One such casualty of the drug approval process is a red marine alga in the
family of Dumontiaceae. Research on antiviral carbohydrates from marine red
algae indicate a high potential for low-cost, broad spectrum antiviral agents.
Further research in the family of Dumontiaceae produced two patents where
clinical efficacy for herpes I and II was clearly shown. The treatment was
effective for treating subjects (e.g. human patients) both prior to and
subsequent to herpes infection. It was used topically to alleviate symptoms
associated with herpes infections or preferably systemic, by oral
administration, to eradicate the virus and thereby prevent symptom recurrence.
No side effects or toxicity were noted. This treatment, which now must be
considered alternative, suggests a breakthrough in the discovery of natural
immunomodulatory and antiviral agents.
Recent research and gathering of anecdotal evidence on the health benefits
and antiherpetic action of the red marine alga, Dumontiaceae, has yielded much
promise. Its use as a topical has been further documented and thought superior
to acyclovir. It was shown to be clinically effective against herpes zoster
infections as well. Anecdotal reports from patients suffering from Epstein
Barr (another herpes virus) and Candida have shown marked improvement in a
short period of time through oral administration (systemic).
Microalgal Ointment for Treating Herpes
October 22, 2000
Microalgal Ointment for Treating Herpes
Developed at Ben-Gurion University of the Negev
An efficient antiherpes ointment containing material from microalgae has been
developed by researchers at Ben-Gurion University of the Negev. The natural
product, found to be effective against various herpes infections, shortens the
duration of the disease, prevents its spread, and significantly reduces pain.
The new material is being registered as a commercial patent.
The medicine contains large quantities of special polysaccharides, unique
large sugar molecules produced by the alga. Herpes viruses commonly cause open
wounds on skin, particularly on the face and reproductive system, but they
also cause of shingles, a painful condition often appearing on the back. It
has been found that the algal polysaccharides prevent further spreading of the
virus, thus greatly easing the pain of herpes sufferers.
Researchers working on the development of the product include Prof. Shoshana
Arad, Director of the Institute for Applied Biosciences at Ben-Gurion
University of the Negev, and Dr. Mahmoud Huleihel, who carried out the
research during his post-doctoral work. Tests performed at Hadassah Medical
Center in Jerusalem and at the Pasteur Institute in Paris have confirmed the
preliminary results pointing to the benefits of the ointment.
Current herpes� treatment is by acyclovir, a synthetic drug that is thought
to block transcription of the viral DNA. Because this drug interferes with the
processing of cellular DNA, the body may be exposed to negative side effects.
The new formulation developed at BGU is significantly less toxic and much more
efficient than the drug. Since it is a natural product, unpleasant side
effects are less likely, even with long-term use. Additionally, acyclovir-resistant
herpes were effectively treated with the polysaccharides.
According to Prof. Shoshana Arad, the results of the research are highly
promising. Initial tests on the product, primarily on herpes sufferers, have
already proven its great efficacy. Aside from her work on medical applications
of microalgae, Prof. Arad has begun working with Estee Lauder Companies on the
development of a new cosmetic product line also based on microalgae.
Prof. Arad is very optimistic about the future of building industries in the
Negev based on microalgae:
�These organisms, which usually live in salty water, could be the basis for
unlimited commercial industrial applications. Our dream is to establish an
industrial park to produce natural products based on microalgae, including
health foods, cosmetics and medicines,� declares Arad.
An agreement recently signed with Frutarom Company for the development of
algae-based products will enable strategic collaboration with pharmaceutical,
cosmetic, and health food companies, for which Frutarom will manufacture the
goods. Development work has already started on the new antiherpes ointment,
with the initial test results having attracted international interest.
For further details, contact Prof. Shoshana Arad: Tel: (office) 07-646-1963/4;
(mobile) 058-816-421.
Or Amir Rozenblit, BGU Spokesperson: Tel: 972-7-6461802 / 972-7-6477717/6;
Fax: 972-7-6472803; E-mail: rosenbli@bgumail.bgu.ac.il
Calcium
Spirulan, an inhibitor of enveloped virus replication, from a blue-green
alga Spirulina.
by Hayashi et al. 1996. Pub. in Journal of Natural Products, 59, 83-87.
