Human milk oligosaccharides (HMO) are important immunoactive components found in breast milk. Scientific research proves that HMOs are significantly beneficial for infant health. 2'-fucosyllactose (2'-FL) is the major component of HMO, which obtained growing attentions from food industry. Besides, 3-fucosyllactose (3-FL) is another important fucosyllactose and it has a similar synthetic route comparing to 2'-FL. Thus, research of the two HMO components has interactive effects for each other. Recently, numerous publications are available for 2'-FL and 3-FL. The microbial cell factory is able to massively produce fucosyllactose via an efficient way, which will show considerable influences in dairy industry. In this paper, we review recent studies on 2'-FL and 3-FL, and discuss their prospects according to published literature and patents.
Female ; Humans ; Infant ; Milk, Human ; Oligosaccharides ; Trisaccharides

The interaction between viral HA (hemagglutinin) and oligosaccharide of the host plays an important role in the infection and transmission of avian and human flu viruses. Until now, this interaction has been classified by sialyl(alpha2-3) or sialyl(alpha2-6) linkage specificity of oligosaccharide moieties for avian or human virus, respectively. In the case of H5N1 and newly mutated flu viruses, classification based on the linkage type does not correlate with human infection and human-to-human transmission of these viruses. It is newly suggested that flu infection and transmission to humans require high affinity binding to the extended conformation with long length sialyl(alpha2-6)galactose containing oligosaccharides. On the other hand, the avian flu virus requires folded conformation with sialyl(alpha2-3) or short length sialyl(alpha2-6) containing trisaccharides. This suggests a potential future direction for the development of new species-specific antiviral drugs to prevent and treat pandemic flu.
Animals ; Antiviral Agents ; Classification ; Hand ; Humans ; Influenza in Birds ; Influenza, Human* ; Oligosaccharides* ; Oseltamivir* ; Pandemics ; Sensitivity and Specificity ; Trisaccharides

The aim of this study was to establish an assessment method for determining α-Gal (α-1, 3-galactosyle) epitopes contained in animal tissue or animal tissue-derived biological materials with ELISA inhibition assay. Firstly, a 96 well plate was coated with Gal α-1, 3-Gal/bovine serum albumin (BSA) as a solid phase antigen and meanwhile, the anti-α-Gal M86 was used to react with α-Gal antigens which contained in the test materials. Then, the residual antibodies (M86) in the supernatant of M86-Gal reaction mixture were measured using ELISA inhibition assay by the α-Gal coating plate. The inhibition curve of the ELISA inhibition assay, the R2 = 0.999, was well established. Checking using both α-Gal positive materials (rat liver tissues) and α-Gal negative materials (human placenta tissues) showed a good sensitivity and specificity. Based on the presently established method, the α-Gal expression profile of rat tissues, decellular animal tissue-derived biological materials and porcine dermal before and after decellular treatment were determined. The M86 ELISA inhibition assay method, which can quantitatively determine the α-Gal antigens contained in animal tissues or animal tissue-derived biomaterials, was refined. This M86 specific antibody based-ELISA inhibition assay established in the present study has good sensitivity and specificity, and could be a useful method for determining remnant α-1, 3Gal antigens in animal tissue-derived biomaterials.
Animals ; Antibodies ; Biocompatible Materials ; Enzyme-Linked Immunosorbent Assay ; methods ; Epitopes ; analysis ; Humans ; Rats ; Sensitivity and Specificity ; Serum Albumin, Bovine ; Trisaccharides ; analysis

