Water Extraction Kinetics of Bioactive Compounds of Fucus vesiculosus.

Brown macroalgae, particularly those from Fucus genus, are a rich and balanced source of bioactive nutrients and phytochemicals, such as dietary fibres (fucoidans, laminarins, and/or alginates), phlorotannins, and fucoxanthin, and some minerals, such as iodine, which have been demonstrated to possess numerous health-promoting properties. In fact, aqueous extracts of Fucus vesiculosus have been used as food supplements due to its rich content in bioactive compounds, though no study has been published on the optimization of this operation. Therefore, this study aimed to evaluate the impact of different extraction temperatures (25 °C, 50 °C, 75 °C, 100 °C, and 120 °C) and times (5 min, 1 h, 2 h, and 4 h) on the recovery of those bioactive compounds. The temperature was observed to positively influence the extraction of crude mass and of fucose polysaccharides only at 75 °C and above, and of iodine extraction at 50 °C and above. At these temperatures, time also showed to increase yields. Yields of crude extract, fucose, and iodine were successfully mathematically modelled with a power law, and its maximum yields were obtained at the highest temperature studied (120 °C) and longest extraction time (4 h). Iodine yield at these conditions provided extracts with relevant content to contribute to the recommended daily ingestion. Phlorotannins were significantly recovered at 120 °C though evidence of degradation was observed during time.

Natural vitamin B12 and fucose supplementation of green smoothies with edible algae and related quality changes during their shelf life.

BACKGROUND: Some algae are an excellent sources of vitamin B12, of special interest for vegetarian/vegan consumers, and of fucose to supplement fruit and vegetable beverages such as smoothies. Nevertheless, supplementation of smoothies with algae may lead to possible quality changes during smoothie shelf life that need to be studied. Therefore, the quality changes in fresh green smoothies supplemented (2.2%) with nine edible algae (sea lettuce, kombu, wakame, thongweed, dulse, Irish moss, nori, Spirulina and Chlorella) were studied throughout 24 days at 5 °C. RESULTS: The initial vitamin C content - 238.7-326.0 mg kg-1 fresh weight (FW) - of a 200 g portion of any of the smoothies ensured full coverage of its recommended daily intake, and still supplying 50-60% of the recommended intake after 7 days. Chlorella and Spirulina smoothies showed the highest vitamin B12 content (33.3 and 15.3 µg kg-1 FW, respectively), while brown algae showed fucose content of 141.1-571.3 mg kg-1 FW. These vitamin B12 and fucose contents were highly maintained during shelf life. CONCLUSION: The Spirulina supplementation of a 200 g smoothie portion ensured full coverage of the recommended vitamin B12 intake, with lower vitamin C degradation, during a shelf life of 17 days. Furthermore, thongweed and kombu are also considered as excellent fucose sources with similar shelf life. © 2017 Society of Chemical Industry.

Nutritional and bioactive compounds of commercialized algae powders used as food supplements.

The main nutritional/bioactive compounds (protein; aminoacids, AA; fucose; minerals; vitamins B12 and C; and total phenolic content, TPC) of nine commercial algae powders, used as food supplements, were studied. Undaria pinnatifida showed the highest protein/aminoacid contents (51.6/54.4 g 100 g-1). Among brown macroalgae, Himanthalia elongata showed the highest fucose content (26.3 g kg-1) followed by Laminaria ochroleuca (22.5 g kg-1). Mineral contents of 15-24% were observed in the algae, being particularly excellent sources of iodine (69.0-472.0 mg kg-1). Porphyra spp. and Palmaria palmata showed the highest vitamin B12 contents (667-674 µg kg-1). Vitamin C ranged among 490.4-711.8 mg kg-1. H. elongata showed the highest total phenolic content (14.0 g kg-1). In conclusion, the studied algae are excellent sources of protein, AA, minerals, vitamin C and some of them presented particularly high vitamin B12 and fucose contents, which may have a potential use as food supplements.