Japan.
Bioactivity-directed
fractionation of a hot H2O extract from a blue-green-alga Spirulina
platensis led to the isolation of a novel sulfated polysaccharide named
Calcium Spirulan (Ca-SP) as an antiviral principle. This polysaccharide was
composed of rhamnose, ribose, mannose, fructose, galactose, xylose, glucose,
glucuronic acid, galacturonic acid, sulfate and calcium. Ca-SP was found to
inhibit the replication of several enveloped viruses, including Herpes
simplex virus type 1, human cytomegalovirus, measles virus, mumps virus,
influenza A virus and HIV-1. It was revealed that Ca-SP selectively
inhibited the penetration of virus into host cells. Retention of molecular
conformation by chelation of calcium ion with sulfate groups was suggested
to be indispensible to its antiviral effect.
An extract from
spirulina is a selective inhibitor of herpes simplex virus Type 1
Penetration into HeLa Cells.
by Hayashi et al. 1993. Pub. in Phytotherapy Research, Vol. 7. 76-80. Japan.
The water soluble
extract of spirulina achieved a dose-dependent inhibition of the replication
of herpes simplex virus type 1 (HSV-1) in HeLa cells within the
concentration range of 0.08-50 mg/mL. This extract proved to have no
virucidal activity and did not interfere with adsorption to host cells.
However, the extract affected viral penetration in a dose-dependent manner.
At 1 mg/ml the extract was found to inhibit virus-specific protein synthesis
without suppressing host cell protein synthesis if added to the cells 3
hours before hamsters at doses of 100 and 500 mg/kg per day.
Effects of
polysaccharide and phycocyanin from spirulina on peripheral blood and
hematopoietic system of bone marrow in mice.
by Zhang Cheng-Wu, et al.. April 1994. Nanjing Univ. China. Pub. in Proc. of
Second Asia Pacific Conf. on Algal Biotech. Univ. of Malaysia. China.
C-phycocyanin and
polysaccharide were isolated and purified from spirulina. By using the
techniques of colony forming unit-erythroid (CFU-E) culture of fetal liver
cells in mice in vitro, C-phycocyanin was found to possess high
erythropoietin (EPO) activity. The effects of polysaccharide and phycocyanin
on the periphyeral blood and bone marrow hematopoietic stem and progentitor
cell in normal, irradiated and anemic mice were studied. These studies
demonstrate the unique capacity of C-phycocyanin and polysaccharide to
influence the differentation and proliferation of committed hematopoietic
progenitor cell. Stimulated recovery by C-phycocyanin and polysaccharide was
observed in leukocyte and bone marrow nucleated cell counts and the number
of CFU-GM colony formation in mice after single whole-body gamma-ray
irradiation. The C-phycocyanin and polysaccharide can lower the anemic
degree of mice combined with treatment of gamma-ray irradiation and
benzohydrazine hydrochloric acid peritonerl injection.
Enhancement of
antibody production in mice by dietary spirulina.
by Hayashi, et al. June 1994. Kagawa Nutrition Univ. Japan. Pub. in Journal
of Nutr. Science and Vitaminology. Japan.
Mice fed a
spirulina diet showed increased numbers of splenic antibody- producing cells
in the primary immune response to sheep red blood cells (SRBC). However,
immunoglobulin G (IgG) - antibody production in the secondary immune
response was hardly affected. The percentage of phagocytic cells in
peritoneal macrophages from the mice fed spirulina diet, as well as the
proliferatiom of spleen cells, was significantly increased. Addition of a
hot-water extract of spirulina (SHW) to an in vitro culture of spleen cells
markedly increased proliferation of these cells, whereas culture of thymus
cells was scarcely affected. The spirulina extract also significantly
enhanced interleukin-1 (IL-1) production from peritoneal macrophages.
Addition to the in vitro spleen cell culture of SHW as well as the
supernatant of macrophages stimulated with SHW resulted in enhancement of
antibody production, that is, an increase of the number of PFC. These
results suggest that spirulina enhances the immune response, particularly
the primary response, by stimulating macrophage functions, phagocytosis, and
IL-1 production.
