sulfate [Neu4,5Ac(α2-3)Gal(β1-4)GlcS, where position of the sulfate (S) is unknown] and an unidentified 800 Da oligosaccharide containing a 4-O-acetyl-3'-sialyllactose core. 4-O-acetyl-3'-sialyllactose was the predominant saccharide at all lactation stages. 4-O-Acetylation is known to protect sialyllactose from bacterial sialidases and may be critical to prevent microbial degradation on the mammary areolae and/or in the hatchling digestive tract so that sialyllactose can be available for enterocyte uptake. The ability to defend against microbial invasion was probably of great functional importance in the early evolution of milk saccharides.

Biosynthesis and Export of Bacterial Lipopolysaccharides

Lipopolysaccharide molecules represent a unique family of glycolipids based on a highly conserved lipid moiety known as lipid A. These molecules are produced by most gram-negative bacteria, in which they play important roles in the integrity of the outer-membrane permeability barrier and participate extensively in host-pathogen interplay. Few bacteria contain lipopolysaccharide molecules composed only of lipid A. In most forms, lipid A is glycosylated by addition of the core oligosaccharide that, in some bacteria, provides an attachment site for a long-chain O-antigenic polysaccharide. The complexity of lipopolysaccharide structures is reflected in the processes used for their biosynthesis and export. Rapid growth and cell division depend on the bacterial cell's capacity to synthesize and export lipopolysaccharide efficiently and in large amounts. We review recent advances in those processes, emphasizing the reactions that are essential for viability.

Enhacement of the Viability of &amp;lt;i&amp;gt;Lactobacillus&amp;lt;/i&amp;gt; &amp;lt;i&amp;gt;plantarum&amp;lt;/i&amp;gt; during the Preservation and Storage Process Based on the Response Surface Methodology

Objective: The Response Surface Methodology (RSM) is a commonly used system to optimize cell viability of probiotic strains when they are subjected to different preservation and storage processes. Methods and Results: To determine the optimal levels of incorporation of several cry oprotectants (skim milk, sucrose and trehalose) in the freeze-drying process of Lactobacillus plantarum, a range of experiments based on a Rotational Central Composite design (CCD) were conducted. The results were adjusted to a quadratic model, resulting in the presence of interaction between the different variables. Solving a regression equation, we obtained the optimum concentrations of cryoprotective agents: 24.06% milk powder, 6.22% sucrose, 5.63% trehalose. To visualize the interactions between the three variables involved in the study, Design Expert? software was used. Conclusions: The analysis reveals that while trehalose has a direct effect on the viability of L. plantarum, skim milk and sucrose exert quadratic effects. There are also interactions between cryoprotectants, which emphasize the synergies produced between milk and sucrose and between sucrose and trehalose, which allows maintaining the viability of L. plantarum. Significance and Impact of the Study: The addition of new oligosaccharides as trehalose in premixtures for functional feed can maintain the viability of L. plantarum during longer periods of time, ensuring the proper administration of probiotics to their destinations.

