The study of antioxidant activities of fucoidan from Laminaria japonica

Two large molecular weight fucoidan fractions F-A and F-B were obtained by water extraction and anion-exchange chromatography and then L-A and L-B with low molecular weight and uronic acid content were prepared by radical process degradation of F-A and F-B. The antioxidant activities of the fucoidan fractions on two oxygen free radicals and Cu2+-induced low-density lipoprotein (LDL) oxidation were studied. The results show that all the fucoidan fractions had excellent scavenging capacities on superoxide radical and hypochlorous acid, except the highly sulfated fraction L-B. In LDL oxidation system, low molecular weight fractions L-A and L-B exhibited great inhibitory effects on LDL oxidation induced by Cu2+, however F-A and F-B had little inhibitory effects in this system due to their large molecular weights. The present results suggest that fucoidan from Laminaria japonica is an excellent natural antioxidant and has great potential for preventing the free radical-mediated diseases.

Study on the enzymatic activity of Caspase-3 in response to alginic acid decomposing bacteria in Laminaria japonica Aresch.（Phaeophyta）

Caspase-3 is the major factor in apoptosis triggered by various stimuli, and plays a critical role during the apoptosis process. By using CaspGLOWTM fluorescein active caspase-3 staining method, caspase-3 enzymatic activities were detected in response to alginic acid bacteria in Laminaria japonica sporophytic tissues. Results showed that caspase-3 enzymatic activities were observed at 5 min after the infection. Caspase-3 enzymatic activity increased with the infection time, and had a tendency of moving from the infection site to outside. By applying caspase-specific peptide inhibitor Z-VAD-FMK, caspase-3 activation could be effectively abolished in the infected tissues. Our results indicate that programmed cell death (PCD) may be involved in the infected Laminaria japonica sporophytic tissues, and provide the evidence that defense mechanisms in algae may have similar caspase cascade events in animals.

Cryopreservation of Gametophytes of Laminaria japonica (Phaeophyta) Using Encapsulation-Dehydration with Two-Step Cooling Method

Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m^-2 s^-1 for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molLl sucrose prepared with seawater for 6 h, desiccated in an incubator set at 10℃ and 70% relative humidity for 4 h, pre-frozen at either -40℃ or -60℃ for 30 min, and stored in liquid nitrogen for 〉24 h. As high as 43% of survival rate was observed when gametophytes were thawed by placing the beads in 40℃ seawater and re-hydrated in 0.05 molL^-1 citrate sodium prepared using 30‰ NaCl 7 d later. More cells of male gametophytes survived the whole procedure in comparison with female gametophytes. The cells of gametophytes surviving the preservation were able to grow asexually and produce morphologically normal sporophytes.

Assessing the Germplasm of Laminaria (Phaeophyceae) with Random Amplified PoLymorphic DNA (RAPD) Method

Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for final amplification test. Among the total of 205 bands amplified, 181 (88.3%) were polymorphic. The genetic distance among different strains ranged from 0.072 to 0.391. The dendrogram constructed by unweighted pair-group method with arithmetic (UPGMA) method showed that the female and male gametophytes of the same cell lines could be grouped in pairs respectively. It indicated that RAPD analysis could be used not only to distinguish different strains ofLaminaria, but also to distinguish male and female gametophyte within the same cell lines. There is ambiguous systematic relationship if judged merely by the present data. It seems that the use of RAPD marker is limited to elucidation of the phylogenetic relationship among the species ofLaminaria.

Immunosuppressive Activities of Fucoidan from Laminaria japonica

Effects of fucoidan from Laminaria japonica on 2,4 dinitrochlorobenzene induced delayed type hypersensitivity (DTH) reaction and the serum levels of IgG, IgM, complement C3 and C4 were investigated in the present study. Results showed that oral administration of fucoidan at dose of 150 and 300 mg/(kg·d) for 9 days before the hapten challenge significantly inhibited 2,4 dinitrochlorobenzene induced delayed type hypersensitivity reaction; and also inhibited the humoral immunity. Serum C3 and C4 levels were markedly reduced by fucoidan at dose of 150 and 300 mg/kg; and serum IgG and IgM levels were reduced by fucoidan at dose of 300 mg/kg. The inhibitory effects of fucoidan on delayed type hypersensitivity suggested that it may be potential medication for chronic inflammatory diseases in the future.

The extraction of pigments from fresh Laminaria japonica

The pigments in Laminaria japonica was extracted with six organic solvents and analyzed in spectroscopy analysis. The extractions conditions were screened by an orthogonal test and the quantity of extracted pigments was determined spectroscopically. The results show that: (1) among the six organic solvents, acetone was the most effective one for the extraction; (2) the optimum extraction conditions were as follows: the ratio of S/M (solvent volume/material weight) was 30 ml/g; fresh seaweed was extracted 2 times in 2h; (3) the average total content of pigments was 1.85 mg/g (calculated with dry L. japonica).

Heavy Metal Biosorption Sites Studies of Laminaria japonica

The role played by the functional groups of Laminaria japonica treated by methanol and formaldehyde in biosorption of the heavy metals was investigated. Infrared spectroscopy analysis and biosorption experiments show that both carboxyl and amino groups join in the sorption; chemical treatment decreased the biosorption efficiency of the heavy metals.