Immune
enhancement potential of spirulina in chickens.
by M. Quereshi, et al. August 1994. Poultry Science Assoc. Dept. of Poultry
Science, North Carolina State, NC. Pub. in Journal of Poultry Science Vol
73, S1. p.46. USA.
Effects of
spirulina on the immune function of chickens were examined. Macrophage
cultures treated with a water soluble extract of spirulina exhibited
enhanced phagocytosis and induced tumorcidal factor secretion. In the second
study, 0, 10, 100 and 10,000ppm spirulina in corn/soy diets were fed to
Leghorn chickens. Larger thyus glands, higher NK activity and CBH response
were observed in the 10,000ppm spirulina treated chickens. Percent
phagocytic macrophages and secondary antibody response were also greater
than non-treated chickens. The data suggests spirulina exposure improves
immune performance of chickens without adversely affecting other performance
characteristics.
Immunomodulary
effects of spirulina supplementation in chickens.
by M. Qureshi, et al. May 1995. North Carolina State. Pub. in Proc. of 44th
Western Poultry Disease Conference, pp 117-120. USA.
Chicken
macrophages exposed to a water-soluble spirulina extract show enhanced
phagocytic activity in vitro suggesting activation of mononuclear phagocytic
system function. Furthermore, dietary supplementation of Spirulina (1,000
ppm to 10,000 ppm) improved thymic weights, enhanced CBH response, increased
tumor cell killing by NK cells and doubled the macrophage phagocytic
potential over chickens fed a basal diet. Chicks on 1,000 ppm Spirulina diet
cleared significantly more E. Coli from circulation at 30 and 40 minutes
post i.v. inoculation. Similiar reduction was seen in clearing bacteria from
spleen after 80 minutes post i.v. inoculation of Staph. Aureus in chicks fed
10,000 ppm over controls. Chicks in groups from 1,000 ppm to 16,000 ppm
Spirulina treatments showed enhanced CBH response. Chicks in 1,000 ppm group
exhibited enhanced E. Coli clearance between 30 to 60 minutes and decreased
splenic bacterial counts at 80 minutes post inoculation. These studies imply
Spirulina enhances several immunological end points in chickens both during
in vitro and in in vivo exposures.
Immunostimulating
activity of lipopolysaccharides from blue-green algae.
by L. Besednova, et al. 1979. Pub. in Zhurnal Mikrobiologii, Epidemiologii,
Immunobiologii, 56(12) pp 75-79. Russia.
The whole cells
of blue-green algae and lipopolysaccharides (LPS) isolated from these cells
were shown to stimulate the production of macro- and microglobulin
anti-bodies in rabbits. The macro- and microphage indices in rabbits
increased significantly after the injection of LPS isolated from blue-green
algae 24-48 hours before injecting the animals with a virulent Y.
pseudotuberculosis strain. Besides, the inhibiting action of this strain on
the migration of phagocytes to the site of infection was abolished
immediately after the injection. The use of the indirect hemaglutination
test allowed to provide the absence of close antigenic interrelations
between blue-green algae and the following organisms: Spirulina platensis,
Microcystis aeruginosa, Phormidium africanum and P. uniccinatum. <
Is
There Any Protection?
Dumontiaceae (Cryttosiphonia woodii) is a very rare red marine alga
containing natural antiviral and immunomodulatory agents. Research
is still underway, but we know that Dumontiaceae (doo-mont-ee-ay-see-uh)
significantly interferes with the infectivity of Herpes simplex I
and II. The major questions remaining are:
1. What, in Dumontiaceae, delivers its benefits? (It is thought
the agents may be sulfated polysaccharides.)
2. Does Dumontiaceae just suppress or also kill Herpes viruses?
What
We Know
Years of study show that Dumontiaceae suppresses Herpes viruses,
especially in initial stages of an attack. Use of supplemental
Dumontiaceae increases the period of time between attacks and can
moderate or alleviate symptoms. Researchers know that Dumontiaceae
promotes a strong cell-mediated immune response to the virus. The
Herpes virus safely hides and lies dormant between attacks within
host nerve cells. If antiviral substances in Dumontiaceae can get to
them while they are napping, and either inhibit their ability to
make more of themselves or kill them outright, then Dumontiaceae may
prove to be a profound food for the control of Herpes.