BACKGROUND/AIMS: Icodextrin (glucose polymer) is metabolized by a-amylase to oligosaccharides such as maltose and maltotriose. The presence of these metabolites could have an effect on the enzymatic glucose measurement especially the glucose dehydrogenase pyrroloquinolinequinone (GDH-PQQ) based method. Patients treated with icodextrin are at risk for inaccurate blood glucose measurements. In this study we measured the blood glucose with different methods and analyzed the results to determine the test accuracy. METHODS: The blood glucose was measured, in seven outpatients and in seven inpatients using icodextrin, by the glucose hexokinase laboratory technique method as well as the GDH-PQQ method (Accu Chek Active)at the same time. To estimate an icodextrin residual effect, after discontinuing icodextin, the blood glucose was measured by the two methods after 48 hours in 4 inpatients. RESULTS: In seven outpatients the blood glucose was overestimated by the Accu Chek Active method (mean difference 68 mg/dL, p value 0.012). In seven inpatients the mean difference in the glucose was 56 mg/dL at 6am, 52 mg/dL at 11am, 52 mg/dL at 4pm, and 50 mg/dL at 9pm by the two different methods. In the four inpatients after changing their dialysate, the mean difference in the glucose was 58 mg/dL after 10 hours, 45 mg/dL after 24 hours, 24 mg/dL after 34 hours, and 26 mg/dL after 48 hours. CONCLUSION: Blood glucose was overestimated by the GDH-PQQ method and the inaccuracies were observed for more than 48 hours.
Blood Glucose ; Glucans ; Glucose ; Glucose 1-Dehydrogenase ; Hexokinase ; Humans ; Hypoglycemia ; Inpatients ; Maltose ; Oligosaccharides ; Outpatients ; Peritoneal Dialysis, Continuous Ambulatory ; Trisaccharides

BACKGROUND: Glutaraldehyde (GA) is a widely used cross-linking agent for improving mechanical properties and resistance to enzymatic degradation of collagenous tissue, but it has several drawbacks such as calcification and cytotoxicity. The aim of this study was to find the alternative effective cross-linking methods to GA. MATERIALS AND METHODS: Bovine pericardium was processed with GA with ethanol+octanol and glycine detoxification, and polyethylene glycol (PG) space filler, dimethyl 3,3'-dithiobispropionimidate (DTBP), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) treatment, and the physical fixation of ultraviolet irradiation were done. The biologic material properties of variously treated pericardial tissues were assessed by biochemical, mechanical and histological tests. Treated pericardial tissues were also implanted subcutaneously or intramuscularly into the rabbit for 10 weeks to assess the xenoreactive antibody response of immunoglobulin G and M, their anti-calcification effect. RESULTS: The biochemical and mechanical properties of EDC fixed pericardial tissues were comparable to the GA fixed tissue. The cytotoxicity was lowest in space filler treated GA fixed group. In rabbit subcutaneous or intramuscular implantation models, decellularization, space filler, EDC treatment group showed significantly lower calcium content than GA only and DTBP treatment group (p<0.05, analysis of variance). The titer of anti Galalpha1-3Galbeta1-4GlcNAc-R antibodies did not change in the postimplantation serial enzyme-linked immunosorbent assay. Hematoxylin and eosin and von Kossa staining showed that decellularization, space filler, EDC, and ultraviolet treatment had less inflammatory cell infiltration and calcium deposits. CONCLUSION: The decellularization process, PG filler, and EDC treatments are good alternative cross-linking methods compared to GA only fixation and primary amine of DTBP treatment for cardiovascular xenograft preservation in terms of the collagen cross-linking stability and in vivo anti-calcification effects.
Antibodies ; Antibody Formation ; Bioprosthesis ; Calcium ; Collagen ; Cyclohexanes ; Enzyme-Linked Immunosorbent Assay ; Eosine Yellowish-(YS) ; Glutaral ; Glycine ; Hematoxylin ; Imidoesters ; Immunoglobulin G ; Pericardium ; Polyethylene Glycols ; Transplantation, Heterologous ; Trisaccharides