Chemical studies on the polysaccharides of Salicornia brachiata.

A group of 12 polysaccharide extracts were prepared from the tips, stem and roots of an Indian halophyte Salicornia brachiata Roxb. obtained by sequential extractions with cold water (CW), hot water (HW), aqueous ammonium oxalate (OX) and aqueous sodium hydroxide (ALK) solutions. Monosaccharide composition analysis revealed that all the polysaccharide extract samples consisted primarily of rhamnose, arabinose, mannose, galactose, glucose, whereas ribose and xylose were present only in some of the extracts. All the extracts exhibited low apparent viscosity (1.47-2.02 cP) and sulphate and contained no prominent toxic metal ions. Fucose was detected only in OX extract of the roots. These polysaccharides were found to be heterogeneous and highly branched (glycoside linkage analysis, size-exclusion chromatography, (13)C-NMR, FT-IR, circular dichroism and optical rotation data). Physico-chemical analyses of these polysaccharides including uronic acid, sulphate and protein contents were also carried out. This constitutes the first report on the profiling of Salicornia polysaccharides.

Rhamnogalacturonan II structure shows variation in the side chains monosaccharide composition and methylation status within and across different plant species.

A paradigm regarding rhamnogalacturonans II (RGII) is their strictly conserved structure within a given plant. We developed and employed a fast structural characterization method based on chromatography and mass spectrometry, allowing analysis of RGII side chains from microgram amounts of cell wall. We found that RGII structures are much more diverse than so far described. In chain A of wild-type plants, up to 45% of the l-fucose is substituted by l-galactose, a state that is seemingly uncorrelated with RGII dimerization capacity. This led us to completely reinvestigate RGII structures of the Arabidopsis thaliana fucose-deficient mutant mur1, which provided insights into RGII chain A biosynthesis, and suggested that chain A truncation, rather than l-fucose to l-galactose substitution, is responsible for the mur1 dwarf phenotype. Mass spectrometry data for chain A coupled with NMR analysis revealed a high degree of methyl esterification of its glucuronic acid, providing a plausible explanation for the puzzling RGII antibody recognition. The ß-galacturonic acid of chain A exhibits up to two methyl etherifications in an organ-specific manner. Combined with variation in the length of side chain B, this gives rise to a family of RGII structures instead of the unique structure described up to now. These findings pave the way for studies on the physiological roles of modulation of RGII composition.

Effects of Anethum graveolens L. on fertility in male rats.

OBJECTIVES: The effects of Anethum graveolens seed extract on fertility of male rats were investigated. METHODS: Male Wistar rats were divided into five groups according to the treatment they received during 42 days: control, low dose (0.5 g/kg) and high dose (5 g/kg) of aqueous extracts, and low dose (0.045 g/kg) and high dose (0.45 g/kg) of ethanol extracts of Anethum graveolens seed. Sperm count and motility and testosterone concentration were measured. Sections of the testes, epididymis, and seminal vesicles were stained with peroxidase-conjugated lectins of Ulex europaeus agglutinin, peanut agglutinin, Dolichos biflorus agglutinin, soy bean agglutinin and concanavalin A. The treated male rats were mated with females and the crown-rump lengths and weights of their newborn pups were measured. RESULTS: No significant differences in sperm count, sperm motility or testosterone concentration were observed in the experimental groups. However, female rats did not become pregnant after mating with rats given the high dose of the ethanol extract. The distribution of terminal sugars on the epithelial surface of the reproductive structures decreased in the experimental groups. CONCLUSION: Anethum graveolens extract decreased fertility rate by modifying some terminal sugars on the cell surface of male reproductive organs involved in sperm maturation, capacitation and oocyte recognition.

Effect of needle puncture and electro-acupuncture on mucociliary clearance in anesthetized quails.