We also know
that sulfated polysaccharides in Dumontiaceae work against a
critical step in viral replication both outside and inside infected
cells.
The
Details
Sulfated polysaccharides seem to inhibit viral reverse transcriptase
enzyme. This enzyme is used by the virus to direct the DNA of the
host cell to make more copies of the virus. Such parasitic action
leads a virus to replicate itself using resources of the host cell.
Inhibiting the enzyme effectively inhibits the spread of the virus
by prohibiting future generations. If sulfated polysaccharides from
Dumontiaceae interfere with viral reverse transcriptase messengers,
it is possible that even viruses responsible for the initial
invasion may be held in check and later destroyed.
Anti-viral activity of red microalgal
polysaccharides against retroviruses Marina M Talyshinsky,
Yelena Y Souprun
and Mahmoud M Huleihel
The Institute for Applied Biosciences,
Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva, Israel
� 2002 Talyshinsky et al; licensee
BioMed Central Ltd. This article is published in Open Access: verbatim
copying and redistribution of this article are permitted in all media
for any non-commercial purpose, provided this notice is preserved along
with the article's original URL.
Keywords: Red microalgae, polysaccharides, malignant cell
transformation, retroviruses, antiviral activity
Red microalgal polysaccharides significantly inhibited
the production of retroviruses (murine leukemia virus- MuLV) and cell
transformation by murine sarcoma virus(MuSV-124) in cell culture. The
most effective inhibitory effect of these polysaccharides against both
cell transformation and virus production was obtained when the
polysaccharide was added 2 h before or at the time of infection.
Although, addition of the polysaccharide post-infection significantly
reduced the number of transformed cells, but its effect was less marked
than that obtained when the polysaccharide was added before or at the
time of infection.The finding that the inhibition of cell transformation
by MuSV-124 was reversible after removal of the polysaccharide suggested
that microalgal polysaccharides inhibited a late step after provirus
integration into the host genome. In conclusion, our findings could
support the possibility that the polysaccharide may affect early steps
in the virus replication cycle, such as virus absorption into the host
cells, in addition to its effect on a late step after provirus
integration.
Retroviruses [1-3]
� viruses that contain reverse transcriptase, an RNA-directed DNA
polymerase [4,5]
� have been implicated in various types of human and animal leukemia's
and other tumors. Although there are many compounds that exhibit potent
anti-viral, and possibly anti-tumor, activity in cell culture and in
experimental animals, only very few synthetic compounds and one natural
product � alpha interferon � have so far been approved for treatment
of viral infections in man. Alpha interferon has been approved for
treatment of hairy cell leukemia, of Karposi's sarcoma and of genital
warts caused by papiloma virus [6].
A class of natural products with low mammalian toxicity
that are currently regarded as having antitumor activity are
polysaccharides of biological origin, e.g., polysaccharides from yeasts,
algae, bacteria, higher plants and fungi [7-9].
Of interest in this context are polysaccharides produced by some species
of red algae; these compounds have shown promising activity against a
variety of animal viruses [10-13].
In general, polysaccharides exhibiting antiviral potential are highly
sulfated [10,14-17].
Dextran sulfate and polysaccharides from marine algae, for example, have
been found to be potent in vitro inhibitors of HIV types 1 and 2 [15,18-20].
They inhibit HIV-1-induced cytopathogenicity and HIV-1 antigen
expression [13,18,21,22].
These sulfated polysaccharides also inhibit the activity of purified
reverse transcriptase and RNase H, which are essential for retrovirus
replication [18,20].
Some previous studies showed that algal polysaccharides exert their
inhibitory action at a very early stage (adsorption, fusion or
penetration), in the viral infection cycle [9,18,20,23-25],
whereas others showed that these polysaccharides did not interfere with
virus attachment or penetration, but they did prevent viral protein
synthesis [11,26,27].
