-Regarding Content of Alginic Acid of Brown Seaweed by Part and Its Physiological Function

-Regarding Contents of Carbohydrate of SEAHERB FUCOIDAN and PURE SKIN and Its Physiological Function

I Introduction

Observing the content of formation of marine plants, generally, the content of carbohydrate is high, and especially the blossom of algae, based on dried items, has a distinctive feature to contain up to about 67% of carbohydrate.(National Fisheries Promotion Board, 1989;Oishi, 1944; Cho, et al.,1955)

And, these multi-sugars of carbohydrate contained in the blossom of algae are mostly formed with Fucoidan(about 70-80%), Laminaran and Alginic acid (Nishide, 1981 ; Aisha, 1944; Kim, et al., 1955), and there is a difference in content of Fucoidan and alginic acid of the brown algae in accordance with by part and, harvest time and place.(10-30%)

It is well known that Fucoidan, Laminaran and Alginic Acid hardly absorbed by the body (7%) due to the fact it is a giant molecule.

And, it is the situation that development of new extract processing method to increase absorption rate and producing products are keenly required.

Sea Herb Co., Ltd. has developed the SEAHERB FUCOIDAN of the new extract processing form, called “low molecular weight compound technology,” with increasing absorption rate up to 57%, with the raw material of the 15 species of seaweeds and herbs

Of this nutrition, regarding SEAHERB FUCOIDAN which is natural supplement, the writer would like to write its physiological function as follows:

2. PHYSIOLOGICAL FUNCTION OF ALGINIC ACID of SEAHERB FUCOIDAN

In order to study the fundamental knowledge on alginic acid, observing its structure, it is as Fig.2.

Alginic Acid is contained only in the seaweed, which exists as the cell wall or cell distance substance and differs in accordance with the kind of seaweed, but is contained up to about 20-50%, and compared with animals, it functions an important role to form physique.

a) Function of Heavy Metal Discharging Outside the Body

It is already well known that the seaweed plays a role to concentrate various metal ions etc., in the sea water by themselves. The fact that these seaweed absorb and discharge outside the body especially heavy metal, etc. from human body, is widely examined and recognized in the academic circle. This function is explained as Fig.3

Sr existing in diet or blood, and bone combined with NaAlg and Na is separated, and heavy metal combines with alginic acid, which is extracted outside the body as combined substance, which does not function. (Fumio Kasahara, et.al)

That is heavy metals, Cu, Pb, Co, and Zn are effected to be extracted outside the body by such principle.

On these days when the environmental pollution in our daily life has become very serious problem, if we take a little quantity of these substances from the air or diet, these will be accumulated in our body for a long time and we may face with difficulty before we knew it.

As explained as above, since alginic acid in most hardly absorbed by the body. Alginic acid inside of the SEAHERB FUCOIDAN can be absorbed by the body up to 57%. By using this low molecules alginic acid, Dong Oh international Co., Ltd., develop PURE SKIN which is skin care products, with moving through the skin rapidly. Main function of PURE SKIN is discharging heavy metal and toxin outside of the skin.

b) Decreasing the Value of Cholesterol

It has been well known that the seaweed is the ageless food since old times and as a result of experiment by Suzuki, et. al., that by adding 5% of alginic acid to the extremely inferior feed adding 0.25% of bile acid to the food containing 1% of cholesterol, let white mouse take it, and, obtained the effect to control increase of serum cholesterol and cholesterol in the blood plasma. Beside this, many other researches have been reported, so it is well known fact that alginic acid has an effect to prevent artery hardness.

c). Function of Intestinal Orders

There is nothing more important than smooth absorption of nutritive elements by smooth excretion among various functions of human body. This alginic acid is the sticky component, and it demonstrates concrete effect of internal orders by such nature as hydration, maintaining water, lubrication and formation. Especially, it has an excellent effect for constipation.

When alginic acid is degraded by enzyme and concentrated, a portion becomes ppt and a portion makes heavy metal and other undesirable things to suspended condition of their activities in the intestines and to discharge outside the human body. That is, this function is to discharge heavy metal and other undesirable things existing inside the stomach and intestines as scale condition outside the human body by the function of alginic acid and enzyme.

Therefore, it has a great effect for the health of the stomach and intestine, and especially for constipation. And if you take it steadily, it will improve digestion and absorption, and accordingly, all physical organs become healthy and have excellent function for prevention of geriatric diseases. Although we think to cure only a special portion of our body, it is thought that by making our stomach and intestines healthy first of all, and by helping to supply nutritive substance to other organs, the balance of whole body can be maintained.

d) The Diet Nature

Because of shorted calorie in our modern life, if we take much quantify of meats, fat and other alien substances are absorbed, and if you take this for a long time to prevent satiation, by digesting and absorbing in balance, so by removing unnecessary fat and energy source, it is well known as food of diet nature.

3. Carcinogenesis Restraint Effect by Fucoidan Component Contained SEAHERB FUCOIDAN

Especially a great quantity of fucoidan is contained in blossom of algae of SEAHERB FUCOIDAN, for which a steady study has been continued. As a result of giving cancer-causing materials to the experimental mice, which were divided into two groups, and let one group drank water containing U-Fucoidan for 52 weeks continuously, and the result was confirmed that the group drank U-Fucoidan water increased about 30% of surviving percentage and reduced about 10% of carcinogenesis percentage compared with the comparative group.

4. RESULT SUMMARY

The Nutrients of the SEAHERB FUCOIDAN is greatly used in our dietary life, and it is a very good supplement because a great quantity of Fucoidan and alginic acid is contained in the SEAHERB FUCOIDAN, which functions to absorb and discharge outside our body heavy metal and other undesirable things.

As the contents of fucoidan and alginic acid in the seaweed differs in accordance with the producing season, place and by location. It is possible to choose the season, place, and location and part having the greatest content of Fucoidan and alginic acid.

These Nutrients are enhanced by our patent extracting technology called “Low Molecular weight compound technology” which has the possibility of the function of discharging heavy metal outside of the body, the function of decreasing the value of cholesterol, function of intestinal orders, and the nature of diet.

Furthermore, Dong Oh International co., ltd adjusts extracting technology for developing PURE SKIN, which is a skin care products, for discharging toxin and heavy metals outside skin and providing vitality of skin.

The SEAHERB FUCOIDAN, which is processed to 100% natural pill type without adding any artificial preservatives, color and corn in the sealed bottle package and can be carried to take at any time, is expected to greatly contribute to the health of the people.