BACKGROUND: Acupuncture therapy for obstructive respiratory diseases has been effectively used in clinical practice and the acupuncture points or acupoints of Zhongfu and Tiantu are commonly-used acupoints to treat patients with the diseases. Since the impaired mucociliary clearance is among the most important features of airway inflammation in most obstructive respiratory diseases, the effect of needle puncture and electro-acupuncture at the specific acupoints on tracheal mucociliary clearance was investigated in anesthetized quails. METHODS: Mucociliary transport velocity on tracheal mucosa was measured through observing the optimal pathway, and fucose and protein contents in tracheal lavages were determined with biochemical methods. In the therapeutic group, needle puncture or electro-acupuncture stimulation to the acupoints was applied without or with constant current output in 2 mA and at frequency of 100 Hz for 60 minutes. In the sham group, electro-acupuncture stimulation to Liangmen was applied. RESULTS: Our present experiments demonstrated that the electro-acupuncture stimulation to Zhongfu and Tiantu significantly increased tracheal mucociliary transport velocity and decreased the content of protein in the tracheal lavage, compared with the control group. Moreover, either needle puncture or electro-acupuncture stimulation to Zhongfu and Tiantu significantly reverted the human neutrophil elastase-induced decrease in tracheal mucociliary transport velocity and human neutrophil elastase -induced increase in the contents of fucose and protein in the tracheal lavage, compared with the control group. CONCLUSION: These results suggest that either needle puncture or electro-acupuncture stimulation to the effective acupoints significantly improves both airway mucociliary clearance and the airway surface liquid and that the improvements maybe ascribed to both the special function of the points and the substantial stimulation of electricity.

Bound sugars in hepatic glycoproteins from male rats during dietary citrus bioflavonoid and/or ascorbic acid supplementation.

The purpose of these experiments was to determine whether quantitative changes take place in the attached sugars of hepatic (postmitochondrial) glycoproteins isolated from rats fed a diet supplemented with citrus bioflavonoids (an equal mixture of rutin, naringin, and hesperidin) (B) and/or ascorbic acid (C) for 40 days and 90 days. Statistically significant increases in body weights (P < .05) were observed in the B-only groups and liver weights in the CB group (P < .01-.05) after 40 and 90 days of feeding the experimental diets, while liver weights were decreased in the B-only groups after 40 days of feeding (P < .05). In the acid-soluble glycoprotein fraction, statistically significant decreases were seen in bound hexoses and fucose (P < .05) in the CB group after 40 days, and in bound fucose only after 90 days (P < .05). In the acid-insoluble glycoprotein fraction, statistically significant changes were seen in bound hexoses and fucose (P < .05) in the CB group after 40 days, and in bound fucose only in the CB group after 90 days (P < .05). There is an apparent overall decrease in the sugar-rich acid-soluble glycoprotein fraction that is accentuated even further by combined CB supplementation. This decrease is more probable after 40 days than it is after 90 days of dietary supplementation. An adaptive phenomenon is suggested for maintaining the intracellular environment during periods of dietary citrus bioflavonoid and/or ascorbic acid supplementation.

Quantitative determination of saccharides in dietary glyconutritional products by anion-exchange liquid chromatography with integrated pulsed amperometric detection.

A new technique for the assay of carbohydrates is described in which separation and quantification of neutral saccharides, aminosaccharides, glycuronic acids, and disaccharides may be accomplished in less than 50 min of total run time. This method involves optimized anion-exchange liquid chromatography coupled with integrated pulse amperometric detection. Complex carbohydrates from various sources, including dietary supplements, were hydrolyzed in a dilute solution of trifluoroacetic acid, freeze-dried, and reconstituted in water containing 2-deoxygalactose as the internal standard. The solution was filtered and separated on CarboPac PA20 column. The eluted saccharides were detected by oxidation on a gold electrode with quadruple-pulsed integrated amperometry. The calibration plots for the saccharides were linear with an average correlation coefficient of 0.999. Method precision regarding peak retention time and resolution used in the peak identifications was verified. With this method, previously difficult-to-separate saccharides, such as galactosamine, glucosamine, and N-acetylglucosamine, were successfully resolved from the neutral saccharides rhamnose, arabinose, and galactose. Mannose was also resolved from xylose, and de-acetylation of aminosaccharides prior to separation was not necessary. This technique provides an accurate and efficient means to assay carbohydrates in dietary supplements, which new federal regulations will soon mandate.