Our previous research has shown potent antiviral
activity of a highly sulfated polysaccharide extracted from red
microalga. The polysaccharide, which consists mainly of xylose, glucose
and galactose [28], exhibits
antiviral activity against various members of the herpes family of
viruses [29]. In the current
study, the activity of this red microalgal polysaccharide against the
replication and the transforming ability of the retroviruses, Moloney
murine sarcoma virus (MuSV) and Moloney murine leukemia virus (MuLV),
was studied.
Figure 2 Kinetics of the development
of MuSV-malignant cell transformation in the presence of Porphyridium
sp. polysaccharide, dextran sulfate or carragenaan
Figure 3 Effect of polysaccharides on
cell proliferation of NIH/3T3 cells
Figure 4 Effect of time of Porphyridium
sp. polysaccharide addition on MuLV progeny release
Tables
Table 1
Inhibiting effect of red
microalgal polysaccharides on formation of foci by MuSV-124 and
on virus production, as measured by reverse transcriptase (RT)
activity
Table 2
Effect of dosage of Porphyridium
sp. polysaccharide on NIH/3T3 cell transformation by MuSV-124
and MuSV/MuLV
Table 3
Effect of time of addition of Porphyridium
sp. polysacharide on NIH/3T3 cell transformation
Table 4
Focus formation after removal
of Porphyridium sp. polysaccharide
Characterization of cell
transformation by MuSV-124 and MuSV/MuLV
NIH/3T3 cells grown in plastic dishes in RPMI medium
with 2% NBCS appear as flat cells (Fig 1a);
these cells are completely unable to grow in agar. When these cells were
infected with an appropriate dilution of MuSV-124, tiny foci of
transformed cells, with a highly refractile spindle shape, growing
randomly in a criss-cross fashion, could be detected by microscopic
observation within five days of infection (Fig. 1b).
Later, these foci gradually increased in size and compactness until they
became visible to the naked eye on day 12 to 14 after infection with
MuSV-124. The number of foci remained unchanged in these cultures during
the entire culture period. When the cells were infected with high titer
of MuSV-124 (1 ffu/cell), most of the cells were transformed two-three
days after infection. In MuSV/MuLV-infected cultures, the number of foci
increased continuously, and at any time foci of various sizes (tiny to
large) could be detected. Moreover, if these cultures were maintained
for a sufficiently long time, all the cells eventually became
transformed. Examination of the culture medium for the presence of viral
reverse transcriptase revealed that MuSV/MuLV infection yielded
virus-producing cells, whereas MuSV-124 infection resulted in the
formation of transformed cells not producing virus. It is therefore
likely that the increasing number of foci in the productively infected
cells resulted from multiple secondary infections by the virus
progenies, the smaller foci being formed by these infections. It was
found that the later the infection, the smaller the foci. Both MuSV-124-
and MuSV/MuLV-transformed cells could grow efficiently in agar.
Antiretroviral effect of red
microalgal polysaccharides
The polysaccharide extracted from Porphyridium
sp. was more effective in inhibiting retrovirus replication and cell
transformation by MuSV than the polysaccharides obtained from P.
aerugineum or Rhodella reticulata (Table 1).
The concentration of Porphyridium sp. polysaccharide required for
50% protection against the formation of foci of transformed cells by
MuSV or for a 50% reduction in MuLV production (as evaluated in terms of
reverse transcriptase activity) was one or two orders of magnitude lower
than that needed when P. aerugineum or R. reticulata
polysaccharide was applied. We therefore focused our research on the
anti-retroviral effects of Porphyridium sp. polysaccharide.
Porphyridium sp. polysaccharide was also
superior to other polysaccharides, such as carrageenan and dextran
sulfate 500,000, in preventing the transformation of NIH/3T3 cells.
Although the anti-transforming activity of Porphyridium sp.
polysaccharide did not seem to be much higher than that of carrageenan
or dextran sulfate at a concentration of 10 μg/ml
(Fig. 2),
at the higher concentrations of Porphyridium sp. polysaccharide
was not toxic to the cells whereas the other two biopolymers were
extremely toxic to the cells (Fig. 3).
Porphyridium sp. polysaccharide had no effect on the
proliferation of NIH/3T3 cells, even up to a concentration of 500 μg/ml
(results not shown). However, at a concentration of 1,000 μg/ml,
Porphyridium sp. polysaccharide caused the cells to stop growing
three days after the beginning of the treatment (Fig. 3).
