Polysaccharide Preparations Research Articles

The structure of an acidic trisaccharide component from a cell wall polysaccharide preparation of the green alga Tetraselmis striata Butcher

The structure of an acidic trisaccharide component from a cell wall polysaccharide preparation of the green alga Tetraselmis striata Butcher

Role of extracellular polysaccharide in the colonization of wheat (Triticum vulgare L.) roots by N2-fixing cyanobacteria

The characteristics of the mucilaginous sheaths of the cyanobacteria Nostoc 2S9B and Anabaena C5 and their role in the formation of associations with the roots of wheat plants grown in liquid culture have been assessed. Light and scanning electron microscopy revealed that the filaments of Nostoc 2S9B that formed a tight association with the root surface were contained in a firm mucilaginous shell. In contrast, filaments of Anabaena C5 formed a loose association and were easily detached from the mucilage that had a sheet-like appearance and tended to disintegrate as the culture aged. Similarly, there was a tight attachment of the isolated polysaccharide from Nostoc 2S9B to the root surface and a loose attachment of the Anabaena C5 polysaccharide. When the crude polysaccharide from Nostoc 2S9B was freed from proteins by phenol or pronase treatment, its ability to adhere to the root surface was lost or considerably reduced, suggesting that a protein component contributes to the tight attachment of Nostoc 2S9B. The crude polysaccharide preparation from Nostoc 2S9B contained 2.8% (w/w) protein while that from Anabaena C5 was only 0.6% (w/w) protein. The purified exopolysaccharide from Nostoc 2S9B contained three neutral sugars and glucuronic acid, whereas fucose and a uronic acid were the main components of that from Anabaena C5. Washing the roots or treating them with different sugars did not alter the ability of Nostoc 2S9B to colonize the root surface, indicating that cyanobacterial attachment may not be specific.

The elevation of plasma soluble tumor necrosis factor receptor levels by TNF induction therapy for patients with malignancy.

Soluble tumor necrosis factor receptor (sTNF-R) is known to inhibit patient immunity via specific binding with the TNF molecule. To examine the possible involvement of sTNF-R in cancer immunotherapy, the plasma levels of sTNF-R of both 55 kDa and 75 kDa origins were estimated when TNF was induced in patients with malignancy using both a polysaccharide preparation (Lentinan) and a streptococcal preparation (OK-432). The pretreatment plasma levels of the 55 kDa and 75 kDa sTNF-R were 1.04 +/- 0.53 and 1.06 +/- 0.34 ng/ml (mean +/- SE), respectively. The plasma levels of TNF were undetectable before treatment. The plasma sTNF-R levels peaked 2 h after the administration of OK-432 and followed the same pattern as the TNF levels in plasma. Both TNF and sTNF-R nearly returned to pretreatment levels at 16 h after the induction of TNF. The peak plasma levels of the 55 kDa and 75 kDa sTNF-R were 2.46 +/- 0.95 and 3.03 +/- 0.88 ng/ml, respectively, but they did not correlate with the plasma TNF levels. When peripheral white blood cells were cultured with the addition of lipopolysaccharide in vitro, an elevation of the 72 kDa sTNF-R was detected. Thus, the plasma source of this soluble receptor can at least be partly attributed to the white blood cells. However, the 55 kDa sTNF-R showed little increase in the cultures, and its source remains unknown. We should therefore be aware of the elevation of plasma sTNF-R levels by the induction therapy of TNF for patients with malignancies because of the immunosuppressive effect of sTNF-R.

