[Efficiency of a new synbiotic drink in treatment of chronic diseases of gastrointestinal tract and concomitant dysbacteriosis].

The efficacy of a novel synbiotic drink in the complex therapy of patients withchronic diseases of the gastrointestinal tract and concominant intestinal dysbacteriosis was investigated in a randomized trial. The synbiotic drink contains a probiotic strain of bifidobacteria and Fucus evanescens polysaccharides with prebiotic activity and broad spectrum of the biological action on the patients. The use of the synbiotic drink provided more evident reduction of the clinical symptomes, more efficient recovery of the intestinal microflora and higher percentage of the patients cure vs. the routine therapy and the therapy with inclusion of sour milk bifidobacterin.

[Influence of intravenous injection of fucoidan from brown seaweed Fucus evanescens by plasma rabbits anticoagulant activity and neutralisation by sulphate protamin of fucoidans antithrombin activity in vitro].

With fucoidan from Fucus evanescens dose increase from 1 to 5 mg/kg plasma coagulation time in test A(see symbol)TB increases. Sulphate protamin in final concentration 0.67-1.35 mkg/ml will neutralise antithrombin activity of fucoidans from brown seaweed Fucus evanescens and Laminaria cichorioides. The gravimetrichesky relation for the investigated samples makes an antidot/anticoagulant 1.

Correlation between influence of polysaccharides on hydrolase activity and their antiviral effect in tobacco leaves.

The activities of hydrolases (acid phosphatase, RNase, and proteases) in healthy and tobacco mosaic virus-infected leaves of Nicotiana tabacum L. var. Samsun, both untreated and treated with polysaccharides (PS) (1,3;1,6-ß-D-glucan, fucoidan, and κ/ß-carrageenan), were determined. The PS lead to substantial increase in the hydrolase level. The percentage of viral particles undergoing destructive change also increases in leaves treated with PS 24 h before infection. We suppose that the PS-mediated hydrolase activation promotes intracellular destruction of the viral particles and, thus, comprises one of the PS-induced protective mechanisms limiting intracellular viral accumulation.

[Production of cytokines by murine bone marrow dendritic cells in vitro mediated by sulfated polysaccharides obtained from sea brown algae].

AIM: To assess in vitro cytokine production by murine bone marrow dendritic cells (DC) matured under the effect of sulfated polysaccharides--fucoidanes from sea brown algae Laminaria cichorioides and Laminaria japonica. MATERIALS AND METHODS: CBA line mice were used to obtain bone marrow origin precursors of DC. Isolation and study of chemical composition and structure of fucoidanes were performed using modern research methods. Expression of surface markers was determined by flow cytometry (FACS-analysis) using monoclonal antibodies to respective antigens. Levels of cytokine production were measured by t-ELISA using kits manufactured by Biosource (Belgium). RESULTS: I was determined that fucoidans induce maturation of DC that was evident by expression of terminal differentiation marker CD83, activation marker CD38, enhanced expression of costimulating CD86, antigen-presenting MHC II and adhesive CD11c molecules. Fucoidanes stimulate DC to produce proinflammatory (TNF-alpha, IL-6, IL-1beta) and regulatory (IL-12) cytokines. Fucoidanes enhance expression of TLR-2 and TLR-4 but do not influence on expression of TLR-9. CONCLUSION: It was shown that fucoidanes from sea brown algae L. cichorioides and L. japonica activate innate immunity system that is evident by enhanced expression of surface molecules associated with DC maturation and increased production of proinflammatory and regulatory cytokines by DC. Enhanced expression of TLR-2 and TLR-4 allows to suppose that studied fucoidanes could have anti-infective effect in vivo.

[Effect of fucoidan on the ultrastructure of mesophyll cells of Datura stramonium L. and accumulation of potato virus X in them].

Influence of fucoidan from brown alga Fucus evanescens C. Ag. on the development of infection induced by potato virus X (PVX) in Datura stramonium leaves was studied. It as been shown that 24 h after the treatment of the leaves with fucoidan and following infection of them with PVX the accumulation of virus particles in infected cells during early infection period was substantially less than that in untreated control. Using ultrastructure-morphometric analysis, it has been established that fucoidan treatment increases at protein-synthesizing capability of cells (nucleolus dimension, amount of mitochondria and rough endoplasmic reticulum membranes become increased). At the same time, the fucoidan treatment causes some activation of lytic compartment which leads to destruction of virus particles and, therefore, might be considered as one of fucoidan-dependent protective mechanisms limiting virus accumulation in cells. Fucoidan stimulation of the formation of PVX-specific laminated structures capable of virus particles binding is possibly another induced antiviral cell mechanism, preventing from virus reproduction and transposition.

[Inhibiting effects of fucoidans on Hantaan virus adsorption on a model of peritoneal macrophages in vitro].

