Enzymatic extract from Ecklonia cava induces the activation of lymphocytes by IL-2 production through the classical NF-κB pathway.

Activated nuclear factor-kappa B (NF-κB), a well-known transcription factor, leads to the development, differentiation, and proliferation of T and B lymphocytes and the secretion of cytokines by the classical pathway. We have examined here whether an enzymatic extract (ECK) from the brown seaweed, Alariaceae Laminariales Ecklonia cava may contribute to activating lymphocytes through the NF-κB pathway for participation in immune responses. In our study, ECK dose-dependently enhanced the proliferation of lymphocytes. ECK significantly increased the phosphorylation of inhibitors of κB at 0.25 and 0.5 h of exposure, followed by its gradual decrease. In addition, NF-κB p65 was gradually activated, and its binding to nuclear deoxyribonucleic acid was observed from 0.25 h after stimulation (up to 0.5 h). Further experiments showed that the application of N-p-tosyl-L: -phenylalanine chloromethyl ketone, an NF-κB inhibitor, significantly blocked ECK-induced lymphocyte's proliferation and the interleukin (IL)-2 productions. Accordingly, our results suggest that ECK increases the production of IL-2 through the activation of NF-κB then induces the proliferation of lymphocytes with the coordinated stimulation of IL-2.

Detection of nitric oxide (NO) in marine phytoplankters.

In raphydophyte cell suspensions such as those of Chattonella marina, C. ovata and Heterosigma akashiwo, a gradual increase in NO-specific fluorescence intensity was observed, and the increase in the fluorescence intensity of each of these phytoplankters was completely inhibited in the presence of carboxy-PTIO, a specific NO scavenger. However, no such significant changes were observed in the case of other phytoplankter species.

Anticoagulant activity of marine green and brown algae collected from Jeju Island in Korea.

Twenty-two algal species were evaluated for their potential anticoagulant activities. Hot water extracts from selected species, Codium fragile and Sargassum horneri showed high activated partial thromboplastin time (APTT). Ultraflo extract of C. fragile and S. horneri exhibited the most potent anticoagulant activity. Furthermore, in both algal species, active compounds were mainly concentrated in >30kDa faction. The crude polysaccharide fraction (>30kDa; CpoF) of C. fragile composed of approximately 80% carbohydrate and approximately 19% of protein; the crude polysaccharide fraction (>30kDa; CpoF) of S. horneri was composed of 97% of carbohydrate and approximately 2% of protein. Therefore, most probably the active compound, or compounds of the algal species were related to high molecular weight polysaccharide, or a complex form with carbohydrate and protein (proteoglycan).

Partial characterization and immunostimulatory effect of a novel polysaccharide-protein complex extracted from Phellinus linteus.

Many polysaccharides isolated from mushroom are considered to be biological response modifiers and have been shown to enhance various immune responses in vivo and in vitro. We demonstrate that a novel polysaccharide-protein complex (PPC) extracted from Phellinus linteus was a potent immunomodulator. PPC had a molecular weight of approximately 73 kDa. It was composed of five different monosaccharides, predominantly D-glucose and D-mannose, in the molar ratio of 3:2, the main amino acid being aspartic acid. PPC had a unique mode of immunostimulation with regard to its cell-type specificity. PPC was found to markedly increase the proliferation of B cells, but not T cells. Although PPC and lipopolysaccharide (LPS) had a similar mode of action in B cells, they were differentiated by the fact that PPC-induced cellular activation was not inhibited by polymyxin B (PB), a specific inhibitor of LPS. PPC increased the cytokine production and nitric oxide (NO) from macrophages. PPC also enhanced the lytic death of NO-sensitive tumor cells, B16 melanoma, through the production of NO. In addition, PPC up-regulated the natural killer (NK) cell-mediated killing of tumor cells, YAC-1 lymphoma in vitro. These results suggest that PPC stimulated the tumoricidal activities of macrophages and NK cells, and induced the proliferation of B cells in vitro. This process may be the mechanism by which PPC produced its therapeutic effects.

Antioxidant activities of enzymatic extracts from brown seaweeds.

Potential antioxidative activities of enzymatic extracts from seven species of brown seaweeds were evaluated using four different reactive oxygen species (ROS) scavenging assays containing DPPH (1,1-diphenyl-2-pricrylhydrazyl) free radical, superoxide anion, hydroxyl radical and hydrogen peroxide scavenging assay. The brown seaweeds were enzymatically hydrolyzed to prepare water-soluble extracts by using five carbohydrate degrading enzymes (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme and Alcalase) of commercial and inexpensive enzymes obtained from Novozyme Co. (Novozyme Nordisk, Bagsvaerd, Denmark). The enzymatic extracts exhibited more prominent effects in hydrogen peroxide scavenging activity (approximately 90%) compared to the other scavenging activities and the activity of enzymatic extracts was even higher than that of the commercial antioxidants. In particular, Ultraflo and Alcalase extracts of S. horneri were dose-dependent and thermally stable. Moreover the two enzymatic extracts strongly inhibited DNA damage (approximately 50%). Those extracts showed significantly (p<0.05) remarkable scavenging effects in DPPH free radical scavenging assay and the activity indicated a marked correlation with phenolic contents. From the results, enzymatic extracts of the brown seaweeds might be valuable antioxidative sources.