1. Fumio Kasahara:Regarding Physiological Function, New Food Industry vol. 22 No.12

2.Eiichi Nishide, et al:Distribution by part of water Soluble Alginic Acid and Alkali soluble Alginic Acid of hot water extract from Brown Seaweed Nippon Suisan Gakkaishi 54(9),1619-2622(1988)

3.Kazuhiro Watanabe, etal:influence to Excretion of Rat by cholesterol of Alginic Acid Natrium, Trp-p-1 and Afratoxin B, Toicold Environ Health 38(3) 258-282(1992)

4.Tokuichi Tabuchi: Algin and Its Use, New Food Industry vol.20, No.9.

New Wave Of Health From The Sea

By Dr. Zakir Ramazanov, President of National Bioscience Corporation, Warwick, NY

Sea vegetables offer untapped plethora of health benefits. Much attention has been focused of late on the Asian diet, which appears to contain a wealth of protective health-promoting compounds. An examination of the Asian diet reveals that it is rich in sea vegetables. It should come as no surprise, therefore, that the long recognized traditional health benefits of certain sea vegetables are now being confirmed by modern scientific research.

Seaweed has long been used in the Japanese and Chinese diet. In 600 B.C., Sze Teu wrote in China,
"Some algae are a delicacy fit for the most honored guests, even for the King himself." Some 21 species are used
in everyday cooking in Japan , six of them since the eighth century. Seaweed accounts for some 10% of the
Japanese diet and seaweed consumption reached an average of 3.5 kg per household in 1973, a 20% increase
in 10 years. Most important are nori (Porphyra species), kombu (Laminaria spp.) and wakame (Undaria spp.).

In the West, seaweed is largely regarded as a health food and, although there has been an upsurge of interest in the last 20 years, it is unlikely that sea vegetable consumption will ever be more than a fraction of what it is in Japan.
Among U.S. suppliers, all sea vegetables are known under a very general name "Pacific" and/or "Atlantic" kelp, which represents in most cases a "cocktail" of a dozen plant species harvested in the open ocean.

The focus of scientific inquiry has been directed at a sub-group of sea vegetables known as phaeophytes (brown seaweed). Powerful antioxidant activity was recently discovered in certain members of this class and this finding
has resulted in an enormous increase in research on sea vegetables' metabolites and their activity against free
radicals. The fruits of this research have shown that certain phaeophytes contain highly active polyphenols called
phloroglucinols (1,3,5-trihydroxybenzene) that possess free radicals neutralizing activity.

Recently Japanese scientists discovered that phenolic compounds such as phloroglucinols possess 5-lipoxigenase activity. The products of arachidonic acid metabolism are not tumor promoters themselves,
but play an integral part in the underlying mechanism of tumor promotion and 5-lipoxigenase is a key enzyme
in the arachidonic cascade. Phytoactives present in brown sea vegetables possess antibacterial activity and
phloroglucinol is responsible for this effect. In addtion phloroglucinols are moderate MAO inhibitors.
This fact could contribute to heir strong t anti-depressant activity.

Sea vegetables and cholesterol. The Japanese have believed for many years that eating seaweed prolongs life. Since many deaths are due to heart disease, which mav he linked with high plasma cholesterol levels and hypertension, physiological the effect of seaweed on these values have been investigated.
Again, sea vegetables such as Ascophyllum, (Cystoseira and Fucus have been shown to lower significantly
plasma cholesterol levels (Krotkiewski M., European Patent #90850263.6) and the active compounds have been identified: Fucosterol and the unsaturated fatty acids show hypocholesterolemic activity.

This ability to reduce plasma cholesterol levels and to increase serum lipolytic activity may explain their use in the prevention of atherosclerosis. An antihypertensive activity of substances with sodium-binding properties,
e.g. a polysaccharide, is obtained from brown seaweed fibers.

Sea vegetables and cancer. Certain sea vegetables have long been used in traditional Japanese and Chinese medicine in the treatment of cancer. Oxidative processes are involved in both the initiation of carcinogenesis and
the promotion of tumor development (Pryor, 1987). Research of recent years provides strong evidence that the sea vegetables Ascophyllum, Cystoseira and Fucus showed antitumoral activity (lC50mg/mL) against leukemia P-388 (Norte et al. 1995).

Based on epidemiological and biological data, consumption of sea vegetables are proposed as an important factor contributing to the relativelv low breast cancer rates reported in Japan . It is well known that sea vegetables are the best source of nondigestible fiber, increasing fecal bulk and decreasing bowel transit time;
it changes the posthepatic metabolism of sterols; it contains an antibiotic substance that may influence fecal ecology; it contains 1-3 beta glucan, which alters enzymatic activity of fecal flora and it stimulates the
host-mediated immune response.

It is suggested that Laminaria may play a role in preventing either the initiation of breast cancer or its promotion
by endogenous physiological factors. Among mammary adenocarcinoma tumor-bearing animals, experimental
rats had fewer individual adenocarcinomas. There was also an overall 13% reduction in the number of experimental rats with histologically confirmed denocarcinomas (76% among the control rats compared to 63% among the experimental rats).Components of Laminaria, which might account for the observed difference in mammary tumor
growth are varied and include the sulfated polysaccharide Fucans or fucoidan.

Fucans, sulfated polysaccharides extracted from brown seaweed, have been shown to have inhibitory effects on cell growth in various experimental models. These findings raise the possibility that brown seaweed may,
have clinical value in the prevention of cancer metastasis.

Sea vegetables commonly eaten in Japan inhibit breast carcinogenesis.
Anti-mutagenic activities were detected in polysaccharide and non-polysaccharide fractions from the extract of the
other edible brown alga, Undaria pinnatifida (wakame in Japanese).
These experimental results indicate that the hot water-soluble extract of Larninaria japonica or Undaria pinnatifida
contains heterogenous antimutagenic activities against typical genotoxic substances.

The role of seaweed in breast cancer treatment was indirectly implicated at the inception of the low thyroid breast cancer hypothesis. Ancient Egyptians gave seaweed to breast cancer patients (Ebers Papyrus) and suggested
that the iodine content in the seaweed was responsible for stimulating the thyroid. This hypothesis has generated many studies over the last 25 years, although none were able to establish a causal link between thyroid dysfunction
and subsequent breast cancer.