A model of peritoneal macrophages was used to study the effect of fucoidans from brown seaweeds on the adsorption of Hantaan virus. Fucoidans were found to have antiviral activity, but to differ in the inhibition of hantavirus adsorption, which was associated with their structural features. The mechanism of their action is assumed to be shown via ligand-receptor interaction with certain cell membrane receptors and via blockage of hantavirus glycoproteins (G1 and G2), resulting in adsorption inhibition and preventing viral penetration into the macrophages.

Inhibition of thrombin and factor Xa by Fucus evanescens fucoidan and its modified analogs.

Specimens of fucoidan extracted from Fucus evanescens were purified from protein and polyphenols, deacetylated and depolymerized by fucoidanase for evaluation of their biological activity. Deacetylation did not modify the capacity of fucoidan to inhibit thrombin and factor Xa, while purification from protein and polyphenols reduced this capacity. Depolymerization of fucoidan increased its capacity to inhibit thrombin mainly through heparin cofactor II. All the studied specimens formed complexes with protamine sulfate.

[Value of enzyme-linked immunosorbent assay for detection of p24 antigen of human immunodeficiency virus in confirmation of HIV-infection].

AIM: To evaluate diagnostic value of p24 antigen detection for algorithm of confirmatory diagnostics of HIV-infection. MATERIALS AND METHODS: Concurrently with Western blot assay (WB, "New Lav Blot1", Bio-Rad), tests for detection of p24 antigen of HIV (Genetic Systems HIV-1 Ag EIA", "VectoHIV-1 p24-antigen confirming test", and "DS-EIA-HIV-AG-SCREEN") were used for confirmation of first-positive result of immuno-enzyme assay. RESULTS: p24 HIV antigen was detected in serum samples in 8.4% of patients with equivocal result of WB and in 4.2% of patients with negative and positive results of WB. Presence of p24 was correlated with high viral load, and, in patients with confirmed diagnosis, with low CD4 cells count (<500 cells/ml). p24 was detected in more than 30% of persons with confirmed seroconversion after primary testing. CONCLUSION: In groups of persons with negative and equivocal results of WB assay, detection of HIV p24 antigen points to the presence of infection and could be the reason for the final diagnosis. Detection of p24 in patients with positive result of WB assay allows to consider them as probable candidates for highly active antiretroviral therapy.

[The action of Laminaria japonica extracts on 1,3-beta-D-glucanase, a digestive enzyme of the Strongylocentrotus intermedius sea urchin].

Nutritional attractiveness of the brown alga Laminaria japonica for the sea urchin Strongylocentrotus intermedius was studied. The composition of L. japonica was analyzed after one and two years of its life under natural conditions, in its seedlings, and in the alga partially degraded by natural factors. Substances extracted with various solvents were tested for the presence of inhibitors and activators of 1,3-beta-D-glucanase, a digestive enzyme of the sea urchin. Ethanolic extract of freshly harvested L. japonica was found to suppress the enzyme activity. Substances present in ethanolic extracts of the alga after one or two years of its life cycle and in the alga, partly degraded by natural factors, activated the sea urchin enzyme. This fact is in agreement with earlier natural observations concerning the nutritional attractiveness of such L. japonica samples for Strongylocentrotus intermedius.

[Screening of biopolymers from sea hydrocoles affecting the adsorption of Hantaan virus].

The biopolymers extracted from sea hydrocoles were screened and a group of sulfated polysaccharides--fucoidans having a pronounced inhibitory effect on the adsorption of Hantaan virus in the cultured Vero E6 cells was identified. The mechanism of action of sulfated polysaccharides was realized through competitive carbohydrate-specific binding to cell membrane receptors and through ligand-receptor interaction and blocking of Hantaan virus glycoproteins (G1 and G2).

[Polysaccharide and lipid composition of the brown seaweed Laminaria gurjanovae].

Polysaccharide and lipid composition of the Pacific brown seaweed Laminaria gurjanovae is determined. Alginic acid is shown to be the main polysaccharide of its biomass (about 28%); it consists of mannuronic and guluronic acid residues at a ratio of 3 : 1. The yield of water-soluble polymannuronic acid is low and does not exceed 1.1% of dry biomass. High laminaran content (about 22%) is found, whereas the yield of fucoidan is no more than 3.6%. Laminaran consists of two fractions, soluble and insoluble in cold water, their ratio is 2.5 : 1. Practically, insoluble laminaran is a linear 1,3-beta-D-glucan, and the soluble fraction was shown to be 1,3;1,6- 3-D-glucan. The oligosaccharide products of desulfation or partial acidic hydrolysis of fucoidan were studied by MALDI TOF MS; they were found to be fuco- and galactooligosaccharides. The fucoidan is suggested to be a highly sulfated partially acetylated galactofucan (Fuc/Gal is -1 : 1). The main lipid components of the dried L. gurjanovae are neutral lipids and glyceroglycolipids, whereas phospholipids are found in minor amounts. The main fatty acid components of lipids are 14 : 0, 16 : 0, 16 : 1 n-7, 18 : 1 n-7, and 18 : 2 n-6 acids.