[Studies on the constituents of polysaccharide and the content of saccharide from Mongolian medicine Hara Buri-16].

This article describes the extraction of dissolvable polysaccharide from Hara Buri-16, and using phenyl hydrate-sulfuric acid to determine the content of dissolvable polysaccharide. The average recovery is 101.1% and RSD is 0.94%. The components of water soluble polysaccharide were identified by gas chromatography as: fucose, arabinose, xylose, mannose, galactose and glucose in the molar ratio of 0.57:6.67:0.46:6.61:2.47:4.80 respectively.

[Purification and analysis of polysaccharides from the caudex of Undaria pinnatifida].

To isolate polysaccharides with hot water from the caudex of Undaria pinnatifida, and precipitate with ethanol. The protein in polysaccharides was removed by sevage way. DEAE-52 and Sephadex G-200 column chromatography were used to isolate and purification polysaccharides, three purified polysaccharides F2, F3 and F4 were obtained. It was identified that they were homogeneity. The ultraviolet spectrum showed there was no proteins and nucleic acids in F2, F3 and F4. Through thin-layer chromatography analysis, F2, F3 and F4 were mainly composed of Gal, Fuc, Man and Glu acid. F2 also contained Glu and Rha.

Structure of a heteroxylan of gum exudate of the palm Scheelea phalerata (uricuri).

The polysaccharide isolated from the gum exudate of palm Scheelea phalerata (SPN) was water-insoluble and composed of Fuc, Ara, Xyl, and uronic acid moieties in a 5:34:54:7 molar ratio: 12% of phenolics were also present. A soluble polysaccharide (SPNa) was obtained after alkaline treatment, which contained Fuc, Ara, Xyl and uronic acid in a 7:44:42:7 molar ratio, with only 2% phenolics. SPNa had an M(W) approximately 1.04 x 10(5) g mol(-1) and was almost monodisperse (M(W)/M(N) : 1.25 +/-0.22). It had a branched structure with side chains of 2-O-substituted Xylp (approximately 8%) and 3-O-substituted Araf (12%) units, and a large proportion of nonreducing end-units of Araf (15%), Fucp (10%), Xylp (4%), and Arap (6%). The (1 --> 4)-linked beta-Xylp main-chain units were 3-O- (9%), 2-O- (13%), and 2,3-di-O- (13%) substituted. Its (13)C NMR spectrum contained at least 9 C-1 signals, those at delta 108.6 and 107.7 arising from alpha-Araf units. Others were present at delta 175.4 from C-6 of alpha-GlcpA and delta 15.6 from C-6 of Fucp units. The main chain of SPNa was confirmed by analysis of a Smith-degraded polysaccharide (SPDS): methylation analysis provided a 2,3-Me(2)-Xyl (65%) derivative and its (13)C NMR spectrum showed five main signals typical of a (1 --> 4)-linked beta-Xylp units. Methylation analysis of a carboxy-reduced polysaccharide (SPN-CR) revealed a 2,3,4,6-Me(4)-Glc derivative (4%) arising from nonreducing end-units of GlcpA. Alpha-GlcpA-(1 --> 2)-alphabeta-Xy1p and alpha-GlcpA-(1 --> 2)-beta-Xylp-(1 --> 4)-alphabeta-Xylp were obtained via partial acid hydrolysis of SPN, showing the structure of side-chain substituents on O-2 of the main-chain units.

l-Galactose replaces l-fucose in the pectic polysaccharide rhamnogalacturonan II synthesized by the l-fucose-deficient mur1 Arabidopsis mutant.