It has been observed that women with breast cancer who also have thyroid dysfunction have a poorer prognosis
for both five and ten year survival.
The most convincing data arguing against this theory is that as a result of iodized salt, endemic goiter rates have declined in the U.S. , while breast cancer mortality rates have not.
It is therefore intriguing to consider the alternative explanation -that it was the seaweed rather than the iodine
content that lent efficacy to the Egyptian treatment.
The existing data are supportive of the idea that sea vegetable consumption as well as terrestrial vegetables might
be a protective factor.

Sea vegetables and HIV Anti-HIV-active and anti-herpes polysaccharides and polyphenols have been isolated in brown seaweed Fucus vesiculosus and red alga Dumontia.
What about green sea vegetables such as Ulva rigida or sea lettuce, an edible alga containing the extraordinary
potencies of soluble and insoluble dietarv fiber and important minerals, as well as high potencies of vitamins, polysaccharides, chlorophyll and protein? Ulva rigida is a member of the group of green seaweeds Monostroma
and Enteromorpha, which are called "annori" in Japanese.

Ulva has been used in salad and soups for many centuries; however, the recent discovery that xylorhamnoglucuronan (Ulvan) and glucuronic acid are major constituents of Ulva dramatically increased interest
in this seaweed. Glucuronic acid is a strategically important component of chondroitin (viscosity mucopolysaccharides that act as the flexible connecting matrix between collagen filaments in cartilage to form
a polymeric system). To be able to svnthesize enough chondroitin, the human body needs glucuronic acid and glucosamine. The lack of one of these components dramatically decreases the chondroitin synthesis.

Russian scientists provided strong evidence that the rate of CI4 chondroitin sulfate synthesis de novo is much
higher from I 4C glucuronic acid (Ulva) than from Cl4 chondroitin (Sidorov et al. 1977; Applied Microbiology and Biotechnology). We believe that chondroitin is hydrolyzed before it becomes bio-available in the body and
that the rate of this hydrolysis not so high as to compete with free glucuronic acid (Sidorov et al. 1977).
These results are not surprising, since glucuronic acid is an essential constituent of chondroitin.
Beside glucuronic acid, Ulva rigida contains high proline and ascorbic acid required for collagen synthesis.

In the process of collagen synthesis two amino acids, proline and hydroxyproline, play a strategically important role. Collagen synthesis is initiated by hydroxylation of proline, which requires the presence of a sufficient amount of proline and ascorbic acid - a source of OH-groups needed to transform proline into hydroxyproline.
Ulva rigida provides both proline and ascorbic acid.

Bibliography

Fujimoto I., Hanai, A. and Oshima, A. Descriptive epidemiology of cancer in Japan : current cancer incidence and survival data. National Cancer Institute Monograph 53: 5-15.1979.

Kagawa, Y. Impact of Westernization on the Japanese. Changes in physique,
cancer longevity and centenians. Preventive Medicine 7: 205-217.1978.

Katsura, E. and Nakamichi, R. The iodine intake of Japanese. Journal of Japanese Society for Food and Nutrition
12: 37-39.1960.

Kimura, A. and Kuramoto. M. Influences of seaweeds on metabolism of cholesterol and anticoagulant actions of seaweed. Tokushima Journal ofExperimental Medicine 21:79-88. 1974.

Michahek, G. Seaweed Resources of the Ocean FAO Fisheries Technical Paper, No. 138: 79-98. 1976.

Moossa, A. R., Price Evans, D. A. and Brewer, A. C. Thyroid status and breast cancer,
Annals of the Roval College Surgeons England 53:178-188. 1973.

Morgan, K C., Wright, J. L. C. and Simpson, F J. Review of chemical constituents of the red alga.
Palmaria palmata (Dulse). Economic Botany 34(1): 27-8T.1990.

Pratt, R., Mautner, H., Gardner, G. M., Sha, Y and Dufrencoy, J. Report on antibiotic activity of seaweed extracts. Journal of the American Pharmaceutical Association 40 (11): 575-579.1951.

Teas J. The consumption of seaweeds as a protective factor in the etiology of breast cancer,
Medical Hypothesis 7,601-613, l98l.

Toyokawa, H. Nutritional status in Japan from the viewpoint of numerical ecology Social Science and Medicine
12(I): 517-524.1978.

Vacca, D. D. and Walsh, R. A. The antibacterial activity of an extract obtained from Ascophyllum nodosum.
Journal of the American Pharmaceutical Association 43:24-26. 1954.

Wood, C. G. Seaweed extracts a unique ocean resource.
Journal of Chemical Education 51(7): 449-452. 1974. NW

Seaweeds or marine algae have long made up a key part of the Asian diet and are also consumed in other parts S of the world, such as in Ireland and Wales. Seaweed has often been used as a food for people who are sick and has been credited with health-giving properties.1 Today, seaweed supplements for human use are usually considered to be sources of iodine or minerals but may offer other therapeutic
benefits.

Marine Algae as Food

Marine algae are classified as brown, red, or green algae.2 Examples from all of these categories are edible and are shown in Table 1. This article concentrates on brown
seaweeds, in particular the commonly eaten Japanese wakame or Undaria pinnatifida.

Japanese and Korean populations are the biggest consumers in e world of seaweed products. Most of the brown seaweed intake in the daily diet is of Undaria, commonly known as e and mekabu and of Laminaria species commonly known as kombu.1,2 Both are used dried in condiment and soup bases or eaten fresh in salads, rolls, or stews, or with rice.

It is thought that the overall content of certain traditional diets contributes to the low incidence of cancer,3 particularly breast cancer.4 It is apparent that the unique levels of seaweed intake contribute to the variance in the levels of breast cancer.-5,6 There is a nine fold lower incidence of breast cancer in the Japanese population and an even lower incidence the Korean population compared to the incidence
in the West.4,5,7

The relative longevity and health of Okinawan Japanese populations has been attributed in part to dietary algae in studies.8 these studies compared Okinawan descendants who were living in Brazil with Okinawans. The former have a higher risk of developing cardiovascular and other diseases. For a dietary intervention study, 3g of decosahexaenoic acid, 5g of seaweed wakame) powder, and 50 mg
of isoflavonoids from soybean Glycine soja) were given daily to immigrants, at high risk for eloping diseases, in Brazil for 10 weeks. This combination reduced blood pressure and cholesterol levls, suppressed the urinary markers of bone resorption, and attenuated a tendency toward diabetes.

