Irina M. Yermak

Comparative study of carrageenans from reproductive and sterile forms of Tichocarpus crinitus (Gmel.) Rupr (Rhodophyta, Tichocarpaceae)

A comparative study of the structure and properties of the sulfated polysaccharides (carrageenans) isolated from the vegetative and reproductive forms of the red alga Tichocarpus crinitus was performed. The polysaccharides were separated into the gelling (KCl-insoluble) and non-gelling (KCl-soluble) fractions by precipitation with 4% KCl. The total content of polysaccharides extracted from the reproductive form of the alga was 1.8-fold more than that extracted from the vegetative form, and in the first case, the gelling polysaccharides mostly accumulated. The gelling polysaccharides from the vegetative form have the highest molecular weight (354 kD). According to the results of FT-IR and 13C-NMR spectroscopy, the gelling polysaccharide fractions from both forms are κ/β carrageenans. The differences concern the content of the κ- and β-disaccharide units and the presence of a small content of the sulfated disaccharide segments (precursors of the κ-carrageenans) in the polysaccharide from the reproductive form of the alga. The non-gelling polysaccharide fractions from both forms of the plant are mixtures of sulfated galactans with a low content of 3,6-anhydrogalactose.

Carrageenans from cystocarpic and sterile plants of Chondrus pinnulatus (Gigartinaceae, Rhodophyta) collected from the Russian Pacific coast

The chemical structure, gel properties and biological activity of the carrageenans isolated from cystocarpic and sterile plants of Chondrus pinnulatus were investigated. The total carrageenan content of the sterile plant was observed to be twice that of the cystocarpic plants. According to data obtained by 13C-NMR and FT IR, the gelling polysaccharides from cystocarpic and sterile plants of C. pinulatus have similar structures and were identified as κ/ι-carrageenans. The difference between these polysaccharides was in the ratio of the κ- and ι-segments, with a predominant content of κ-segments in cystocarpic plants (80%). Moreover, KCl-insoluble fractions possibly contain hetero-disperse μ/ν precursor: amounts of this in the polysaccharide from sterile plants were more than that extracted from the cystocarpic plants. The KCl-soluble fractions (non gelling) were λ-carrageenans with another carrageenan type that had a low amount of 3,6-anhydrogalactose. Carrageenans from cystocarpic stages showed good gelling properties, whereas those from sterile plants formed a very weak gel. Structural differences and molecular weight of carrageenans obviously determine the biological activity of the polysaccharides. Non gelling-carrageenans from both types of ιt C. pinnulatus plants showed high macrophage-phosphatase activity and κ/ι-carrageenan from cystocarpic plant possessed a potent anti-coagulant activity, which was extremely strong in a low concentration of 100 μg ml−1.

Effect of κ/ß-Carrageenan from red alga Tichocarpus crinitus (Tichocarpaceae) on infection of detached tobacco leaves with tobacco mosaic virus