Cyanidin-3-glucoside의 안정성을 향상시킨 오디즙 가공방법

오디의 기능성이 언론에 홍보됨에 따라 오디 소비 촉진 및 가공제품에 관한 관심이 증대되고 있으나 오디즙 가공시 기능성 물질인 C3G 색소의 안정성에 관한 기초자료가 없는 실정이다. 이러한 문제점을 해결하고 오디의 이용확대를 위해 항산화색소 C3G의 안정성을 향상시킨 오디즙 가공방법을 개발하였다. 그 결과를 요약하면 다음과 같다. 1. 오디와 서당의 비율을 1 : 1로 하여 용기에 담고 식품첨가제인 구연산을 0.3% 첨가하는 것이 C3G 함량과 항산화능을 향상시키는 가장 좋은 방법이다. 올리고당의 경우 구연산을 첨가하는 것이 오히려 항산화능을 떨어뜨리므로 사용하지 않는 것이 좋다. 자일리톨의 경우 가공방법 중 가장 낮은 C3G 함량을 나타냈으나 구연산을 첨가하면 색소함량과 항산화능을 향상시키므로 반드시 구연산을 첨가한다. 2. 보관용기(투명유리병, 갈색유리병, 알루미늄 호일, 녹색유리병, 반투명유리병)보다 구연산의 첨가가 항산화색소 C3G의 안정성에 영향을 미치는 것으로 판단된다. 3. 오디즙 가공시 전자렌지를 이용하여 3분간 blanching 하여 색소의 안정성에 관여하는 효소를 불활성화 시키는 방법은 항산화색소 C3G 함량을 다소 떨어뜨리므로 냉동오디를 그대로 사용하는 것이 바람직하나 항산화능에 있어서는 보존기간동안 타 처리구에 비해 안정화시키는 효과를 나타냈다. As mulberry fruit's functionalities are known to the media, the interest in promoting of consumption and processed products is increasing. But there is no C3G(Cyanidin-3-glucoside) data based on the stability of the pigment during processing. To solve this problems, and to expand the use of mulberry fruit, processing methods was developed for mulberry fruit juice improved cyanidin-3-glucoside(C3G) stability. The results obtained are summarized as follows. The food additive citric acid with 0.3 % improved C3G content and antioxidant ability in the treatment of mulberry fruit and sucrose the ratio of 50 % : 50 %(w/w). In the case of the addition of oligosaccharides, citric acid decreased antioxidant ability. Xylitol treatment showed up the lowest of C3 content, but by the addition of citric acid improved the pigment content and antioxidant activity of the mulberry fruit juice. Addition of citric acid was more affected C3G stability than containers (clear glass bottles, brown glass bottles, aluminum foil, green glass bottle, translucent glass bottles). In the processing of mulberry juice, 3 minutes blanching treatment using microwave dropped C3G content somewhat. Therefore, using this method is not recommended. However in the antioxidant ability, microwave blanching showed a stabilizing effect compared to the other treatments.

Recent advances in glycotechnology for glycoconjugate synthesis using microbial endoglycosidases

Biotechnology associated with synthesis of glycopeptides and glycoproteins has recently advanced as glycotechnology. Studies toward glycotechonology include the artificial modification of sugar chains in glycoconjugates to improve their function because the physiological importance of sugar chains in living organisms is well recognized. Methods involving addition of oligosaccharides to peptides and proteins have attracted attention as efficient techniques in glycotechnology, especially those involving the transglycosylation activities of microbial endoglycosidases. The exploration of oligosaccharide oxazolines as donor substrates for the transglycosylation of endoglycosidases has significantly enhanced the efficiency of these processes. Moreover, discovery of novel endoglycosidase mutants with glycosynthase-like activity has made it possible to effectively synthesize large quantities of glycopeptides, as well as homogeneous glycoprotein. The use of mutant enzymes and oligosaccharide oxazolines has led to development of practical applications for the synthesis of bioactive glycopeptides and therapeutic glycoproteins as bio-medicines.

Regulation of intracellular pH by glycosaminoglycans

The intracellular pH is regulated by a delicate balance of ion distribution across the plasma membrane and the physico-chemical properties of intra- and extracellular components. We analyzed the effects of glycosaminoglycans on the intracellular pH of fibroblasts by using the fluorescent pH indicator BCECF-AM. Addition of hyaluronan, hyaluronan oligosaccharides, chondroitin sulfate, or heparin to the culture medium of fibroblasts caused intracellular acidification from pH 7.2 to 6.7 in a concentration dependent manner. High molecular weight hyaluronan acidified more than hyaluronan oligosaccharides at the same concentrations. Hyaluronidase treatment or inhibition of hyaluronan export with xanthohumol led to intracellular alkalization. These observations indicated that extracellular glycosaminoglycans participate in intracellular pH regulation. The mechanism was explained by Donnan effects and molecular crowding.