Contents of Algae and Algal Extracts

Brown algae consist mainly of water (90 percent) in the native state. Polysaccharides
are major components and comprise alginates, cellulose, and sulfated polysaccharides such as fucoidans and laminarins. Other components include proteins, free mannitol, minerals such as iodine and arsenic (inorganic and organic), polyphenols, peptides, fatty compounds, and various pigments.1 Alginates, probably the most widely used of the algal extracts, are composed of block copolymers of mannuronic and guluronic acid sugars and have been adopted by the food industry as thickening agents and by the pharmaceutical industry as binders, gelling agents, and wound absorbents.

Possible Therapeutic Role of Sulfated Polyanions

Fucoidans are found only in brown algae. They consist of long branched chains of sugars and include a substantial amount of fucose. The type of fucoidan, its sulfation, molecular weight, and conformation of sugar residues varies with the species of sea-weed.1,9,10 Thus, the fucoidan from Fucus vesiculosis contains 90 percent fucose,ll whereas the major fucoidan in Undaria contains a roughly equal balance of fucose and galactose.12,13

When purified, the fucan fractions from Korean Undaria and Laminaria comprised respectively 12.75 and
4.76 percent of the total dry weight of the algae.12 The fucose composition (as a per- cent of total sugars) of the fucoidans isolated from the Undaria and Laminaria was 57.11 percent and 80.43 percent respectively.
It should be noted that while the bulk of the fucoidans in seaweeds are of high molecular weight, there is a small percentage of smaller fucoidan-type molecules that are sometimes complexed with proteins.11,13

Fucoidans are considered to have similarities to the (much smaller) mammalian molecule heparin sulfate.14,15 As such, they compete for heparin sulfate-type receptors such as those used for viral entry into cells.16,17 Thus' fucoidans are highly effective antiviral agents.14,15 In addition, they inhibit leukocyte movement into tissues,18 modulate metastasis,19 have heparin-like anticoagulant qualities in vitro,20 anticomplement activity in vitro and in vivo,21,22 and antilipidemic activity in vivo.13 Other biologic activities related to the heparin-like nature of fucoidans
include stimulating hematopoietic progenitor cell mobilization23,24 and inhibiting smooth-muscle proliferation.25

Most of the effects noted above were observed when using intravenous fucoidans in animal models.
However, in vivo biologic effects after Undaria ingestion indicate that there is uptake of active components,
via the gut; for example, ingestion of water extracts of Undaria by mice with carcinogen induced tumors markedly suppresses tumor development.6,26 Hiebert27 has found that oral heparin is absorbed and found at approximately
I percent in plasma in rats. There is a pronounced endothelial uptake of oral heparin and, thus, oral heparins
have marked bio- logic effects despite the low plasma levels. It is possible that fucoidans behave similarly.

Use of Algae in Traditional Medicines

Chinese and Kampo (Japanese) medicine both use dried thallus (stem and spore areas) of brown seaweeds (Laminaria, Undaria, or Ecklonia species).
These are used to "eliminate phlegm and move water" and are also recognized sources of iodine.28

They are recommended for treating cancer in Chinese and Ayuverdic medicinal
texts.28,29

In Korea , new mothers are given a diet that is rich in seaweed for the first month after birth because this diet is believed to provide many health benefits for mothers and their children.30

Brown algal preparations have been used as detoxifying agents.31,32 The iodine and other elements in the seaweeds inhibit absorption of similar radioactive elements by the body. In addition, there is some chelation of contaminants such as Strontium 90 by alginates in seaweeds. More recently, it has been demonstrated that
Undaria ingestion assists in eliminating dioxins in rats.33

Feeding on beach-cast seaweeds or seaweed-treated pasture is known to improve health and increase disease resistance in sheep and cattle.-34,3-5 Seaweed and other natural polysaccharides also alter the bacterial spectrum of the gut, indicating a possible mechanism for these observed effects.34,36

Antiviral Effects

Brown seaweeds, including the commonly eaten Undaria, have inhibitory effects on herpes viruses.
Herpes viruses are important human pathogens and include Herpes simplex (HSVI), genital herpes (HSVII), Varicella/chicken pox/shingles, cytomegalovirus, Epstein-Barr virus (EBV), herpes 6, 7 (Roseola, post-transplant infections), and herpes 8 (associated with Kaposi's sarcoma). In Japan, where ingestion of brown seaweed in the diet averages 2-3 g per day with a high of 12 g calculated as dry weight,37 there is a lower rate of reactivation of HSVI,38 and the lowest levels of HSVII compared to other countries.39

Acyclovir (ACV) and its derivatives comprise the most common drug group used against herpes infections. These pharmaceuticals inhibit viral DNA polymerase, thereby preventing viral replication. ACV-resistant viral strains are prevalent in 5 percent of all HSV infections in immunocompromised patients.40 Alter- natives that do not give rise to drug resistance would be invaluable.

Ingestion of Undaria led to inhibition of reactivation of herpes and amelioration of active infections in a patient study.41 In this study, one patient with an ACV-resistant HSV 11 recurrent infection experienced no symptoms for 3
months while taking an Undaria supplement.

In vitro, Undaria extracts and purified galactofucan sulfate from Undaria had inhibitory effects on forty different clinical strains of HSVI and HSVII, of which half were ACVresistant. 42 The mechanism of inhibition was via blocking the receptor on the cell surface that is normally used by the viruses to enter cells.43 In other studies, Undaria extracts have also inhibit- ed EBV, HSV, and human immunodeficiency virus (I-IIV).44-48 Undaria extracts were also shown to have additive effects with the antiretroviral drug zidovudine in an animal model.49

In the studies by Ohigashi et al.44 and Hudson et al.,46 organic sol- vent-soluble fractions of Undaria were demonstrated to have antiviral properties. However, most research attention has been directed toward the antiviral effects of anionic polysaccharides (which are water soluble).14,1-5,42,43,48

The mechanism of viral inhibition by large anionic molecules (by inhibiting viral entry to cells) does not generate resistant strains to the same degree as acyclovir-type drugsl5 (which inhibit viral replication) and the inhibition covers a wide spectrum of viral strains.14,15 If viral inhibitory polyanions, such as those found in brown seaweeds,
can supplement conventional therapies, the total amount of other drugs required and the emergence of resistant strains may be reduced.

