[Pectic polysaccharides in the stem calli of the rowan tree Sorbus aucuparia L].

The water-soluble polysaccharide fractions SAcI and SAclI were isolated from rowan tree stem calli. The SAcI fraction was shown to contain compounds belonging to the arabinogalactan II group. The SAcII fraction, termed sorban, was found to contain pectic polysaccharides that make up the protopectin complex of the cell wall callusand that had a high content of galacturonic acid residues together with neutral sugars characteristic of rhamnogalacturonan I (RG-I) polysaccharides. Using methylation assays, some structural features of the ramified region of sorban were elucidated. The backbone of sorban was found to consist of 1,4-α-D-galacto-pyranosyluronic acid residues. The neutral sugars are represented by 1,6-linked galactopyranose and 1,5-linked arabinofuranose residues as the primary constituents, as well as 1,6-linked mannopyranose and 1,4-linked glucopyranose and xylopyranose residues. Callus growth was shown to be accompanied by nearly constant quantities of galacturonic acid and neutral sugar residues in sorban (fraction SAcII) from the rowan stem callus.

[Structural study of bergenan, a pectin from Bergenia crassifolia].

A pectin polysaccharide named bergenan was isolated from the freshly collected leaves of the leather bergenia Bergenia crassifolia by extraction with an aqueous solution of ammonium oxalate. The main component of its carbohydrate chain was shown to be the residues of D-galacturonic acid (about 80%). In addition, the polysaccharide contains residues of galactose, arabinose, and rhamnose; their total content is less than 15%. It was shown that the bergenan samples from bergenia leaves collected at different vegetation periods (from July to September) do not substantially differ either in monosaccharide composition or in the viscosity of aqueous solutions they form. The results of enzymatic hydrolysis by alpha-1,4-galacturonase (pectinase), partial acidic hydrolysis, NMR spectroscopy, and methylation with subsequent analysis of the results by GC-MS indicate that the bergenan macromolecule contains the regions of a linear alpha--1,4-D-galactopyranosyluronan and rhamnogalacturonan-I (RG-1). Galacturonan responds for a greater part of the macromolecule. A considerable amount of its constituent galacturonic acid residues are present as methyl esters. The side chains in RG-I are attached to the rhamnopyranose residues of the main carbohydrate chain by 1,4-link and are composed of the residues of terminal arabinofuranose and galactopyranose, 1,5-linked (-arabinofuranose, and 1,4-and 1,6-linked beta-galactopyranose. The branching points of the side chains of the RG-I molecule are 3,4- and 3,6-di-O-substituted galactose residues.

[Inhibition of neutrophil adhesion by pectic galacturonans].

The inhibition of the adhesion of neutrophils to fibronectin by the fragments of the main galacturonan chain of the following pectins was demonstrated: comaruman from the marsh cinquefoil Comarum polustre, bergenan from the Siberian tea Bergenia crassifolia, lemnan from the duckweed Lemna minor, zosteran from the seagrass Zostera marina, and citrus pectin. The parent pectins, except for comaruman, did not affect the cell adhesion. Galacturonans prepared from the starting pectins by acidic hydrolysis were shown to reduce the neutrophil adhesion stimulated by phorbol 12-myristate 13-acetate (1.625 microM) and dithiothreitol (0.5 mM) at a concentration of 50-200 microg/ml. The presence of carbohydrate chains with molecular masses higher than 300, from 100 to 300, and from 50 to 100 kDa in the galacturonan fractions was proved by membrane ultrafiltration.

Antiinflammatory activity of the pectic polysaccharide from Comarum palustre.

A pectic polysaccharide named comaruman (CP) was extracted from the aerial part of Comarum palustre with 0.7% aqueous ammonium oxalate and subsequent precipitation with ethanol. Oral administration of comaruman (5-100 mg/kg) was found to reduce a paw edema observed 24 h after injection of 2% formalin in mice. A fraction of comaruman (CP-H9) exhibited a similar antiinflammatory activity. Comaruman, CP deprived of lipid, CP purified by proteins and CP fractions obtained with acidic hydrolysis inhibit spontaneous and phorbol-12-myristate-13-acetate-activated adhesion of peritoneal leukocytes in vitro.

The structure of mytilan, a bioglycan-immunomodulator isolated from the mussel Crenomytilus grayanus.

Mytilan, isolated from the mantle of the mussel Crenomytilus grayanus and which possesses high immunomodulating activity, is a non-covalently linked complex of a polysaccharide (95%) and protein (5%) with lectin activities. Chromatography of mytilan on agarose separated the polysaccharide and protein constituents. The protein appeared to have the lectin properties. Methylation analysis, Smith degradation, digestion with alpha-amylase, and 13C-n.m.r. spectroscopy showed the polysaccharide to be a glycogen-like alpha-D-glucan.