Pharmacodynamic parameters of anticoagulants based on sulfated polysaccharides from marine algae.

Fucoidans isolated from Fucus evanescens and Laminaria cichorioides kelp can inhibit thrombin and factor Xa of the blood coagulation system. In rats, intravenous injection of fucoidans dose-dependently increased anticoagulant activity of the plasma. Fucoidans can form complexes with protamine sulfate. The observed quantitative differences in the action of fucoidans can result from different sulfation degree and the presence of various types of glycoside bonds in polysaccharide molecules.

[Polysaccharides of diatoms occurring in Lake Baikal].

Polysaccharide composition of neutral, acid- and alkali-soluble fractions of the diatoms Stephanodiscus meyerii Genkal et Popovsk and Aulacoseira baicalensis (K. Meyer) Simonsen of Lake Baikal has been studied. Neutral polysaccharides were represented by chrysolaminarans (1-->3;1-->6-beta-D-glucans). The chrysolaminaran from S. meyerii consists of the high- and low-molecular-weight fractions (40 and 2-5 kDa, respectively) and contains a large number of beta-1-->6-bound glucose residues. The chrysolaminaran from A. baicalensis is a low-molecular-weight 1-->3:1-->6-beta-D-glucan containing a small number of beta-1-->6 bonds, with mannitol being attached to the reducing unit of its chain. Acid- and alkali-soluble polysaccharide fractions are practically absent in S. meyerii. The alkali-soluble fraction from A. baicalensis is a low-molecular-weight (2-kDa) glycoprotein, the carbohydrate moiety of which is represented by a heteropolysaccharide.

[Immunostimulating and anticoagulating activity of fucoidan from brown algae Fucus evanescens of Okhotskoe sea]. / Immunostimuliruiushchaia i antikoaguliantnaia aktivnost' fukoidana iz buroi vodorosli Okhotskogo moria Fucus evanescens.

Fucoidan--nontoxic sulfated polysaccharide was isolated from brown algae Fucus evanescens in Okhotskoe Sea. Chemical analysis of the compound was performed, it was shown that fucoidan is freely soluble in water and acid solutions. Immunotropic and anticoagulating properties of the compound were evaluated in comparison with heparin. It was demonstrated that fucoidan in wide range of doses stimulated phagocytic and bactericidic activity at leucocytes of mice peritoneal exudate. Heparin on the contrary demonstrated depressive effect on these functions at high dose. It was shown that fucoidan has dose-dependent anticoagulating activity in vitro and in vivo comparable with heparin activity. The results of investigation demonstrated possibility of fucoidan application as immunomodulating and anticoagulating agent of plant origin.

Anticoagulant activity of fucoidan from brown algae Fucus evanescens of the Okhotsk Sea.

In vitro and in vivo experiments showed that anticoagulant activity of sulfated polysaccharide from Fucus evanescens (brown algae of the Okhotsk Sea) was similar to that of heparin. Anticoagulant properties of fucoidan are determined by thrombin inhibition mediated via plasma antithrombin III.

Effect of ionic carbohydrate-containing biopolymers on complement activation.

We studied in vitro effects of charged polysaccharides on the classic and alternative pathways of complement activation. The complement system was affected by substances having different charges. Our findings suggest that the conformation of polysaccharide molecules, but not their charge, plays a primary role in the interaction with C1 and C3 complement components followed by initiation of cascade enzymatic reactions.

Inhibition of complement activation by water-soluble polysaccharides of some far-eastern brown seaweeds.

Fucoidans and laminarans from Laminaria cichorioides, Laminaria japonica, Fucus evanescens, laminaran from Laminaria gurjanovae, other beta-D-glucans (translam, pustulan and zymosan) and lambda-carrageenan from Chondrus armatus were used to study the effect of water-soluble polysaccharides from seaweeds on the alternative pathway of complement (APC). beta-D-Glucans and fucoidans under study differed appreciably from each other by structural characteristics, and also by degree of purification. beta-D-glucans, on ability to bind complement, ranked in a line according to a degree of their purification. Highly purified beta-D-glucans under study did not reveal an ability to bind complement. The fucoidans were divided conventionally into three groups according to their action on APC. Highly sulfated alpha-L-fucan from L. cichorioides with the greatest activity toward APC and caused 50% inhibition of reaction of activation (RA) of APC in a concentration of 0.5-0.7 mg/ml. Opposite 50% of inhibition of lysis of erythrocytes by sulfated heterogeneous fucoidan from L. japonica was achieved with 20 mg/ml. All other fucoidans and lambda-carrageenan have activity at 6-10 mg/ml concentration. Decreasing the sulfate content from 36% up to 9% in sample fucoidans under study was not reflected practically in the 50% inhibition concentration. Apparently, the degree of sulfating of fucoidans did not influence their action on APC. But the positive influence of fucose in structure of polysaccharide was obvious.