Anticancer Effects

The lower incidence of breast cancer in the Japanese and Korean populations4,5,7 has intrigued researchers. Researchers have found that dietary brown algae and their extracts inhibit carcinogen-induced breast cancers,
lung metastases, and leukemia in animal models.6,22,26,44,48-51 Similarly, tests on the seaweed extracts
in bacterial systems revealed that the extracts had a pro- found anfimutagenic quality.52,53

Most recently Funahashi et al.6,26 have shown that wakame extracts (as wakame soaked in animals' drinking water)
have a potent inhibitory effect on the progression of mouse mammary tumors. Similar extracts produced an equally profound apoptotic effect on breast cancer cells in vitro while the extracts were non- toxic to ordinary
breast cells.

From the animal model experiments, when Undaria or other brown seaweed was included in the animals' diets,
it is very clear that there is a direct anticancer effect of ingestion although the active components have not been determined. Over the years, it has variously been attributed to iodine,54,55 tryptophan, 56 fucoidans,50 or vitamins.6

The Viral Connection with Breast Cancer

There has been a recent revival in research on viral connections with breast cancer.4,57-59 Human homologues of mouse mammary tumor virus, papilloma viruses, and herpes viruses, chiefly EBV, are implicated. The lower rates of herpes in seaweed-eating populations may be a cofactor in the observed lower rates of breast cancer.
Seaweed ingestion, which is already thought to be connected with reduced rates of breast cancer,5 may, perhaps, elicit these protective effects via inhibition of herpes viruses.

Effects on Immunity and Inflammation

Stimulation of T-cell multiplication in vitro by algal extracts59,41 may account for in vivo observations by other researchers, including increased monocytes in cattle who were fed seaweedextract sprayed grasses.35
The extensive gut lymph tissue would contact seaweeds passing through the gut. Specialized T cells in gut lymphatic tissue are important in achieving a rapid response to pathogens, in particular, to viruses such as
HSV 161,62 and may also modulate intestinal lipid metabolism. 63

Inflammatory disorders, such as psoriasis and some types of colitis, are characterized by an excessive presence of leukocytes and may be ameliorated by seaweed ingestion.
Algal-derived fucoidans inhibit the passage of leucocytes into tissues by receptor blocking.
These fucoidans are being investigated clinically for their potential to prevent destruction of postischemic heart muscle by invading leucocytes.18

Effects on Plasma Cholesterol and Hypertension

Many foods are known to reduce cholesterol levels and brown algae fall into this category.
Undaria ingestion results in lower cholesterol levels in rats.64
This effect on lipid processing seems to be the result of stimulation of liver enzymes.65 Undaria fucogalactan fractions were shown to reduce lipid clearance times dramatically when introduced intravenously. 13
The fucoidan component may block the macrophage scavenger receptor that is involved in low-density lipoprotein uptake.66

Undaria contains substantial amounts of laminine and similar tetrapeptides, which have been shown to have angiotensin converting enzyme inhibitory qualities both in vitro and in vivo.67 Ingesting 3.6 g per day of Undaria (wakame) for 4 weeks resulted in a 14 mm Hg drop in systolic blood pressure in Asian patients who had hypertension.68 Relying on on exchange properties, rather than laminine, a Swedish clinical study
i found that ngesting potassium-loaded seaweed fibers countered hypertension i sucessfully.69

Mineral Contents

Mineral concentrations vary according to the growth environment and age of marine algae.
Iodine or trace-element requirement is currently the most common reason for seaweed supplementation.
The maximum tolerated dose of 1000 jig of iodine per-day
(according to the joint Food and Agriculture Organization/World Health Organization
Expert Committee on Food Additives) can be reached with only small amounts of some kelps 1 and should be assessed carefully by practitioners. Arsenic is considered to be toxic in the form of the inorganic salt when it consumed in excess of 2 pg per kg body weight per day.70 Arsenic is also an essential trace element with a recommended minimum intake of 12-50 gg per day.71

In broad terms, annual growth algae such asUndaria contain the lowest amounts of minerals and can, therefore,
be consumed in larger quantities. For example, Tasmanian Undaria contains 53 pg per g of iodine and 0.96 pg
per g of arsenic.
* (Marine Resources data, personal communication.)

Conclusions

Brown algae as either food or in supplement form may provide useful additional therapy for treating herpetic viral infections and some cancers. Other benefits include mild antihypertensive- and cholesterol-reducing effects.
Used with caution, so as not to exceed the maximum iodine or arsenic intakes, these algae also provide valuable
mineral supplementation.

Acknowledgment

The author wishes to acknowledge the support of Marine Resources Pty. Ltd.,
Tasmania, Australia and the assistance of Julia Pearson.

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J. Helen Fitton, Ph.D., is a scientific consultant in Hobart, Tasmania, Australia.

Corresponding author.

1 Anubha Mountain Health Retreat. 680 Summerleas Road, Kingston, Tasmania, Australia.

2 Novost Pty Ltd, 53 Channel Highway, Kingston, Tasmania 7050, Australia.

3 Suite 105, 86 Murray Street, Hobart Tasmania.

4 Marine Biomedical Research Level 10, 39 Murray Street, Hobart, Tasmania 7000, Australia

5 Healing World 124, 16th Avenue Tauranga New Zealand.

6 Department of Pathology, University of Chicago, USA.

Abstract

Background We sought to assess whether GFS, a proprietary preparation of Tasmanian Undaria pinnatifida has
effects on healing or re emergence of Herpetic infections, and additionally, to assess effects of GFS in vitro.
Undaria is the most commonly eaten seaweed in Japan , and contains sulphated polyanions and other components with potential anti-viral activity. Herpes simplex virus type 1 (HSV-1) infections have lower reactivation rates and
Herpes type 2 (HSV-2) infections have lower incidence in Japan than in the west.

Methods Patients with active (15 subjects) or latent (6 subjects) Herpetic infections (HSV-1,2, EBV, Zoster) were
monitored for response to ingestion of GFS. GFS extract was tested in vitro for human T cell mitogenicity and anti-Herpes activity.

Results Ingestion of GFS was associated with increased healing rates in patients with active infections. In addition, patients with latent infection remained asymptomatic whilst ingestingGFS. GFS extract inhibited Herpes viruses in vitro and was mitogenic to human T cells in vitro.

Conclusions Ingestion of GFS has inhibitory effects on reactivation and is associated with increased rate of healing after Herpetic outbreaks. GFS extract potently inhibited Herpes virus in vitro , and had mitogenic effects on human T cells.