Arabidopsis thaliana mur1 is a dwarf mutant with altered cell-wall properties, in which l-fucose is partially replaced by l-galactose in the xyloglucan and glycoproteins. We found that the mur1 mutation also affects the primary structure of the pectic polysaccharide rhamnogalacturonan II (RG-II). In mur1 RG-II a non-reducing terminal 2- O-methyl l-galactosyl residue and a 3,4-linked l-galactosyl residue replace the non-reducing terminal 2- O-methyl l-fucosyl residue and the 3,4-linked l-fucosyl residue, respectively, that are present in wild-type RG-II. Furthermore, we found that a terminal non-reducing l-galactosyl residue, rather than the previously reported d-galactosyl residue, is present on the 2- O-methyl xylose-containing side chain of RG-II in both wild type and mur1 plants. Approximately 95% of the RG-II from wild type and mur1 plants is solubilized as a high-molecular-weight (>100 kDa) complex, by treating walls with aqueous potassium phosphate. The molecular mass of RG-II in this complex was reduced to 5-10 kDa by treatment with endopolygalacturonase, providing additional evidence that RG-II is covalently linked to homogalacturonan. The results of this study provide additional information on the structure of RG-II and the role of this pectic polysaccharide in the plant cell wall.

Requirement of borate cross-linking of cell wall rhamnogalacturonan II for Arabidopsis growth.

Turgor-driven plant cell growth depends on wall structure. Two allelic l-fucose-deficient Arabidopsis thaliana mutants (mur1-1 and 1-2) are dwarfed and their rosette leaves do not grow normally. mur1 leaf cell walls contain normal amounts of the cell wall pectic polysaccharide rhamnogalacturonan II (RG-II), but only half exists as a borate cross-linked dimer. The altered structure of mur1 RG-II reduces the rate of formation and stability of this cross-link. Exogenous aqueous borate rescues the defect. The reduced cross-linking of RG-II in dwarf mur1 plants indicates that plant growth depends on wall pectic polysaccharide organization.

A hematopoietic cell L-selectin ligand that is distinct from PSGL-1 and displays N-glycan-dependent binding activity.

Human hematopoietic progenitor cells express L-selectin and also express PSGL-1, a ligand for all selectins. Using a shear-based adhesion assay, a hematopoietic cell L-selectin ligand (HCLL) that is expressed on the hematopoietic cell line KG1a and on normal human hematopoietic progenitors was previously identified. To characterize the structural biology of HCLL and to define its relationship to PSGL-1, the effects of chemical and enzymatic treatments on HCLL activity of KG1a cells and membrane preparations were analyzed. Protease digestions and chemical treatments of KG1a cells and membranes indicated that HCLL is an integral membrane glycoprotein. Glycosidase digestions of membrane protein preparations and metabolic treatments of KG1a cells with glycosylation processing modifiers revealed that L-selectin binding determinants on HCLL are sialofucosylated structures presented on complex-type N-glycans. Adhesion assays and biochemical studies showed that this glycoprotein is also expressed on circulating blasts in native acute leukemias. HCLL is distinguishable from PSGL-1: (1) KG1a cells sorted for PSGL-1 expression had equivalent HCLL activity; (2) anti-PSGL-1 blocking antibodies and proteases known to eliminate L-selectin binding to PSGL-1 had no effect on HCLL binding activity of KG1a cells; (3) blasts from native leukemias with low expression of PSGL-1 and CD34 display high HCLL activity; and (4) despite high level expression of PSGL-1, HCLL activity was absent on HL60 cells. These data provide first evidence of a naturally expressed membrane L-selectin ligand expressing binding determinant(s) on an N-linked glycoconjugate. This novel ligand may help mediate L-selectin-dependent cell-cell adhesive interactions within the cytoarchitecture of the bone marrow microenvironment. (Blood. 2000;96:2765-2774)

[Studies on polysaccharides from Holothuria leucospilota].