[Isolation and characterization of polysaccharides from Tansy]. / Vydeleniia i obshchaia kharakteristika polisakharidov Pizhmy obyknovennoi.

Using extraction with 0.75% aqueous ammonium oxalate, the following polysaccharide fractions were isolated: tanacetans TVF, TVS, and TVR from floscules, sprouts, and roots, respectively, of Tanacetum vulgare L., spread throughout the European North of Russia. The sugar chain of tanacetan TVF consists of D-galacturonic acid (61.4%), arabinose (14.7%), galactose (10.2%), and rhamnose (3.7%) as the main constituents as well as xylose, glucose, mannose, apiose, and 2-O-methylxylose in trace amounts. Tanacetans TVS and TVR were shown to differ in the sugar quantitative composition. They contain 67 and 28% galacturonic acid, respectively. A partial acid hydrolysis of the tanacetan TVF gave a polysaccharide fragment TVF1, alpha-1,4-D-galacturonan (GalA 98.2%). Digestion with pectinase (alpha-1,4-D-polygalacturonase) resulted in fragment TVF3, containing residues of arabinose (27.1%) and galactose (17.3%). NMR spectroscopy allowed detection of the terminal residues of alpha-Araf and beta-Galp as well as of the residues of alpha-Araf substituted in 3,5- and 5-positions. Thus, tanacetan TVF was proved to be a pectic polysaccharide.

[Structure and physiological activity of lemnan, Lemna minor L. pectin]. / Strukturnoe issledovanie i fiziologicheskaia aktivnost' lemnana, pektina iz Lemna minor L..

A pectic polysaccharide, lemnan, was isolated from freshly collected duckweed Lemna minor L. Its sugar chain was shown to be mainly composed of the residues of D-galacturonic acid (64%), galactose, arabinose, xylose, and D-apiose, a branched chain sugar. The high content of D-apiose (25%) indicated that lemnan is an apiogalacturonan type pectin similar to zosteran, a pectic polysaccharide from a sea phanerogam of the Zosteraceae family. The results of partial acidic hydrolysis, pectinase digestion, and NMR studies of lemnan demonstrated that its macromolecule contains regions of the linear alpha-1,4-D-galacturonan and branched apiogalacturonan. The side chains of apiogalacturonan were found to be formed of single and 1,5-linked residues of D-apiofuranose attached to 2- and 3-positions of the D-galacturonic acid residues of the apiogalacturonan backbone. Lemnan was shown to exhibit an immunomodulatory effect by activating the system of phagocytosis.

[Isolation and structural study of polysaccharides from campion Silene vulgaris]. / Vydelenie i issledovanie stroeniia polisakharidov iz smolevki obyknovennoi Silene vulgaris.

Silenan SV, a pectic polysaccharide, was isolated from the aerial part of Silene vulgaris (Moench) Garke (Oberna behen (L.) Ikonn.), widespread through the European North of Russia. The polysaccharide was found to contain residues of galacturonic acid (63%), arabinose, galactose, and rhamnose as the main constituents. The results of a partial acidic hydrolysis, pectinase digestion, and NMR studies of silenan SV indicated that its molecule contains a linear alpha-1,4-D-galacturonan backbone and ramified regions. The core of the ramified regions is composed of residues of alpha-1,4-D-galacturonic acid along with 2-substituted alpha-rhamnopyranose residues. The NMR data showed that the silenan SV side chains are composed of the blocks built from the terminal alpha-1,5-linked arabinofuranose and beta-1,4-linked galactopyranose residues; these most likely are the side chains of rhamnogalacturonan, characteristic of other pectic polysaccharides. The nonreducing ends of these side chains contain alpha-arabinofuranose residues.

[The isolation, preliminary study of structure and physiological activity of water-soluble polysaccharides from squeezed berries of Snowball tree Viburnum opulus]. / Vydelenie i predvaritel'noe issledovanie stroeniia i fiziologicheskoi aktivnosti vodorastvorimykh polisakharidov iz shrota iagod Kaliny obyknovennoi Viburnum opulus.