Background

Herpes viruses are important human pathogens, causing both primary and secondary infections that range from trivial mucosal ulcers to life threatening disorders in immunocompromised patients (1,2). The Herpes group includes HSV-1, HSV-2, Herpes Zoster (chicken pox/shingles), HCMV (human cytomegalovirus), EBV, Herpes 6, 7
(Roseola, post transplant infections) and Herpes 8 (associated with Kaposi sarcoma).
The conventional treatment of these infections is with drugs such as acyclovir (ACV) that target the viral DNA
polymerase. Whilst these drugs are undoubtedly efficient, their long-term use has led to the development of
resistant viral strains that now comprise 5% of all HSV infections in immuno-compromised patients (1).
Finding non-toxic alternatives to these drugs is of interest to patients and to drug companies investigating more effective remedies. Whilst the epidemiology and serotype of Herpes infections varies widely within the world community, as a result of differing subpopulations and childhood exposure rates, it seems that the
incidence of HSV2 in Japanese women is substantially less than that of American women.
(e.g., 50% seroprevalence in one study of African-American women vs. 7% in Japan) (3).

We also noted that the reactivation of Herpes simplex I (HSV-1) as measured by shedding (4) is considerably less
in the Japanese population than in the west. However, EBV, some strains of which are associated with malignancies, affects over 90% of the population world wide (5), and seropositivity for CMV is higher in the
Japanese population (6).

Several plants have been shown to have anti-viral activity, but edible brown marine algae are unique in that they
are a regular part of the Japanese daily diet. Key anti-viral active agents within brown algae are the sulphated
polyanions (7,8,9). In recent years both natural and synthetic sulphated polyanions have been a research focus
for their potential therapeutic activity.
They are considered to act principally by inhibiting the entry of coated viruses such as Herpes into host cells
(7,8,9), by competing for receptors at the cell surface. This activity is, in part, due to the similarities of the algal polyanions to mammalian heparin sulphate molecules.
There are a number of receptors (including a heparin sulphate receptor) expressed differentially on various cell types, which provide entry points for Herpes viruses (10). This mechanism of inhibition contrasts with that of commonly used drugs such as ACV which act as nucleic acid inhibitors, preventing viral replication after the virus has entered the cell.

The main seaweed in the Japanese daily diet is Undaria pinnatifida, commonly known as ¡®wakame¡¯ (11).
The major sulphated polyanion present in Undaria is characterised as a ¡®galactofucan sulphate¡¯ (12,13).
Other sulphated polyanions such as the synthetic dextran sulphates, pentosan sulphates, clinically used heparins, and seaweed-derived carageenans have all been investigated for anti-viral activity in humans, either systemically
or topically (7,8,9). However, to date, Undaria ingestion has not been clinically assessed in a western population
for its effects on common coated viruses such as HSV-1 and 2 which have reduced incidence in the Japanese population (3,4). Undaria extracts have been shown to have anti-viral effects on HSV-1 in vitro (14) and also against the Herpes group virus Epstein Barr Virus or EBV (15). Specific molecules were not identified in these studies, thus the observed anti-viral activity may arise from either or both polyanions and other components.
Undaria aqueous extract (made by boiling Undaria in water) and a partially purified galactofucan sulphate extract
were recently assessed for activity against clinical isolates of Herpes I and Herpes II in vitro (16,17).
Twenty of the viral strains were sensitive to ACV, and nineteen were resistant. In the test, both types of extract were active against all strains, and more active against strains of HSV-2 than strains of HSV-1.
The mode of action was shown to be blocking attachment and entry of HSV into the host cells.

Undaria fractions have also been shown to have immune stimulating qualities in vitro (18)
and other brown seaweed fractions have been shown to have immunological effects in vivo
(19,8,9).

The mechanism of viral inhibition by polyanions such as galactofucan does not generate resistant strains at the
same rates as ACV (7). Aside from the inhibition of viral infectivity, it is possible that immune stimulation via
polyanions or other components, increases anti-viral activity (18,19)

We hypothesised that Undaria ingestion may be associated with resolution of Herpes infections in a western population, and that this would be reflected by inhibition of HSV infectivity and increased T cell activity in vitro.
In this study, a proprietary Undaria preparation ¡®GFS¡¯ was assessed for effects on the healing of,
and the inhibition of outbreaks of HSV infections in otherwise healthy patients. In addition,
whole aqueous extracts of the GFS were assessed for effects on HSV infectivity in vitro and on human T cell stimulation in vitro.

Materials and methods

A) GFS
GFS was prepared from Tasmanian Undaria pinnatifida and supplied in 560 mg capsules by Marine Resources
Pty Ltd. It comprises solely of the alga, processed to complete dryness in the dark within a day of harvest,
in hygienic conditions. Batches were milled and mixed to ensure uniformity.

B) Patient study
Seventeen patients were recruited for the study by health practitioners. Patients gave verbal informed consent to
the study. Health practitioners monitored the patients¡¯ health.
There are no known adverse effects related to the ingestion of Undaria. No other anti-viral medications
were taken at the same time as GFS. The duration of the study was from one month to 24 months.
Patient ages were from less than 10 years up to 72 years.

In total, seven cases of HSV-1, five cases of HSV-2, three of active Herpes zoster (one chicken pox, two shingles)
and two of EBV were assessed. Results are presented in tables 1 and 2. Patients suffering from HSVI typically experienced lesions lasting ten days. Patients with HSVII had experienced recurrent events of varying severity, sometimes overlapping, of up to ten days in duration. One patient with shingles reported ¡®cycle¡¯ duration of
over ten days. Others did not report on duration.

Dosage levels were chosen to fall within the limits of normal dietary seaweed ingestion in Japan .
Fifteen patients with active Herpetic viral infections were given four 560 mg capsules of GFS per day for ten days as
a ¡®therapeutic dose¡¯. All patients except subject 14, table 1 (primary zoster infection) were suffering repeat outbreaks of known aetiology

Six patients with latent HSV-1 or 2 were given two capsules per day as a ¡®maintenance dose¡¯.
Four of these patients, identified by an asterisk *, were continuations of treatment after the active infection phase.
One patient (subject 3, table 2) took four capsules per day.