BACKGROUND AND OBJECTIVES: Hyperacute rejection (HAR) is a major obstacle to successful xenotransplantation of vascularized organs. This study was conducted to observe the effect of hemolysis of perfused human whole blood on pig heart function, and determine the major risk factors for preservation of xenoperfused cardiac function using ex-vivo pig to human xenogeneic cardiac perfusion model. MATERIALS AND METHODS: Harvested pig hearts were perfused with normal human whole blood (group 1), two different types of pre-treated human whole blood (group 2: immunoglobulins were depleted by plasmapheresis, group 3: pre-treated with plasmapheresis, GAS914, cobra venom factor (CVF) and steroid), and normal porcine whole blood as control (group 4) for 3 hours. RESULTS: Duration of heart beat was significantly prolonged in group 2 and group 3. Histological examination showed widespread HAR features but was gradually delayed in groups 2 and 3 compared to group 1. The absolute levels of serum creatine kinase-MB and Troponin I increased gradually, and was lower in group 3. Serum hemoglobin levels were rapidly increased in groups 3 and 4, compared to group 1. Extracellular potassium level increased sharply from the beginning of blood perfusion in groups 1, 2 and 3, compared to group 4. CONCLUSION: Pretreatment of human whole blood, including immunoglobulin depletion, CVF and steroid reduced and delayed the destruction of pig myocardium by HAR. However, the increased extracellular potassium levels in groups 1, 2 and 3 reflected that these treatments could not prohibit myocardial injury by HAR.
Cobra Venoms ; Creatine ; Diphtheria Toxoid ; Extracorporeal Circulation ; Haemophilus Vaccines ; Heart ; Hemoglobins ; Hemolysis ; Humans ; Hyperkalemia ; Immunoglobulins ; Myocardium ; Perfusion ; Plasmapheresis ; Potassium ; Rejection (Psychology) ; Risk Factors ; Transplantation, Heterologous ; Trisaccharides ; Troponin I

OBJECTIVETo develop an HPLC method for the simultaneous quantitation of five constituents in Scrophularia ningpoensis.

METHODSamples were analyzed on an Agilent SB-C18 column(4.6 mm x 250 mm, 5 microm) eluted with acetonitrile and water containing 0.03% phosphate acid as mobile phases in a linear gradient mode. The flow rate was kept at 1.0 mL x min(-1), and the column temperature was set to 30 degrees C. The DAD detector wavelengths were 210, 280, 330 nm.

RESULTThe linear ranges were 50-400 mg x L(-1) for harpagide, 1-40 mg x L(-1) for harpagoside, 1-20 mg x L(-1) for cinnamic acid, 0.5-4.5 mg x L(-1) for acteoside,1-60 mg x L(-1) for angoroside C, respectively. The average recoveries of the five constituents were 100.8% (RSD 0.62%), 101.7% (RSD 0.32%), 98.8% (RSD 0.48%), 99.9% (RSD 1.4%), 99.2% (RSD 1.1%), respectively.

CONCLUSIONThrough the validation, the method was proved to be sensitive, accurate, repeatable, and can be used for quality control of the roots of S. ningpoensis.

Chromatography, High Pressure Liquid ; methods ; Cinnamates ; analysis ; Coumaric Acids ; analysis ; Glucosides ; analysis ; Glycosides ; analysis ; Iridoid Glycosides ; Phenols ; analysis ; Pyrans ; analysis ; Scrophularia ; chemistry ; Trisaccharides ; analysis

OBJECTIVETo make qualitative analysis on saccharide spots in thin layer chromatography (TLC) chromatogram of SI-WU-TANG extract C, which possesses blood-enrichment activity.

METHODTLC chromatogram of SI-WU-TANG extract C was obtained by using Automated Multiple Development (AMD) method. 4 major spots in the chromatogram were analyzed by off-line coupling TLC electrospray ionization mass spectrometry (ESI-MS) technique. Moreover, composition of monosaccharides in the fraction was analyzed by AMD technique.

RESULTMain constituents of substances from the 4 spots were monosaccharide, disaccharide, trisaccharide and tetrasaccharide respectively. Monosaccharide was mainly composed of fructose and glucose.

CONCLUSIONOff-line coupling TLC ESI-MS can simply and rapidly provide qualitative examination of saccharide spots in TLC chromatogram of Traditional Chinese Medicine. AMD method can make good separation of 8 frequently-observed monosaccharides in a regular 10 cm silica gel plate, the process of which was automated, AMD and off-line coupling TLC ESI-MS techniques show good value in saccharides analysis.