It is shown that κ/ß-carrageenan obtained from red marine alga Tichocarpus crinitus possesses antiviral activity against Tobacco mosaic virus (TMV). Two days after inoculation of Nicotiana tabacum L. cv. ‘Samsun’ detached leaves with a mixture of TMV (1 μ/ml) and carrageenan (1 mg/ml), the infectivity and virus content in the leaves as well as the intracellular accumulation of virus particles were significantly inhibited in comparison with leaves inoculated with TMV only. The carrageenan applied 24 h prior to inoculation with TMV also exhibited the antiviral effects but to a lower degree than when the virus and polysaccharide were inoculated together. As the infection developed, the inhibitory effects of carrageenan decreased. Treatment of the leaves with carrageenan affected the formation of TMV-specific intracellular inclusions (granular and tubule) known to consist of the virus-encoded protein components of the viral replicase. A characteristic of the infection development in the presence of carrageenan, 4 days after infection of the leaves with TMV, was the predominant formation in cells of granular inclusions famous to appear at early stages of TMV multiplication. At the same time, cells of the leaves inoculated with TMV alone contained for the most part tubule inclusions considered to arise from granular ones on the advanced infection stages. Taken together, our observations indicate that the κ/ß-carrageenan inhibits TMV infection in detached tobacco leaves at early stages.ZusammenfassungEs wurde gezeigt, dass κ/ß-Karagheen, das aus den roten Meeresalgen Tichocarpus crinitus gewonnen wurde, hohe Antivirusaktivität gegen das Tabakmosaikvirus (TMV) besitzt. Zwei Tage nach der Inokulation der Blätter von Nicotiana tabacum L. cv. ‘Samsun’ mit der Mischung TMV (1 μ/ml) und κ/ß-Karagheen (1 mg/ml) werden die Infektionsstufe, der Gehalt an Virus in den Blättern, und auch die Akkumulation von Virusbestandteilen in den Zellen signifikant gehemmt. κ/ß-Karagheen, das 24 h vor der Inokulation mit TMV angewendet wurde, auch Antiviruseffekte, aber weniger als wenn Virus und Polysaccharid gemeinsam inokuliert wurden. Im Laufe der Entwicklung der Infektion wurden die Hemmeffekte von κ/ß-Karagheen immer schwächer. Die Behandlung der Blätter mit κ/ß-Karagheen beeinflusste die Entstehung von TMV-spezifischen intrazellulären Komponenten (granulär und tubulär), die bekanntlich aus den virus-codierten Eiweißkomponenten der Virenreplikase bestehen. Charakteristisches Merkmal der Entwicklung der Infektion in Anwesenheit von κ/ß-Karagheen (4 Tage nach der Inokulation der Blätter mit TMV) war die vorwiegende Bildung der Granular- komponenten in den Zellen, die in den Frühstadien der TMV-Replikation entstehen. Die Zellen der Blätter, die nur mit TMV inokuliert worden waren, enthielten hauptsächlich die tubuläre Komponente, die, wie man vermutet, sich während der fortschreitenden Etappen der Infektion aus den Granular- komponenten bilden. Die Ergebnisse zeigen, dass κ/ß-Karagheen die TMV-Infektion in abgeschnittenen Tabakblättern hemmt.

Structure and properties of carrageenan-like polysaccharide from the red alga Tichocarpus crinitus (Gmel.) Rupr. (Rhodophyta, Tichocarpaceae)

Sulfated polysaccharides occurring in the red algae Tichocarpus crinitus cell wall were fractionated and purified. NMR and FT-IR spectroscopy analyses revealed that the non-gelling fraction contained a sulfated galactans having a new carrageenan-like structure. It is built with alternatively linked 1,3-linked β-D-galactopyranosyl-2,4-disulphates and 1,4-linked 3,6-anhydro-α-D-galactopyranosyl residues. Minor amounts of its biosynthetic precursor were detected in a water-extracted specimen. Brief analysis of rheological and biological properties of the non-gelling fraction was carried out. The carrageenan-like polysaccharide from T. crinitus displayed the properties of “random coil” polymer at high temperature, and possesses high anticoagulant activity at low concentration.

Marine Compounds with Therapeutic Potential in Gram-Negative Sepsis

This paper concerns the potential use of compounds, including lipid A, chitosan, and carrageenan, from marine sources as agents for treating endotoxemic complications from Gram-negative infections, such as sepsis and endotoxic shock. Lipid A, which can be isolated from various species of marine bacteria, is a potential antagonist of bacterial endotoxins (lipopolysaccharide (LPSs)). Chitosan is a widespread marine polysaccharide that is derived from chitin, the major component of crustacean shells. The potential of chitosan as an LPS-binding and endotoxin-neutralizing agent is also examined in this paper, including a discussion on the generation of hydrophobic chitosan derivatives to increase the binding affinity of chitosan to LPS. In addition, the ability of carrageenan, which is the polysaccharide of red alga, to decrease the toxicity of LPS is discussed. We also review data obtained using animal models that demonstrate the potency of carrageenan and chitosan as antiendotoxin agents.