Water-soluble polysaccharide fractions VO1-VO4 were isolated from the squeezed berries of snowball tree (Viburnum opulus) by successive extraction with water at various temperatures and pH and with aqueous solutions of ammonium oxalate. These fractions were purified by ion-exchange chromatography on DEAE cellulose, and the homogeneity of the purified polysaccharides was determined by gel filtration on Sephacryl S-500. Acidic polysaccharides close to pectins in their sugar composition were found in all the extracts (fractions VO1-1, VO2-1, VO3-2, and VO4-2). Residues of galacturonic acid, galactose, arabinose, and (to a lesser extent) rhamnose are their main constituents. Neutral polysaccharides composed mainly of galactose and mannose residues were additionally found in fractions extracted with acidified water (pH 4.0) and with aqueous ammonium oxalate solutions. Partial acidic hydrolysis and digestion with pectinase of acidic polysaccharides indicated that their carbohydrate backbone consists of alpha-1,4-linked residues of D-galacturonic acid. NMR spectra of acidic polysaccharides (fractions VO3-2 and VO3-3) confirmed this and demonstrated that their side oligosaccharide chains are composed of beta-1,4-linked galactopyranose residues and of terminal and 2,5- and 3,5-substituted residues of alpha-arabinofuranose at a Gal: Ara ratio of 3:1. Some polysaccharides from V. opulus were found to possess an immunostimulating activity: they enhance phagocytosis, in particular, the phagocytic index and the secretion of lysosomal enzymes with peritoneal macrophages. Calcium ions were found to be necessary for the appearance of the stimulating effect of acidic polysaccharides from V. opulus.

[Isolation and general characteristics of lectin from the ascidian Didemnum ternatum]. / Vydelenie i obshchaia kharakteristika lektina iz astsidin Didemnum ternatum.

DTL was isolated from the Didemnum ternatum colonial ascidian and purified by affinity chromatography on cross-linked ovalbumin followed by gel-filtration on Sephadex G-100. SDS-PAGE of the preparation showed a major intense band with a relative mol. wt. of ca. 27 kDa. In the presence of 2-mercaptoethanol, DTL gave rise to a single band of 10 kDa. The lectin isolated was found to agglutinate Ehrlich's carcinoma cells and trypsinized human A, B, O erythrocytes, the hemagglutination being inhibited by GlcNAc, chitobiose, desialylated glycoproteins. Comparison of the sugar moieties of the glycoproteins used as inhibitors led to suggestion that the specific sugar binding site contains, as a dominant sugar, a "bisecting" glucosamine attached through a beta-1,4-linkage to the beta-linked mannose residue of N-glycosidic chains of a complex or hybrid type. An absence of sialic acid residues linked with galactose residues appeared to be necessary for binding DTL with the complex-type sugar chains. DTL inhibits proliferation and reduces DNA biosynthesis ([3H]thymidine incorporation) in tumour cells (HeLa). Principal morphological differences were detected to occur between intact and DTL-treated HeLa cells.

[The immunomodulating action of mytilan in an experimental influenza infection]. / Immunomoduliruiushchee deistvie mitalana pri éksperimental'noi grippoznoi infektsii.

The materials characterizing the immunomodulating activity of mytilan, a bioglycane from an industrial object of Japanese sea, Crenomytilus grayanus, in experimental influenza infection are presented. The preparation protected up to 60% of mice intranasally inoculated with influenza virus, and significantly inhibited influenza virus reproduction. It induced interferon (possibly gamma IF) synthesis in low titres. The immunomodulating effect of mytilan consisted in stimulation of specific antibody genesis, enhancement of chemotaxis, increasing oxidation-reduction activity, enhancement of expression of Fc gamma-receptors by splenocytes and macrophages, enhancement of antibody-dependent spleen cell cytotoxicity, increased activity of natural killer cells, as well as increased complementary activity of blood serum and enhanced expression of C3 receptors by mouse spleen cells.

[Immunostimulating activity of the lipopolysaccharides of blue-green algae]. / Immunostimuliruiushchaia aktivnost' lipopolisakharidov sine-zelenykh vodoroslei.

The whole cells of blue-gree algae and lipopolysaccharides isolated from these cells were shown to stimulate the production of macro-(mainly) and microglobulin antibodies in rabbits. The macro- and microphage indices in rabbits increased significantly after the injection of LPS isolated from blue-green algae 24--48 hours before infecting the animals with a virulent Y. pseudotuberculosis strain. Besides, the inhibiting action of this strain on the migration of phagocytes to the site of infection was abolished immediately after the injection. The use of the indirect hemagglutination test allowed to prove the absence of close antigenic interrelations between blue-green algae and the following organisms: Spirulina platensis, Microcystis aeruginosa, Phormidium africanum and P. uncinatum.

[The lectin activity of mytilan, a bioglycan from mussels, and its effect on microbial adhesion to macroorganism cells]. / Lektinovaia aktivnost' mitilana, biokligana iz midii, i ee vliianie na adgeziiu mikrobov k kletkam makroorganizma.