C) In vitro effects on HSV
GFS was mixed 1:40 w/v with distilled water and boiled for 5 minutes. The liquid was filtered through a 0.45¥ìM
filter for sterilization and stored at . 20 degrees Celsius. An aliquot of the preparation was dried and the weight
was obtained to determine the concentration.
The concentration used was the dry weight of the dissolved solids present.

Immortalized human fibroblasts, HF cells, were grown in Minimal Essential Media supplemented with glutamine, antibiotics, and 10% foetal bovine serum (FBS). Maintenance medium was supplemented with 1% FBS.
Laboratory strains of HSV and HCMV (American type culture collection) were tested in this study.
A stock of each virus was grown in cultured HF cells and aliquots were frozen at . 70oC.
The titre of each virus was determined by a plaque assay using HF cells in 24-well plates with an agarose overlay.

D) T cell stimulation in vitro
T cell mitogenicity was evaluated by chromium uptake. Whole T cell preparations were obtained from buffy coats
from pooled human blood samples.
They were incubated in RPMI supplemented with 10% heat inactivated foetal calf serum,
5mM L-glutamine, 5X10 .5 M 2- mercaptoethanol and 30U/ml gentamycin. Incubation for 72 hours was at 5%CO2,
37oC in 24 well plates. Cells were incubated with either GFS whole extract (at 25, 125 and 250 mcg/ml as 1%, 5%
or 10% of total culture volume prepared from a stock solution at 2.5mg/ml) or with the known mitogens (PHA) (1mcg/ml) or Concanavalin A (ConA) (1mcg/ml). Each concentration was assessed in triplicate (n=3)

Results

Patient study.
Results are presented in tables 1 and 2.

Active infection- table 1
All fifteen patients with active Herpetic viral infections experienced significant lessening or disappearance of symptoms. No adverse side effects were noted during the study.

Two patients (subjects 4 and 5, table 1) with noncompliant dosage regimes resolved infections in normal time,
but noted no spread of lesions (as occurred during previous outbreaks).
Reduction in lesion severity and rapid clearance were noted in two patients (subjects 6,7, table 1), and pain reduction as compared to previous events was noted by two patients (subjects 2,14, table 1). Two females with genital HSV-2 had persistent lesions which resolved during the course of treatment (subjects 8,10, table 1).

In two cases of diagnosed EBV, one clear at four and the other by ten days. In the latter patient a chronic sinus condition also cleared (subjects 11,12, table 1)

Over ten days, faster drying of zoster lesions and increased speed of normal cycle as compared to previous
outbreaks was noted by a male patient (subject 15, table 1) although no reduction in pain was reported.
In an adult male suffering primary zoster (chicken pox) lesions of whole body (subject 14, table 1),
pain reduction and rapid healing of lesions were noted.
An elderly female patient noted a reduction in pain and visible skin lesions during the ten day course,
and had no recurrence of symptoms whilst on a lower dose for the next two months.

Latent infections .table 2
All six patients on maintenance doses noted inhibition of further outbreaks of infection.
No adverse side effects were noted during the study.

HSV-1 outbreaks were inhibited in two patients taking a maintenance dose over three months and two years respectively (subjects 1 and 2 in table 2). Low grade HSV-1 associated keratoconjunctivitis in the former patient
was also inhibited

GFS ingestion correlated with inhibition of a previously persistent HSV-2 infection for three months in subject 4,
table 2. In this patient, the infection was ACV resistant and outbreaks had been apparent on a two weekly basis
for over a year.

HSV-2 outbreaks at the genital site were inhibited in two other female patients whilst taking a maintenance dose of two capsules per day, for one month (subjects 5,6 , table 2).

Low grade recurrent Herpes zoster (shingles) lesions of the torso were inhibited for two months in an elderly
patient whilst maintaining a dose of four capsules per day (subject 3, table 2).

C) In vitro effects on HSV
Herpes viruses were assessed for infectivity of human fibroblasts cells in vitro.
Inhibition by GFS extract was noted as shown in table 3.

D) T cell stimulation in vitro
GFS extract was assessed for effects on whole human T cell preparation in vitro.
After incubation with GFS extract or mitogens PHA and ConA, for 72 hours the relative uptake of chromium was assessed as a measure of mitogenicity.
The lowest concentration of GFS extract tested (25mcg/ml) exerted a four fold mitogenic effect on T cells,
over 50% of the mitogenic potency of the known mitogens PHA (six fold) and ConA (seven fold).
Paradoxically, increased concentrations of the whole extract showed decreasing effects on mitogenic activity.
This may be accounted for by the increasing physical inhibition due to increased viscosity in the culture media,
or the increasing concentration of unidentified inhibitory components present in the extract.
Results are shown in Figure 1. Additional studies illustrated little effect on NK cell activity and no effects on L929
fibroblast growth over 24 or 72 hours (results not shown).
There was no bacterial contamination of the GFS (results not shown), thus the presence of bacterial lipopolysaccharides (which may also act as mitogens) was ruled out.

Discussion

This study was carried out to assess the effects of GFS in patient studies and in vitro.

GFS was ingested by patients suffering active or latent Herpes infections. Results indicated firstly,
a positive effect on healing, and secondly, inhibition of outbreaks in cases of HSV-1, HSV-2, ACV resistant
HSV-2, and zoster. There were no adverse side effects noted, and GFS was well tolerated by all subjects.
Reduced pain levels were noted in some cases.

A particularly noteworthy result in this study was inhibition of an ACV resistant case of
HSV-2. HSV-2 is a sexually transmitted disease of increasing incidence (4).
In part, this is due to the fact that partner transmission may occur during asymptomatic shedding (4) or unrecognised minor outbreaks. Suppressive therapies such as ACV have been tested for their ability to inhibit
shedding (20). However, for long-term use, non-toxic alternatives such as GFS may be preferred by patients,
who perceive long-term conventional drug use as detrimental. In addition, GFS may reduce the
generation of resistant strains which arise through prolonged use of drugs such as ACV.

GFS extract potently inhibited HSV and HCMV infection of human cells, and stimulated human T cell proliferation
in vitro. In other studies (16, 17) extracts were active against clinical strains of HSV.
These mechanisms may be relevant to the observed clinical response.

Human T cell proliferation in vitro observed here concurs with observations by Shan et al (18).
Shan found that in the presence of various seaweed extracts including Undaria,
lymphocyte proliferation and cytotoxic T cell activity was enhanced, but that NK cell activity was not.
Cytotoxic T cells play a prominent role in the immune response in the local environment of HSV-2 infections (21),
and have been shown to be key elements of immune defence against HSV-I and other viruses (22,23).
Thus the observed T cell mitogenic effect caused by GFS in vitro may be related to enhanced immunity in vivo.

