Figure 1: Chromatographic traces of fucoidan from Laminaria hyperborea were obtained by charged aerosol detection monitoring 20 ul samples solubilized in deionized water at a concentration of 120 mg/ml. Sample trace is shown overlaid on baseline trace from a blank vehicle sample

Figure 2: FT-IR traces of a film formed from oven-dried fucoidan from Laminaria hyperborea. Sample was analyzed using an ATR accessory focusing on the wavelength range from 2000 to 750 cm-1

Figure 3: Cytokine expression in cultured Human peripheral blood mononuclear cells (PBMC). Cryo-preserved cells were seeded to a density of 10e4 cells/140 ul in 96 well plates and held at 37 oC for 1 h prior to treatment with test articles. Cells were then held for 24 h before centrifugation and collection of the supernatants for determination of cytokine expression. The bars represent the mean of two replicates. Dose response curves for both the fucoidan and beta-glucan were parabolic, i.e. the highest concentrations reduced cytokine expression. Therefore data is shown for the dose for each treatment that elicited the maximum expression levels. a) The endotoxin Lipopolysaccharide (LPS) at 50 pg/ml; b) Fucoidan from Laminaria hyperborea, at 10 mg/ml c) beta-glucans from yeast at 0.3 mg/ml. All values represent significant difference from control levels

Figure 4: Uptake of 2-hydroxyethyl methylacrylate particles (5x108/ml) by neutrophils isolated from mice orally dosed with experimental immunomodulators for two weeks, relative to controls. The "Combination" treatment was formulated by combining equal weights of fucoidan from L. hyperborea with yeast betaglucan; the blended test article was then administered at the doses indicated. Each point is the mean of three replicates; error bars indicate the standard error of the mean. The Least Significant Difference (LSD) is calculated from the within treatment error from the Two-Way ANOVA for p value = 0.05. a) Fucoidan compared to mushroom derived beta-glucans; b) Fucoidan compared to yeast derived beta-glucans and a combination treatment of fucoidan blended with yeast beta-glucans.*Significant difference from control at P = 0.05 level

Figure 5: IL-2 production in purified spleen cells from mice orally dosed with experimental immunomodulators for two weeks. Each point is the mean of three replicates; error bars indicate the standard error of the mean. The "Combination" treatment was formulated by combining equal weights of fucoidan from L. hyperborea with yeast beta-glucan; the blended test article was then administered at the doses indicated. The Least Significant Difference (LSD) is calculated from the within treatment error from the Two-Way ANOVA for p value = 0.05. a) Fucoidan compared to mushroom derived beta-glucans; b) Fucoidan compared to yeast derived beta-glucans and a combination treatment of fucoidan blended with yeast beta-glucans. *Significant difference from control at P = 0.05 level

Figure 6: Relative titers of antibodies raised to ovalbumin in treated mice. Mice were administered yeast beta-glucans or fucoidan from L. hyperborea by oral dosing for ten days and then inoculated with ovalbumin as antigen, followed by a booster inoculation two weeks after the initial injection. Serum was collected one week later and the titers of specific antibodies to ovalbumin were determined by ELISA. Each point is the mean of three replicates; error bars indicate the standard error of the mean. The Least Significant Difference (LSD) is calculated from the within treatment error from the Two-Way ANOVA for p value = 0.05. *Significant difference from control at P = 0.05 level

Sample	Mwa	Mnb	Mw/Mnc	IVd	Rhe	M-Haf	Recovery %
Prep 1 analysis 1	1.22E+06	337210	3.62	1.06	22.9	0.55	83.1
Prep 1 analysis 2	1.21E+06	345062	3.50	1.01	22.67	0.55	83.9
Prep 2 analysis 1	1.22E+06	364724	3.36	1.07	22.83	0.65	78.4
Prep 2 analysis 2	1.22E+06	368059	3.30	1.06	22.98	0.63	78.7
Mwa Weight average molecular weight Mnb Number average molecular weight Mw/Mnc Polydispersity index IVd Intrinsic viscosity Rhe Hydrodynamic radius M-Haf The Mark-Houwink Slope Table 1: Molecular weight determinations of the fucoidan isolate from L. hyperborea were undertaken by Gel Permeation/Size Exclusion Chromatography. Two different experimental samples were prepared from the same fucoidan manufacturing lot (Prep 1 and Prep 2) each sample was then subjected to two consecutive analyses

Experiment 1
Fucoidan	14	-13	3
Mushroom beta-glucan	9	3	1
Experiment 2
Fucoidan	11	6	10
Yeast beta-glucan	24	-5	16
Combination (fucoidan+Yeast beta- glucan)	30	-1	5
Table 2: Relative increases in the fluorescence signal of FITC tagged antibodies to Cluster of Differentiation (CD) receptors on mouse spleen cells, isolated from mice receiving 5 mg/kg test article orally, as detected under flow cytometry. Data (n = 3) is expressed as the percent increase or decrease in signal relative to untreated controls