Isolation of Sporothrix schenckii GDA1 and functional characterization of the encoded guanosine diphosphatase activity

Sporothrix schenckii is a fungal pathogen of humans and the etiological agent of sporotrichosis. In fungi, proper protein glycosylation is usually required for normal composition of cell wall and virulence. Upon addition of precursor oligosaccharides to nascent proteins in the endoplasmic reticulum, glycans are further modified by Golgi-glycosyl transferases. In order to add sugar residues to precursor glycans, nucleotide diphosphate sugars are imported from the cytosol to the Golgi lumen, the sugar is transferred to glycans, and the resulting nucleoside diphosphate is dephosphorylated by the nucleoside diphosphatase Gda1 before returning to cytosol. Here, we isolated the open reading frame SsGDA1 from a S. schenckii genomic DNA library. In order to confirm the function of SsGda1, we performed complementation assays in a Saccharomyces cerevisiae gda1∆ null mutant. Our results indicated that SsGDA1 restored the nucleotide diphosphatase activity to wild-type levels and therefore is a functional ortholog of S. cerevisiae GDA1.

Evidence for the Involvement of Intracellular Ca2+ Ions in the Elicitation Mechanism of Bacillus Licheniformis

Treatment of Bacillus licheniformis cultures with biotic oligosaccharide elicitors is known to increase the production of the antibiotic bacitracin A. The mechanism of the elicitation is currently under investigation and in this paper we provide evidence on the modulatory role of Ca²⁺ ions during this process. Addition of elicitors, mannan oligosaccharides, oligoguluronate and oligomannuronate to the liquid cultures resulted in 9.0, 5.2 and 5.0% increase in cytosolic Ca²⁺ levels in B. licheniformis, while the presence of verapamil (Ca²⁺ channel blocker) resulted in 74% decrease in bacitracin A levels, as compared to the control culture. We propose that Ca²⁺ ions may acts as a secondary messenger in the regulation of the bacitracin A synthesis in the elicited B. licheniformis cultures.

Uncovering Biphasic Catalytic Mode of C5-epimerase in Heparan Sulfate Biosynthesis

Heparan sulfate (HS), a highly sulfated polysaccharide, is biosynthesized through a pathway involving several enzymes. C(5)-epimerase (C(5)-epi) is a key enzyme in this pathway. C(5)-epi is known for being a two-way catalytic enzyme, displaying a "reversible" catalytic mode by converting a glucuronic acid to an iduronic acid residue, and vice versa. Here, we discovered that C(5)-epi can also serve as a one-way catalyst to convert a glucuronic acid to an iduronic acid residue, displaying an "irreversible" catalytic mode. Our data indicated that the reversible or irreversible catalytic mode strictly depends on the saccharide substrate structures. The biphasic mode of C(5)-epi offers a novel mechanism to regulate the biosynthesis of HS with the desired biological functions.

Structures of xyloglucans in primary cell walls of gymnosperms, monilophytes (ferns sensu lato) and lycophytes

Little is known about the structures of the xyloglucans in the primary cell walls of vascular plants (tracheophytes) other than angiosperms. Xyloglucan structures were examined in 13 species of gymnosperms, 13 species of monilophytes (ferns sensu lato), and two species of lycophytes. Wall preparations were obtained, extracted with 6M sodium hydroxide, and the extracts treated with a xyloglucan-specific endo-(1→4)-β-glucanase preparation. The oligosaccharides released were analysed by matrix-assisted laser-desorption ionisation time-of-flight mass spectrometry and by high-performance anion-exchange chromatography. The xyloglucan oligosaccharide profiles from the gymnosperm walls were similar to those from the walls of most eudicotyledons and non-commelinid monocotyledons, indicating that the xyloglucans were fucogalactoxyloglucans, containing the fucosylated units XXFG and XLFG. The xyloglucan oligosaccharide profiles for six of the monilophyte species were similar to those of the gymnosperms, indicating they were also fucogalactoxyloglucans. Phylogenetically, these monilophyte species were from both basal and more derived orders. However, the profiles for the other monilophyte species showed various significant differences, including additional oligosaccharides. In three of the species, these additional oligosaccharides contained arabinosyl residues which were most abundant in the profile of Equisetum hyemale. The two species of lycophytes examined, Selaginella kraussiana and Lycopodium cernuum, had quite different xyloglucan oligosaccharide profiles, but neither were fucogalactoxyloglucans. The S. kraussiana profile had abundant oligosaccharides containing arabinosyl residues. The L. cernuum profile indicated the xyloglucan had a very complex structure.