Microscopy showed that there were there were no changes
in cell morphology in the presence of Porphyridium sp.
polysaccharide, even at a concentration of 1000 μg/ml.
Effect of dosage of Porphyridium
sp. polysaccharide on cell transformation
The algal polysaccharide significantly inhibited
malignant transformation of NIH/3T3 cells by MuSV or MuSV/MuLV. As can
be seen from Table 2,
continuous treatment with = 100 μg/ml
of Porphyridium sp. polysaccharide, from the time of infection
until 14 days after infection (scoring time), fully inhibited formation
of foci by both virus stocks.
Effect of time of addition of Porphyridium
sp. polysaccharide on cell transformation
The algal polysaccharide (100 μg/ml)
was added to NIH/3T3 cells at various times before and after infection
with either MuSV-124 or MuSV/MuLV. As can be seen from Table 3,
the polysaccharide fully inhibited formation of foci in MuSV-124- and
MuSV/MuLV-infected cultures if it was added before or at the time of
infection. If the polysaccharide was added post-infection, it was less
effective. In MuSV/MuLV-infected cultures the polysaccharide was still
significantly inhibitory to the formation of foci when added 48 h after
infection. The protective effect of the polysaccharide in these cultures
was lost only if it was added 72 h after infection, whereas in
MuSV-124-infected cultures the effectiveness of the polysaccharide was
lost when addition was as early as 48 h after infection.
The obtained differences between cell cultures infected
with either of these virus stocks (MuSV/MuLV or MuSV-124) are due to the
different characteristics of these infections. In the case of
MuSV-124-infected cell cultures, the inhibitory effect of Porphyridium
sp. polysaccharide could not be explained in terms of the inhibition of
secondary viral infections, since this virus yielded a virus
nonproducing infection [30,31].
However, in MuSV/MuLV-infected cultures, part of the inhibitory effect
of the polysaccharide against cell transformation could be a result of
inhibiting secondary viral infections, since the majority of the foci
that appeared in the cultures seemed to result from multiple secondary
infections by the progeny of the primary infection.
Effect of time of addition of Porphyridium
sp. polysaccharide on release of progeny virus from MuSV/MuLV-infected
cells
The algal polysaccharide probably exerted its effect by
preventing secondary infections, which were the major source of foci
scored in these cultures under our experimental conditions. To determine
whether the effect of the polysaccharide was due to the arrest of virus
release from the primary infected cells or merely from blocking the
establishment of secondary infections, the polysaccharide was added at
various times before and after infection, and the release of progeny
virus was followed by assaying viral reverse transcriptase activity in
aliquots taken from the culture medium at different times post
infection. The results presented in Fig. 4
showed that the infection cycle is completed within 20�24 h after
inoculation, this being the time at which the appearance of the first
progeny could be detected. The polysaccharide had a significant
inhibitory effect on the release of virus progeny even when it was added
48 h after infection. Therefore, the significant prevention of formation
of malignant foci by the polysaccharide at this late time was most
likely due to its action against the subsequent secondary infection
cycle. These results are in agreement with our suggestion that in MuSV/MuLV-infected
cultures, part of the inhibitory effect of the polysaccharide against
cell transformation could be a result of inhibiting secondary viral
infections.
Effect of time of removal of Porphyridium
sp. polysaccharide on cell transformation
To determine whether it is necessary for Porphyridium
sp. polysaccharide to remain in the culture during the whole time until
scoring of foci, cells were treated with the polysaccharide 2 h before
infection and polysaccharide was removed at various times after
infection. Foci were scored 12 days after infection. As can be seen from
Table 4,
about 75 and 65% of the transforming capacity of MuSV-124 and MuSV/MuLV,
respectively, were recovered when the polysaccharide was removed at the
time of infection. When Porphyridium sp. polysaccharide was
removed 72 h post-infection, only 55% and 20% of transforming capacity
of MuSV-124 and MuSV/MuLV, respectively, were recovered.