Structural Studies on a Cell Wall Polysaccharide Preparation of Lactococcus Lactis Subspecies Cremoris H414

Abstract A cell wall polysaccharide preparation of Lactococcus lactis subsp. cremoris H414 was isolated by nitrous acid extraction, and purified by gel filtration and anion-exchange chromatography. Oligosaccharide fragments, obtained by partial acid hydrolysis or alkaline hydrolysis, were studied by means of monosaccharide analysis, methylation analysis, FAB-MS, EI-MS, and 1H NMR spectroscopy. In addition 1H, 13C, and 31P NMR spectroscopy, including two-dimensional homo- and heteronuclear measurements, were performed on the native polysaccharide. Combination of the various results demonstrated that in the polysaccharide preparation two repeating units with the structures [→2)-α-l-Rhap-(1→3)-α-d-Galp-(1→]n and [→6)-[α-d-Galp-(1→4)]-α-d-GalpNAc-(1→3)-β-d-GalpNAc-(1→1)-Ribitol-(4-phosphate→]m, respectively, can be recognized. So far, a covalent linkage between both polymers could not be confirmed nor be excluded.

Antitumor Effect of Intratumoral Administration of Biological Response Modifiers: Induction of Immunosuppressive Acidic Protein, a Type of α1‐Acid Glycoprotein, in Mice

The antitumor effects of biological response modifiers (BRMs) in an experimental mouse model, the “double grafted tumor system” were analyzed. Male BALB/c mice received simultaneous inoculations of Metn‐A fibrosarcoma cells on the right flank (106 cells) and left flank (2 × 105 cells) on day 0, and BRMs were injected intratumorally into the right tumor on days 3, 4 and 5. PSK (a protein‐bound polysaccharide preparation), interleuldn‐1 (IL‐1) and cepharanthin (R) cured not only the right, but also the left, non‐treated tumor in a double grafted tumor system. OK‐432 (a Streptococcus preparation) and BCG and tumor necrosis factor (TNF) cured the right tumor and inhibited the growth of the left tumor. Lentinan (a polysaccharide preparation) and IL‐6 inhibited neither the right nor the left tumor. Immunosuppressive acidic protein (IAP) in serum was increased transiently soon after intradermal injection of PSK, CR, OK‐432 and TNF in BALB/c mice. Lentinan, however, did not induce IAP. IAP in serum was gradually increased after intradermal inoculation of Meth‐A tumor in BALB/c mice. The biochemical difference between PSK‐induced IAP (early, inflammatory IAP) and Meth‐A‐induced IAP (late, tumor‐induced IAP) was investigated by crossed affinity immunoelectrophoresis with concanavalin A. IAP of murine serum was separated into 4 peaks. IAP in normal mouse was rich in high‐mannose type sugar chain (Peak 3) and contained no hybrid‐type sugar chain (Peak 4), which was present in inflammatory and tumor‐induced IAP. Inflammatory IAP was rich in biantennary sugar chain (Peak 2) and tumor‐induced IAP was rich in tri‐tetraantennary sugar chain (Peak 1).

Hypocholesterolaemic non-starch polysaccharide from sugar beet

In four feeding trials with male Wistar rats the influences on serum cholesterol of several preparations of non-starch polysaccharide (NSP) from sugar beet were determined. These include a commercial product called Beta-fibre, hot water insoluble and soluble sugar-beet NSP (both pectic and hemicellulosic fractions) and a commercially developed pectic-hemicellulosic sugar-beet NSP called Fibre N. Animals were fed basal diets, both low and high in cholesterol, and intake was limited to circa 80% of ad libitum. The influence of ∼80 g NSP per kg basal diet administered for 28 days was compared with a cellulose control. In one feeding trial the basal low-cholesterol diet was made high in butter fat (230 g per kg). As with ad libitum fed rats reported in the literature, the serum cholesterol concentration did not increase on increasing the saturated fat intake, but doubled when a mixture of cholesterol and cholic acid was added to the diet. Soluble NSP from sugar beet, but not insoluble NSP, was found to be mil...