Evolution of milk oligosaccharides and lactose: a hypothesis

Mammalian milk or colostrum contains up to 10% of carbohydrate, of which free lactose usually constitutes more than 80%. Lactose is synthesized within lactating mammary glands from uridine diphosphate galactose (UDP-Gal) and glucose by a transgalactosylation catalysed by a complex of β4-galactosyltransferase and α-lactalbumin (α-LA). α-LA is believed to have evolved from C-type lysozyme. Mammalian milk or colostrum usually contains a variety of oligosaccharides in addition to free lactose. Each oligosaccharide has a lactose unit at its reducing end; this unit acts as a precursor that is essential for its biosynthesis. It is generally believed that milk oligosaccharides act as prebiotics and also as receptor analogues that act as anti-infection factors. We propose the following hypothesis. The proto-lacteal secretions of the primitive mammary glands of the common ancestor of mammals contained fat and protein including lysozyme, but no lactose or oligosaccharides because of the absence of α-LA. When α-LA first appeared as a result of its evolution from lysozyme, its content within the lactating mammary glands was low and lactose was therefore synthesized at a slow rate. Because of the presence of glycosyltransferases, almost all of the nascent lactose was utilized for the biosynthesis of oligosaccharides. The predominant saccharides in the proto-lacteal secretions or primitive milk produced by this common ancestor were therefore oligosaccharides rather than free lactose. Subsequent to this initial period, the oligosaccharides began to serve as anti-infection factors. They were then recruited as a significant energy source for the neonate, which was achieved by an increase in the synthesis of α-LA. This produced a concomitant increase in the concentration of lactose in the milk, and lactose therefore became an important energy source for most eutherians, whereas oligosaccharides continued to serve mainly as anti-microbial agents. Lactose, in addition, began to act as an osmoregulatory molecule, controlling the milk volume. Studies on the chemical structures of the milk oligosaccharides of a variety of mammalian species suggest that human milk or colostrum is unique in that oligosaccharides containing lacto-N-biose I (LNB) (Gal(β1 → 3)GlcNAc, type I) predominate over those containing N-acetyllactosamine (Gal(β1 → 4)GlcNAc, type II), whereas in other species only type II oligosaccharides are found or else they predominate over type I oligosaccharides. It can be hypothesized that this feature may have a selective advantage in that it may promote the growth of beneficial colonic bacteria, Bifidobacteria, in the human infant colon.

Mechanisms involved in enhancement of the expression and function of aggrecanases by hyaluronan oligosaccharides

Small hyaluronan (HA) oligosaccharides serve as competitive receptor antagonists to displace HA from the cell surface and induce cell signaling events. In articular chondrocytes, this cell signaling is mediated by the HA receptor CD44 and induces stimulation of genes involved in matrix degradation, such as matrix metalloproteinases (MMPs) as well as matrix repair genes including type II collagen, aggrecan, and HA synthase 2. The objective of this study was to determine changes in the expression and function of aggrecanases after disruption of chondrocyte CD44-HA interactions. Bovine articular chondrocytes or bovine cartilage tissue was pretreated with a variety of inhibitors of major signaling pathways prior to the addition of HA oligosaccharides. Changes in aggrecanase were monitored by real-time reverse transcription-polymerase chain reaction and Western blot analyses of ADAMTS-4, ADAMTS-5, and aggrecan proteolytic fragments. To test the interactions between ADAMTS-4 and membrane type 4 MMP (MT4-MMP), protein lysates purified from stimulated chondrocytes were subjected to coimmunoprecipitation. Disruption of chondrocyte CD44-HA interactions with HA oligosaccharides induced the transcription of ADAMTS-4 and ADAMTS-5 in a time- and dose-dependent manner. The association of glycosyl phosphatidylinositol-anchored MT4-MMP with ADAMTS-4 was also induced in articular chondrocytes by HA oligosaccharides. Inhibition of the NF-κB pathway blocked HA oligosaccharide-mediated stimulation of aggrecanases. Disruptive changes in chondrocyte-matrix interactions by HA oligosaccharides induce matrix degradation and elevate aggrecanases via the activation of the NF-κB signaling pathway.