Angelica sinensis ; chemistry ; Chromatography, Thin Layer ; Disaccharides ; analysis ; Drugs, Chinese Herbal ; chemistry ; Fructose ; analysis ; Glucose ; analysis ; Ligusticum ; chemistry ; Monosaccharides ; analysis ; chemistry ; Paeonia ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rehmannia ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Trisaccharides ; analysis

The main commercial production of fructooligosaccharides (FOS) comes from enzymatic transformation using sucrose as substrate by microbial enzyme fructosyltransferase. A fructosyltransferase genomic DNA was isolated from Aspergillus niger QU10 by PCR. The nucleotide sequence showed a 1 941 bp size, and has been submitted to GenBank (KF699529). The cDNA of the fructosyltransferase, containing an open reading frame of 1 887 bp, was further cloned by RT-PCR. The fructosyltransferase gene from Aspergillus niger was functionally expressed both in Escherichia coli and Pichia pastoris GS 115. The highest activity value for the construction with the α-factor signal peptide reached 431 U/mL after 3 days of incubation. The recombinant enzyme is extensively glycosylated, and the active form is probably represented by a homodimer with an apparent molecular mass of 200 kDa as judged from mobility in seminative PAGE gels. The extracellular recombinant enzyme converted sucrose mostly to FOS, mainly 1-kestose and nystose, liberating glucose. FOS reached a maximal value and represented about 58% of total sugars present in the reaction mixture after 4 h reaction. The results suggest that the availability of recombinant Pichia pastoris as a new source of a FOS-producing enzyme might result of biotechnology interest for industrial application.
Aspergillus niger ; enzymology ; genetics ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Fungal Proteins ; genetics ; metabolism ; Glycosylation ; Hexosyltransferases ; genetics ; metabolism ; Molecular Sequence Data ; Molecular Weight ; Pichia ; Sucrose ; metabolism ; Trisaccharides ; metabolism

The saponins in different parts of Gynostemma pentaphyllum were analyzed via UPLC-Q-TOF-MS~E. A total of 46 saponins were identified, and the underground part had 26 saponins more than the aboveground part, most of which were trisaccharide saponins. The rat model of hyperlipidemia was established with high-fat diet. This study explored the lipid-lowering activity of total saponins in the underground part of G. pentaphyllum, so as to provide a theoretical basis for the comprehensive utilization of the underground part of G. pentaphyllum. A total of 99 healthy SD rats were randomly assigned into a blank group, a model group, a positive drug group, an aboveground total saponins group, and low-, medium-, and high-dose underground total saponins groups. Except the blank group, the other groups were fed with high-fat diet for 6 weeks. Then, the blood was collected from the orbital cavity to determine whether the modeling was successful according to the serum levels of total cholesterol(TC) and triglyceride(TG). After intragastric administration of the corresponding agents for 30 continuous days, the physical state of the rats were observed, and the body weight and liver specific gravity were measured. Furthermore, the levels of TC, TG, low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), alanine transaminase(ALT), aspartate transaminase(AST), bilirubin, and total bile acids in serum, as well as the levels of superoxide dismutase(SOD), malondialdehyde(MDA), peroxidase proliferator-activated receptor(PPAR-γ) in the liver tissue, were determined. The pathological changes of liver was observed via HE staining. The results showed that the aboveground total saponins and medium-and high-dose underground total saponins can treat hepatocyte steatosis, lower TC, TG, LDL-C, ALT, AST, total bilirubin, MDA, and PPAR-γ levels, and increase HDL-C and SOD levels in the model rats. The effect tended to be more obvious with the increase in dosage. Therefore, the total saponins in the underground part of G. pentaphyllum have good pharmacological effect of reducing blood lipid, which provides a theoretical basis for the comprehensive utilization of the underground part of G. pentaphyllum.
Alanine Transaminase/analysis* ; Animals ; Aspartate Aminotransferases/analysis* ; Bile Acids and Salts/blood* ; Bilirubin/blood* ; Cholesterol, LDL/blood* ; Diet, High-Fat/adverse effects* ; Gynostemma/chemistry* ; Hypolipidemic Agents/therapeutic use* ; Lipoproteins, HDL/blood* ; Liver/metabolism* ; Malondialdehyde/analysis* ; Peroxisome Proliferator-Activated Receptors/analysis* ; Rats ; Rats, Sprague-Dawley ; Saponins/therapeutic use* ; Superoxide Dismutase ; Triglycerides/blood* ; Trisaccharides/therapeutic use*