Sulphated molecules with similarities to sulphated galactofucan, such as dermatan sulphate have been shown to
be present in plasma after oral ingestion (24) Macromolecules are known to pass through the gut into plasma
and into lymph via the extensive gut lymphoid system (25) where they may affect immune function
The gut lymphoid system or ¡®GALT¡¯ comprises Peyers patches and gut cryptopatches where different types of
T cells and other immune cells participate in uptake or contact with gut contents.
T cell uptake or contact with GFS by either Peyers patch or gut cryptopatch dwelling cells may result in the proliferative response noted here in vitro.
However, immune indicators were not measured in this study.

Whilst further research is called for, the results obtained in this study show that ingestion of GFS is associated
with resolution, reduced pain and outbreak inhibition of Herpes virus infections. Studies regarding uptake, immunostimulation and anti-viral effects of GFS are in progress.

Conclusions

The rise in nucleic acid inhibitor resistant strains of HSV-2, and the need for less aggressive anti-viral therapies, indicates a role for alternative therapeutic routes such as that demonstrated here for GFS.

Ingestion of GFS has inhibitory effects on reactivation of HSV and positively affects the resolution of active Herpes
infections. GFS has inhibitory effects on Herpes infectivity in vitro and a T cell mitogenic effect in vitro.

The mechanism of inhibition of Herpes infections may be, in part, mediated by galactofucan present in the GFS.

Acknowledgements

Marine Resources Pty Ltd supplied GFS for this study. Thanks to C Trambas (University of Tasmania) for data on
T cell mitogenicity.

Declaration of competing interests

Russell Cooper, Ken Thompson, John Godwin, Kate Elliot have no declared interests.
JH Fitton and Charles Dragar are employed Marine Biomedical Research Tasmania Australia.

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Figure legend

Figure 1. T cell mitogenicity in the presence of GFS whole extract in vitro.

GFS extract assessed at 25, 125, 250micrograms/ml (as1%, 5% and 10% of culture medium
from a stock solution at 2.5mg/ml). PHA and ConA were assessed at 1mcg/ml. N=3 in all
cases. Blue bars indicate average readings, red bars indicate standard deviations.

Table 1. Patients with active Herpes infections

Patient	Sex	Age	Virus	Site infection	Resolution of infection?	If on maintenance, Inhibition of outbreaks	Comments
1	M	50	HSVI	Orolabial	Yes, no progression to lesion	Yes, inhibition of further outbreaks on	Varied dosage, consistent inhibition.
2	F	14	HSVI	Orolabial	Yes, very severe outbreak resolved within course.	N/a	Patient noted rapid reduction in pain
3	F	72	HSVI prodro -me	Orolabial (prodrome) and ocular conjunctiva	Yes, no progression to lesion	Yes, continued Inhibition of low grade conjunctival HSVI for three months	Notes improvement in skin condition.
4	M	40	HSVI prodro -me	Orolabial	Yes, in normal time.	N/a	Not taken consistently. No benefit noted but no spread of lesion.
5	F	50	HSVI active lesion	Orolabial	Yes,in normal time	N/a	No spread of lesion and pain reduced. Took half dose only.
6	F	47	HSVI	Orolabial	Yes, reduction inlesion severity	N/a	No recurrence, no spread of lesion.
7	F	47	HSVI	Orolabial	Yes, rapid clearance compared to previous.	N/a	Post chemotherapy outbreak (breast cancer)
8	F	20	HSVII	Genital	Yes, lesions cleared.	N/a
9	F	42	HSVII	Genital	Yes. Existing lesion healed.	Yes, inhibition of further outbreaks on maintenance dose 3 mths.	Prior two weekly outbreaks of ACV resistant strain of HSVII.
10	F	23	HSVII	Genital	Yes, chronic lesion healed	N/a
11	F	17	EBV	systemic	Yes	N/a	Normal blood exam after 4 days course.
12	F	<10	EBV	Systemic	Yes, EBV symptoms absent at ten days	N/a	Three capsules per day Chronic sinus infection also cleared
13	F	85	Zoster (Shang les)	Torso	Yes	Yes, inhibition for two months.	Relief from lesions at 4 capsules per day
14	M	Adult	Zoster (Chick en Pox)	Whole body sores	Yes	N/a	Pain reduction, rapid clearing of lesions.
15	M	40	Zoster (Shang les)	T7,8, dermatome Right side	Yes	N/a	Faster drying of lesions, increased speed of cycle, no change in pain

Table 2. Patients with latent Herpes infections.

patient	sex	age	virus	Site of infection	Also treated for active infection?	Inhibition of outbreak whilst on maintenance dose?	Comments
1	M* (1)	50	HSVI	Orolabial	Yes Existing lesion healed.	Yes, inhibition of further outbreaks on maintenance dose >2 years.	Varied dosage, consistent inhibition.
2	F* (3)	72	HSVI prodrome	Orolabial (prodrome) and ocular conjunctiva	Yes, no progression to lesion	Yes, continued Inhibition of low grade conjunctival HSVI for three months	Notes improvement in skin condition.
3	F* (13)	85	Zoster (shingles)	Torso	Yes	Yes, inhibition for two months.	Relief from lesions requires 4 capsules per day
4	F* (9)	42	HSVII	Genital	Yes. Existing lesion healed.	Yes, inhibition of further outbreaks on maintenance dose 3 months.	Prior two weekly outbreaks of ACV resistant strain of HSVII.
5	F	41	HSVII	Genital	No(not taken during active infection)	Yes, inhibition on two capsules per day for 1 month.	Did not take during active lesion outbreak.
6	F	36	HSVII	Genital	No(not taken during active infection)	Yes, inhibition on two capsules per day for 1 month.	Did not take during active lesion outbreak.

* the number in brackets refers to the number assigned to the same patient in table 1.

Table 3. IC50 for GFS whole extract as measured by infectivity of HSVI, HSV2 and HCMV
(human cytomegalovirus) in human fibroblasts.

Herpes virus	GFS 1:40 w/v
HSV - 1, strain F	3.1 ug/ml
HSV - 2, strain G	1.6 ug/ml
HCMV, AD169	2.5 ug/ml
HCMV, D16	2.5 ug/ml