As a consequence of the different character of the
infection by these two virus stocks, their focus-forming capacity
responded quite differently to the timing of polysaccharide addition and
removal. In the case of MuSV/MuLV infected cultures, focus formation was
significantly inhibited even when the polysaccharide was added or
removed at longer times after infection compared to MuSV-124 infected
cultures. These findings indicate that the continuous presence of
thepolysaccharide in the culture medium after infection with the virus
was essential for full prevention of malignant transformation over the
tested period (about two weeks). When the treatment with the
polysaccharide was terminated immediately post-infection (Table 4),
there was a significant recovery in the appearance of malignant
transformed cells for all tested concentrations of the polysaccharide.
This reversibility strongly suggests that the polysaccharide, partially
at least, exerted its inhibitory effect on a certain event occurring
after proviral integration. In addition, this reversibility could not be
explained only by the possibility of preventing viral reinfections by
the polysaccharide because in the case of MuSV-124 infections there are
no reifections [30,31].
The inhibitory effect does not seem to be mediated by interferon or by
an interferon-like antiviral state, since interferon has been found to
inhibit certain events occurring before proviral integration [31].
Our results do not rule out the possibility that at
least part of the inhibitory effect of the polysaccharide was due to
blocking some of the viral receptors, thus interfering with the
penetration of the virus into the cells. This possibility was supported
by our results showing that treatment of the cells with the
polysaccharide post-infection caused a significant inhibition of cell
transformation, but that this inhibition was less impressive than that
obtained when treatment with the polysaccharide was started before or at
the time of infection (Table 3).
This possibility is also in agreement with various previous studies [9,17,18,22,24,32]
that suggested that sulfated polysaccharides prevent early steps in the
viral life cycle. In addition, some of our data not presented here
showed that the inhibitory effect of Porphyridium sp.
polysaccharide on cell malignant transformation by MuSV was not a result
of a direct interaction between the polysaccharide and the virus
particles. In contrast, our previous data (29) showed a strong
interaction between Herpes simplex virus (HSV 1 and HSV-2)
particles and Porphyridium sp. polysaccharide. This contradiction
could be due to differences in viral envelope composition. Herpes
viruses envelope is positively charged, whereas retroviruses are
negatively charged. Therefore, the sulfate groups of the polysaccharide
could easily interact with positively charged viruses.
The present data show that the red microalgal
polysaccharides profoundly inhibited retroviral malignant cell
transformation and retrovirus replication. Most effective inhibitory
activity of these polysaccarides on cell transformation was obtained
when the cells were treated with polysaccharide before or at the time of
infection. These results support the possibility that at least part of
the inhibitory effect of the polysaccharide was due to blocking some of
the viral receptors, thus interfering with the penetration of the virus
into the cells. On the other hand, the reversibility of this inhibitory
activity strongly suggests that the polysaccharide exerted its
inhibitory effect also on a certain event occurring after proviral
integration. Thus, it appears that Porphyridium sp. polysaccharide has a
pleiotropic mode of action during the infection cycle of MuSV. The exact
steps (or step) during the viral replication cycle that are affected by
Porphyridium sp. polysaccharide remain to be elucidated.
NIH/3T3 cells (mouse fibroblast cells) were grown at 37�C
in RPMI medium supplemented with 10% new born calf serum (NBCS) and the
antibiotics penicillin, streptomycin and neomycin. Clone 124 of TB cells
chronically releasing Moloney murine sarcoma virus (MuSV-124) (31) was
used to prepare a virus stock that contained an approximately 30-fold
excess of MuSV particles over Moloney murine leukemia virus (MuLV)
particles. MuLV and MuSV used in this research were grown on NIH/3T3
cells. The virus concentration was determined by counting the number of
foci (ffu-focus-forming units) in the case of MuSV and by the reverse
transcriptase assay in the case of MuLV.
Preparation and purification of
microalgal polysaccharide
Polysaccharides produced from three species of red
microalga; Porphyridium sp., P. aerugineum and Rhodella
reticulata, were used in this study. The polysaccharides were
collected and purified as previously described [33].
Briefly, these polysaccharides are produced and secreted into the growth
medium by the appropriate red microalgae. The medium was collected,
cells were removed by centrifugation and the supernatant containing the
polysaccharides was dialyzed and lyophilized.