Effect of PSK, a protein‐bound polysaccharide preparation, on liver tumors of Syrian hamsters induced by Thorotrast injection

The contrast medium Thorotrast, an agent well known to be carcinogenic, was injected into 400 congeneic Syrian hamsters. The resulting incidence of malignant hepatic tumors such as cholangiocarcinoma, hepatocellular carcinoma and hemangiosarcoma, was significantly higher in the male experimental group than in the control group, and the 50% survival period in the male group was shortened by about 100 days (P < 0.01). However administration of the antitumor drug PSK (Polysaccharide Kureha), a protein bound-polysaccharide extracted from basidiomycete fungi, prevented this carcinogenic effect. The incidence of malignant hepatic tumors in the experimental group was 22.5% compared with 2.8% in the control group (P < 0.01) and 10.5% in the PSK-treated group (P < 0.01). PSK also increased the 50% survival period by 61 days (P < 0.01).

An Efficient Method for the Syntheses of Novel Amphiphilic Polysaccharides by Regio- and Thermoselective Modifications of Chitosan

Abstract The first example of an unique and efficient method for the preparation of amphiphilic polysaccharides having self-assembling nature is described. Regio- and thermospecific chemical manipulations of a standardized intermediate derived from chitosan markedly facilitated the synthetic procedures of novel types of polysaccharide architectures. Artificial glycolipid-type polymers showed excellent properties to form stable monolayer membranes at air/water interface.

Molecular size analysis of capsular polysaccharide preparations from Streptococcus pneumoniae

Purified capsular polysaccharide preparations from Streptococcus pneumoniae serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F were analyzed by high performance size exclusion chromatograpy (HPSEC) with multi-angle laser light scattering (MALLS), specific viscosity (SV), and refractive index (RI) detection to determine the molecular size and molar mass of each of the pneumococcal (Pn) polysaccharides. The M W′s of the polysaccharides ranged from a low of 606 kg/mol for Pn4 to a high of 1145 kg/mol for Pn9V, and the z-average radii of gyration ranged from 59 nm for Pn14 to 72 nm for Pn18C. Estimations of molar mass of the highly anionic polysaccharides (all but Pn14) by the universal calibration approach were unsuccessful, resulting in a 27–53% overestimate of the M W′s, though application of Mark-Houwink-Sakurada coefficients calculated from the HPSEC-MALLS/SV/RI data resulted in estimates of M W that were in agreement with the MALLS estimates for all but the Pn4 preparation. These results emphasize the need for direct measurement of both molecular size and intrinsic viscosity distributions for definitive characterization of the molar mass, hydrodynamic volume, rigidity, and drainage of complex biological polymers such as the pneumococcal polysacchrides.

Malic Acid-Containing Polysaccharide Produced by the Fungus Aureobasidium pullulans of Marine Origin.

Aureobasidium pullulans designated AL-2 strain, isolated from the intestine of the marine fish Hexagrammos otakii, produces an extracellular acidic polysaccharide containing malic acid. The polysaccharide was extracted with 1% phenol solution and purified by the addition of ethanol followed by precipitation with Cetavoln and DEAE-cellulose column chromatography. This polysaccharide preparation, which was homogeneous in both ultracentrifugal and electrophoretic analyses, was composed of glucose and malic acid at a molar ratio of 1:1.

The Cell Wall Galactan of Catenella nipae Zanardini from Southern Australia

The cell wall polysaccharide of Catenella nipae Zanardini collected from southern Australia was extracted and analysed by chemical and physical methods. Galactose and 3,6-anhydrogalactose were the major monosaccharide constituents of this preparation in a molar proportion of 6: 4, with minor amounts of xylose also present. No mono-O-methylgalactosyl residues were detected. The high sulphate ester content (ca. 30% as SO3Na) and gelling in the presence of Ca2+ are indicative of a carrageenan type of polysaccharide. Infrared and C-13-NMR spectroscopy established that the dominant component of the extracted polysaccharide preparation is iota (iota)-carrageenan.