Tolerance and Safety Evaluation in a Large Cohort of Healthy Infants Fed an Innovative Prebiotic Formula: A Randomized Controlled Trial

Backgroundthe addition of oligosaccharides to infant formula has been shown to mimic some of the beneficial effects of human milk. The aim of the study was to assess the tolerance and safety of a formula containing an innovative mixture of oligosaccharides in early infancy.Methodology/Principal Findingsthis study was performed as a multi-center, randomized, double-blind, placebo-controlled trial including healthy term infants. Infants were recruited before the age of 8 weeks, either having started with formula feeding or being fully breast-fed (breastfeeding group). Formula-fed infants were randomized to feeding with a regular formula containing a mixture of neutral oligosaccharides and pectin-derived acidic oligosaccharides (prebiotic formula group) or regular formula without oligosaccharides (control formula group). Growth, tolerance and adverse events were assessed at 8, 16, 24 and 52 weeks of age. The prebiotic and control groups showed similar mean weight, length and head circumference, skin fold thicknesses, arm circumference gains and stool frequency at each study point. As far as the anthropometric parameters are concerned, the prebiotic group and the control group did not attain the values shown by the breastfeeding group at any study point. The skin fold thicknesses assessed in the breastfeeding group at 8 weeks were strikingly larger than those in formula fed infants, whereas at 52 weeks were strikingly smaller. The stool consistency in the prebiotic group was softer than in the control group at 8, 16 and 24 weeks (p<0.001) and closer to that of the breastfeeding group. There was no difference in the incidence of adverse events between the two formula groups.Conclusionsour findings demonstrate the tolerability and the long term safety of a formula containing an innovative mixture of oligosaccharides in a large cohort of healthy infants.Trial Registration:drks-neu.uniklinik-freiburg.de DRKS 00000201

An additional oligosaccharide moiety in the HA of a pandemic influenza H1N1 candidate vaccine virus confers increased antigen yield in eggs

The H1N1 influenza pandemic in 2009 highlighted the need for the rapid generation of candidate vaccine viruses (CVVs) against an A/California/7/2009-like virus. The first available CVVs gave low protein yields in eggs but improved yields were achieved for second generation CVVs which contained amino acid substitutions compared to their precursor viruses. In this study, we investigated the basis for the increased virus protein yield of CVV NIBRG-121xp and whether the improved yield characteristics could be transferred between this virus and two other CVVs, NYMC X-179A and NYMC X-181. We generated variant viruses by reverse genetics to contain combinations of amino acid substitutions found in high yielding NIBRG-121xp and NYMC X-181. We found that the increase in total protein yield and functional HA yield of NIBRG-121xp in eggs is attributable to the single amino acid substitution K119N in the HA. We also found that the glycosylation of position 119 is essential for the improved virus protein yield in eggs. However, the K119N yield-enhancing effect was not transferable between viruses, nor was the N129D change found in high yielding NYMC X-181. However, position 119 may be a useful locus to monitor in future for viruses and CVVs with potentially high yield.

Saccharomyces cerevisiae chitin biosynthesis activation by N-acetylchitooses depends on size and structure of chito-oligosaccharides

BackgroundTo explore chitin synthesis initiation, the effect of addition of exogenous oligosaccharides on in vitro chitin synthesis was studied. Oligosaccharides of various natures and lengths were added to a chitin synthase assay performed on a Saccharomyces cerevisiae membrane fraction.FindingsN-acetylchito-tetra, -penta and -octaoses resulted in 11 to 25% [14C]-GlcNAc incorporation into [14C]-chitin, corresponding to an increase in the initial velocity. The activation appeared specific to N-acetylchitooses as it was not observed with oligosaccharides in other series, such as beta-(1,4), beta-(1,3) or alpha-(1,6) glucooligosaccharides.ConclusionsThe effect induced by the N-acetylchitooses was a saturable phenomenon and did not interfere with free GlcNAc and trypsin which are two known activators of yeast chitin synthase activity in vitro. The magnitude of the activation was dependent on both oligosaccharide concentration and oligosaccharide size.