One of the mechanisms of the protective action of a bioglycan isolated from the mantle of mussels of the species Crenomytalus grayanus in bacterial infections has been studied. The preparation under study has been found to possess the activity of galactose-specific lectin and considerably decreases the adhesive properties of Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. The results of this investigation open prospects for further study of this bioglycan as an agent preventing bacterial colonization in infectious diseases.

[Effect of mytilane, a bioglycan from the mussel Crenomytilus grayanus, on the course and outcome of experimental flu infection]. / Vliianie mitilana--bioglikana iz midii Crenomytilus grayanus na techenie i iskhod éksperimental'noi grippoznoi infektsii.

Materials characterizing the antiviral activity of Mytilane, manifested by the protection of 50-60% of mice infected with the lethal dose of influenza virus and by a decrease in the severity of pathological changes in the lungs of mice, are presented. The inhibiting activity of Mytilane with respect to the reproduction of influenza virus in vivo and in vitro under experimental conditions is demonstrated.

[Effect of mytilan, a polysaccharide from Crenomytilus grayanus, on the functional activity of peritoneal macrophages]. / Vliianie mitilana--polisakharida iz midii Crenomytilus grayanus--na funktsional'nuiu aktivnost' peritoneal'nykh makrofagov.

The studies showed that mytilan, a polysaccharide from Crenomytilus grayanus had a marked activating effect on macrophages evident from respective morphological changes and increased metabolic and functional activity of the macrophages. Exposure to mytilan resulted in an increase in the number of the macrophages and their size (optic microscopy), changes in their surface (scanning microscopy) and ultrastructural reconstruction of the cells (light microscopy). It was shown that subcutaneous and intraperitoneal administration of mytilan defined a decrease in the activity of 5'-nucleotidase in the macrophages of the peritoneal exudate from ++noninbred mice and mice CBA. Moreover, under the influence of mytilan the macrophage phagocytic activity against E. coli, S. aureus, P. aeruginosa and P. vulgaris increased in vivo and in vitro.

Chemical characterization and anti-inflammatory effect of rauvolfian, a pectic polysaccharide of Rauvolfia callus.

The pectic polysaccharide named rauvolfian RS was obtained from the dried callus of Rauvolfia serpentina L. by extraction with 0.7% aqueous ammonium oxalate. Crude rauvolfian RS was purified using membrane ultrafiltration to yield the purified rauvolfian RSP in addition to glucan as admixture from the callus, with molecular weights 300 and 100-300 kD, respectively. A peroral pretreatment of mice with the crude and purified samples of rauvolfian (RS and RSP) was found to decrease colonic macroscopic scores, the total area of damage, and tissue myeloperoxidase activity in colons as compared with a colitis group. RS and RSP were shown to stimulate production of mucus by colons of the colitis mice. RSP appeared to be an active constituent of the parent RS. The glucan failed to possess anti-inflammatory activity.

Structural studies of arabinogalactan and pectin from Silene vulgaris (M.) G. Callus.

Arabinogalactan and pectin (named silenan) were isolated from Silene vulgaris (M.) G. callus. Fractionation by ion-exchange chromatography on DEAE-cellulose and digestion with pectinase demonstrated that silenan from S. vulgaris callus (80% of D-galacturonic acid) and silenan from the aerial part of the campion S. vulgaris are similar: both pectins contain a high quantity of homogalacturonan segments. The NMR spectral data and mass spectrometry of the purified polysaccharide and its fragment obtained by Smith degradation confirmed that the core of the arabinogalactan consisted of the different segments of beta-1,3-D-galactopyranan. Some of the beta-galactopyranose residues of the backbone are branched at O-6. The side chains of the arabinogalactan were shown to contain residues of terminal and 3-O-substituted beta-galactopyranose, terminal alpha-arabinofuranose and alpha-rhamnopyranose, and 2-O-substituted alpha-rhamnopyranose. The alpha-rhamnopyranose residues in the sugar chain appeared to be 2-O-glycosylated by the beta-1,4-D-galactopyranosyl uronic acid residues.

Branching of the galacturonan backbone of comaruman, a pectin from the marsh cinquefoil Comarum palustre L.

Galacturonan, the main constituent of the backbone (core) of the comaruman macromolecule, a pectin from the marsh cinquefoil Comarum palustre L., was obtained on partial acid hydrolysis of the pectin. Using atomic force microscopy and methylation analysis of the galacturonan, the backbone of the comaruman macromolecule was shown to contain branches as side chains consisting of alpha-1,4-linked residues of D-galactopyranosyl uronic acid attached to the 2- and 3-positions of the galacturonic acid residues of the core, in addition to linear regions of alpha-1,4-D-galacturonan. A few side chains appear to attach to 2,3-positions of the D-galacturonic acid residues.