The protective activity of immunostimulants against Listeria monocytogenes infection in mice

The function of peritoneal macrophages induced by intraperitoneal (i.p.) injection of attenuated Streptococcus pyogenes (OK-432), Bacillus Calmette Guérin (BCG), protein-bound polysaccharide preparation isolated from Coriolus vesicolor (PSK) or Lactobacillus casei was examined. The PMA-triggered respiratory burst (production of O2- and H2O2) of macrophages induced by OK-432, BCG or Lac. casei was greater than that of resident or thioglycollate-stimulated macrophages and was similar to that of BCG-activated macrophages. PSK failed to enhance the production of O2- or H2O2 by macrophages. Alkaline phosphodiesterase (APD) activity was reduced in macrophages induced by OK-432, BCG or Lac. casei injection and in BCG-activated macrophages. The APD activity of macrophages obtained 7 and 13 days after i.p. injection of PSK was elevated, as with thioglycollate-stimulated macrophages. Listericidal activity in vitro was enhanced in macrophages obtained in 13 and 7 days, but suppressed in macrophages obtained 2 days after OK-432, BCG or Lac. casei injection. Lac. casei administered either 2 or 13 days before infection with Listeria monocytogenes was protective but OK-432, BCG (0.1 mg) and PSK were not, even though they were able to stimulate macrophage function.

A Glutamic Acid-Containing Mucopolysaccharide from a Marine Pseudomonas.

A marine strain of Pseudomonas (No. 42), which was originally isolated from seawater, pro-duced an acidic mucopolysaccharide containing glutamic acid when grown on seawater agar medium containing sucrose. The polysaccharide was extracted with 1% phenol solution and purified by the addition of ethanol, followed by precipitation with Cetavlon and DEAE-cellulose chromatography. This polysaccharide preparation was homogeneous in both ultracentrifugal and electrophoretic analyses. This polysaccharide was composed of glucosamine, glucuronic acid, and glutamic acid.

Isolation and Characterization of a Sulfated Polysaccharide Produced by a Marine Bacterium.

A marine bacterium designated MIL-1, which was originally isolated from the green algae Codium fragile produced a sulfated polysaccharide when grown in a seawater medium containing sucrose. Isolation of this polysaccharide was accomplished by precipitation with ethanol and Cetavlon, followed by DEAE-cellulose column chromatography. This polysaccharide prepar-ation which was homogeneous in both ultracentrifugal and electrophoretic analyses was composed of galactose, rhamnose, and sulfate at a molar ratio of 1:1:1, together with O-acetyl groups.

Simultaneous Production of Muco- and Sulfated Polysaccharides by Marine Pseudomonas.

A marine Pseudomonas species designated WAK-1 strain which was originally isolated from the brown seaweed Undaria pinnatifida produces an acidic muco (A-1)- and a sulfated (B-1) poly-saccharides when grown in a seawater medium containing sucrose. Isolation and purification of these polysaccharides have been accomplished by precipitation with ethanol and Cetavlon, fol-lowed by DEAE-cellulose column chromatography. These polysaccharide preparations were homogeneous in both ultracentrifugal and electrophoretic analyses. A-1 was composed of gala-tosamine, glucuronic acid, and pyruvate at a molar ratio of 3:1:1, while B-1 was composed of galactose, glucose, and sulfate at a molar ratios of 2:1:2.

Effect of lymphokine-activated killer cells on human retinoblastoma cells (Y-79) in vitro: Enhancement of the activity by a polysaccharide preparation, krestin

Peripheral blood mononuclear cells (PBMC) cultured in a medium containing interleukin 2 (IL 2) develop the ability to kill fresh tumor cells. This function has been termed lymphokine activated killing(LAK). Recently, cord LAK cell activity was demonstrated to be equally as cytotoxic against similar in vitro targets as adult (peripheral) LAK cells. We investigated the future therapeutic use of LAK adoptive immunotherapy by examining LAK in vitro cytotoxicity from both cord and peripheral blood mononuclear cells against pediatric malignant tumor cell lines Y-79 (retinoblastoma). Cord LAK cells show higher levels of cytotoxicity toward Y-79 targets than do adult LAK cells. Attempts to enhance the rIL 2-induced LAK activity by addition of rIFN-γ or PSK (krestin) were successful. Furthermore, we found that PSK has a function to enhance rIL 2-induced IFN-γ and TNF-α production. These findings suggest that combined administration of cord LAK cells and PSK may account for the improvement of advanced retinoblastoma in the neonatal period.