A powder was obtained from the dried body of Holothuria leucosilota (Brandt). According to IR and UV spectra, paper chromatography and the physical and chemical data, it is showed a sulfated mucopoly-saccharide consisted of galactosamine, glucouronic acid, fucose and sulfate with the molar ratio of 1:0.96:0.78:1.98 respectively. The specific rotation, intrinsic viscoslty and molecular weight of this polysaccharide were all presented.

[Extraction and purification of acidic mucopolysaccharide from Holothuria atra].

Mucopolyaccharide with molecular weight of 10253 Da was extracted and purified from fresh Holothuria atra Jaeger by means of enzymic and alkaline hydrolysis, potassium acetate and ethanol fractional precipitation. It was tested to be purified ingredient with agarose electrophoresis. The percentage content of galactosamine, glucuronic acid, fucose and sulfate in the mucopolysaccharide was 16.12%, 17.88%, 11.66% and 23.52% respectively.

Core alpha1,3-fucose is a key part of the epitope recognized by antibodies reacting against plant N-linked oligosaccharides and is present in a wide variety of plant extracts.

Carbohydrates have been suggested to account for some IgE cross-reactions between various plant, insect, and mollusk extracts, while some IgG antibodies have been successfully raised against plant glycoproteins. A rat monoclonal antibody raised against elderberry abscission tissue (YZ1/2.23) and rabbit polyclonal antiserum against horseradish peroxidase were screened for reactivity in enzyme-linked immunosorbent assay against a range of plant glycoproteins and extracts as well as neoglycoproteins, bee venom phospholipase, and several animal glycoproteins. Of the oligosaccharides tested, Man3XylFucGlcNAc2(MMXF3) derived from horseradish peroxidase was the most potent inhibitor of the reactivity of both YZ1/2.23 and anti-horseradish peroxidase to native horseradish peroxidase glycoprotein. The reactivity of YZ1/2. 23 and anti-horseradish peroxidase against Sophora japonica lectin was most inhibited by a neoglycoconjugate of bromelain glycopeptide cross-linked to bovine serum albumin, while the defucosylated form of this conjugate was inactive as an inhibitor. A wide range of plant extracts was found to react against YZ1/2.23 and anti-horseradish peroxidase, with particularly high reactivities recorded for grass pollen and nut extracts. All these reactivities were inhibitable with the bromelain glycopeptide/bovine serum albumin conjugate. Bee venom phospholipase and whole bee venom reacted weakly with YZ1/2.23 but more strongly with anti-horseradish peroxidase in a manner inhibitable with the bromelain glycopeptide/bovine serum albumin conjugate, while hemocyanin from Helix pomatia reacted poorly with YZ1/2.23 but did react with anti-horseradish peroxidase. It is concluded that the alpha1, 3-fucose residue linked to the chitobiose core of plant glycoproteins is the most important residue in the epitope recognized by the two antibodies studied, but that the polyclonal anti-horseradish peroxidase antiserum also contains antibody populations that recognize the xylose linked to the core mannose of many plant and gastropod N-linked oligosaccharides.

Wild type Rhizobium etli, a bean symbiont, produces acetyl-fucosylated, N-methylated, and carbamoylated nodulation factors.

Phaseolus vulgaris (common bean) can be nodulated by different Rhizobium species. A new species has been recently proposed: Rhizobium etli. Following transcriptional activation of the bacterial nodulation genes using naringenin or bean seed exudate, we have isolated, purified, and characterized R. etli extracellular nodulation factors. They are chitopentameric compounds that are N-methyl-N-vaccenoylated at their non-reducing end. At position 6 of the reducing N-acetyl-D-glucosamine, they are 4-O-acetyl-L-fucosylated. Minor compounds bear a carbamate group on the terminal non-reducing saccharidic residue.