Atomic force microscopy imaging of carrageenans from red algae of Gigartinaceae and Tichocarpaceae families

In the present article, the atomic force microscopy was applied to investigate macromolecular structures of various carrageenan types including hybrid polysaccharides (κ-, κ/β-, κ/ι-, λ-, and X-carrageenans) depending on polysaccharide concentration. The structures dependence on a polysaccharide concentration also was focused. κ-Carrageenan forms both single and two stranded structures at a low concentration. At high concentrations κ-, κ/β-, and κ/ι-carrageenans form fibrous network-like structures by a side-by-side association type at the same time for κ/ι-carrageenan end-to-end association type also was found. Comparably to κ-carrageenan, κ/β-carrageenan network was more open with coarser fibers while κ/ι-carrageenan structure is characterized with a more flexible network. Honeycombed structures due to end-to-end and side-by-side association types were observed for X-carrageenan, while λ-carrageenan formed honeycombed structures only at high concentrations. In order to investigate topographical parameters of the carrageenans macromolecular structure a new method of the autocorrelation function analysis was used for the first time.

Influence of κ/β-carrageenan from red alga Tichocarpus crinitus on development of local infection induced by tobacco mosaic virus in Xanthi-nc tobacco leaves

The influence of κ/β-carrageenan from red marine alga Tichocarpus crinitus on the development of tobacco mosaic virus (TMV) infection in Xanthi-nc tobacco leaves was studied. It was shown that the number of necrotic lesions on the leaves inoculated with the mixture of TMV (2 μg/ml) and carrageenan (1 mg/ml) was reduced by 87%, compared to the leaves inoculated with the virus only. The suppression of virus infection was also observed when leaves were treated with carrageenan 24 h before or 24 h after leaf inoculation with TMV; however, in these cases, suppression was less evident than after inoculation with the virus-polysaccharide mixture. It is supposed that the antiviral activity of carrageenan applied together with TMV may be explained by its action not only on the plant but also on the virus itself. The inhibitory effect of carrageenan pretreatment can be explained by its favorable effect on tissue resistance to infection. The suppression of this resistance by actinomycin D indicates that carrageenan functions via its action on the cell genome.

Soluble chitosan–carrageenan polyelectrolyte complexes and their gastroprotective activity

The soluble polyelectrolyte complexes (PEC) κ-carrageenan (κ-CG):chitosan was obtained. Binding constant value (2.11 × 10(7)mol(-1)) showed high affinity of κ-CG to chitosan. The complex formation of κ-CG:chitosan 1:10 and 10:1 w/w was shown by centrifugation in a Percoll gradient. Using atomic force microscopy we showed that the supramolecular structure of the complexes is different from each other and from the macromolecular structure of the initial polysaccharides. The gastroprotective and anti-ulcerogenic effect of κ-CG, chitosan and their complexes was investigated on the model of stomach ulcers induced by indometacin in rats. PEC κ-CG:chitosan have gastroprotective properties which depend on their composition. Complex κ-CG:chitosan 1:10 w/w possesses higher gastroprotective activity than the complex 10:1 w/w. These results suggest that the gastroprotective effect of complexes can be associated with their protective layer on the surface of the mucous membrane of a stomach, which avoids a direct contact with the ulcerogenic agent.

In Vitro and Ex Vivo Studies of Antioxidant Activity of Carrageenans, Sulfated Polysaccharides from Red Algae

Antioxidant properties of structurally different sulfated polysaccharides (carrageenans) were studied in vitro and ex vivo. Ferric reducing antioxidant activity of carrageenans and their inhibitory effects on hydroxyl radicals and superoxide anion radicals were demonstrated in vitro. Activity of carrageenans depends on the polysaccharide structure. Carrageenans stimulate catalytic activity of SOD from donor erythrocyte.

Influence of red algal polysaccharides on biological activities and supramolecular structure of bacterial lipopolysaccharide

The present work reveals the effect of kappa-, lambda- and kappa/beta-carrageenans on the immune modulation and supramolecular structure of lipopolysaccharide (LPS). The kappa/beta carrageenan was able to increase the synthesis of anti-inflammatory interleukin-10 (IL-10) in vitro, and at low concentrations, their activity in the mixture with LPS was higher than that of LPS alone. Kappa-carrageenan significantly inhibited LPS-induced upregulation of reactive oxygen species (ROS). The activation of cells by kappa-carrageenan occurs through TLR4 receptor specific for LPS. Carrageenans reduced (kappa-) or completely inhibited (lambda-) collagen-induced platelet aggregation and decreased their aggregation activity caused by cooperative effect of LPS and collagen. The transformation of ultrastructure of LPS by action of kappa- and kappa/beta-carrageenans is observed. Moreover, carrageenans changed both the sizes and ζ- potentials of the LPS.