Isolation and Characterization of a Fucosamine-Containing Polysaccharide from a Marine Strain of Pseudomonas.

A marine strain of Pseudomonas (HA-318) produces a polysaccharide with a rare component, fucosamine. The polysaccharide was extracted with 1% phenol solution and purified by addition of ethanol followed by precipitation with Cetavlon and DEAE-cellulose column chromatography. This polysaccharide preparation, which was homogeneous in both ultracentrifugal and electro-phoretic analyses, was composed of galacturonic acid, galactose, fucosamine, and pyruvate in the molar ratios of 3:1:1:1, together with N-acetyl groups.

Preparation of bacterial polysaccharide of hibridized enzyme susceptibility.

Acetobacter xylinum strains produced bacterial cellulose incorporated N-acetyl-glucosamine (GlcNAc) in molecule (N-AcGBC) in medium containing GlcNAc. The profiles of the N-AcGBC and normal bacterial cellulose (BC) microfibrils treated with cellulase or lysozyme were observed by scanning electron microscopy. After the treatment with lysozyme, the morphological changes of the N-AcGBC microfibrils of pellicle or homogenates were found. Cellulase did not make any difference between BC and N-AcGBC.

Polysaccharide Fraction from Higher Plants which Strongly Interacts with the Cytosolic Phosphorylase Isozyme

From leaves of Spinacia oleracea L. or from Pisum sativum L. and from cotyledons of germinating pea seeds a high molecular weight polysaccharide fraction was isolated. The apparent size of the fraction, as determined by gel filtration, was similar to that of dextran blue. Following acid hydrolysis the monomer content of the polysaccharide preparation was studied using high pressure liquid and thin layer chromatography. Glucose, galactose, arabinose, and ribose were the main monosaccharide compounds. The native polysaccharide preparation interacted strongly with the cytosolic isozyme of phosphorylase (EC 2.4.1.1). Interaction with the plastidic phosphorylase isozyme(s) was by far weaker. Interaction with the cytosolic isozyme was demonstrated by affinity electrophoresis, kinetic measurements, and by (14)C-labeling experiments in which the glucosyl transfer from [(14)C]glucose 1-phosphate to the polysaccharide preparation was monitored.

Gastroenteritis in Suckling Mice Caused by Human Rotavirus Can Be Prevented with Egg Yolk Immunoglobulin (IgY) and Treated with a Protein‐Bound Polysaccharide Preparation (PSK)

Oral inoculation of human rotavirus MO strain (serotype 3) into 5-day-old BALB/c mice cause gastroenteritis characterized by diarrhea. Clinical symptoms, histopathological changes in the small intestine, and the detection of rotavirus antigen in enterocytes were all characteristic of rotavirus-induced gastroenteritis. Using this small animal model, passive protection of suckling mice against human rotavirus infection was achieved with the use of immunoglobulin (IgY) from the yolks of eggs of rotavirus-immunized hens. When IgY against a rotavirus strain homotypic to the challenge virus (MO strain) was administered in the mice, complete protection against rotavirus infection was achieved. On the other hand, with oral administration of IgY against a heterotypic strain (serotype 1, Wa strain), a lower protective effect was nevertheless obtained. The four different strains of human rotavirus (Wa, KUN, MO, and ST3) were inactivated in vitro by treatment with PSK, a protein-bound polysaccharide preparation, in a dose-dependent manner. Oral administration of 2.5 mg of PSK caused a therapeutic effect on experimentally MO-infected suckling mice. The antiviral effect of PSK was indicated by the reduction of the duration of diarrhea.