Cell infection and determination of
viral infection
A monolayer of NIH/3T3 cells was grown in 9-cm2
tissue culture plates and treated with 0.8 μg/ml
of polybrene (a cationic polymer required for neutralizing the negative
charge of the cell membrane) for 24 h before infection with the virus.
Free polybrene was then removed, and the cells were incubated at 37�C
for 2 h with the infecting virus (MuSV-124) at various concentrations in
RPMI medium containing 2% of NBCS. The unabsorbed virus particles were
removed, fresh medium containing 2% NBCS was added, and the monolayers
were incubated at 37�C. After 2�3 days, the cell cultures were
examined for the appearance of malignant transformed cells. The amount
of malignant transformed cells was expressed as the percentage of
transformed cells in the inspection field or as the number of foci in
the infected culture 10 days after infection.
Reverse transcriptase assay
Viral reverse transcriptase activity was assayed as
previously described [34].
Acknowledgements
This research was supported by Ma'OF (established by
the Kahanoff Foundation).
We thank Ms Marion Milner for typing. and editorial
review of the manuscript.
Keith
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Red Marine Algae's Medicinal and Therapeutic Usefulness:
Over the last 25 years there has been an increased scientific
understanding of biological specificity and its subsequent relation to
the body's immune system. Current research on Dumontiaceae suggests a
breakthrough in the discovery of natural immunomodulatory and antiviral
agents.
It all started when intensive studies of marine organisms began in the
1970s to locate potential sources of pharmacologically active agents. In
a search for anti-herpetic substances, studies of California red marine
algae proved to be particularly interesting (Ehresmann et al., 1977,
1979, Hatch et al., 1979 and Richards et al., 1978). One study,
conducted by Senior Research Fellow of the chemistry department at G. D.
Searle & Co., Dr. Raphael Pappo, Ph. D., demonstrated the algae's
beneficial effects on people with Herpes Simplex Virus I and II. Several
years of study suggested to Dr. Pappo that the red marine algae assists
the body's specific immune regulatory response and plays a key role in
preventing the recurrence of the virus .
More recent research on extracts of red marine algae suggest that
specific carbohydrates (sulfated polysaccharides) may inhibit both the
DNA and RNA of viral infections and may operate both outside and within
our infected cells (Baba et al., 1988, Mitsuya et al., 1988, Ueno and
Kuno, 1987.) Work done in this area has shown that sulfated
polysaccharide compounds suppressed retroviral replication and inhibited
viral reverse transcriptases (Solomon et al., 1966, Schaffrath et al.,
1976). A study done by Neushul (1990) showed that nearly all of the 39
species of marine red algae, including the family Halymeniaceae, also
contained and exhibited an inhibitory substance that suppressed
retroviral replication and inhibited viral reverse transcriptases.
Studies by Nakashima et al., (1987, 1988) support the hypothesis that a
common immunomodulatory cell wall carbohydrate, like carrageenan, is a
type of heparin receptor molecule, binding to a cell and triggering a
specific cellular response sequence. Carrageenan may also be
internalized into infected cells, thus inhibiting the virus. It also may
inhibit fusion between infected cells Neushul (1990), Gonzales et al.,
(1987) suggesting that sulfated polysaccharides inhibit a step in viral
replication subsequent to viral internalization but prior to the onset
of late viral protein synthesis. In conclusion, the research indicates
that the polysaccharides act as an immunomodulatory agent.
Because of the severity of the present AIDS epidemic and the
debilitating effects of Herpes Simplex and Epstein-Barr, it is becoming
more important than ever to re-examine the antiviral and
immunomodulatory effects of red marine algae.
Long term relief for Herpes? Alternative treatment may help!
Historically, there has been no long term relief for chronic sufferers
of herpes simplex infections, let alone a cure. Herpes sufferers are
seemingly at the mercy of this viral menace. Despite failure at the
eradication of the herpes virus, success in the short term by
temporarily suppressing its proliferation has yielded positive results.
One such agent, acyclovir, a nucleoside analogue,has been regarded as
the drug of choice by the medical community. However, as with most
drugs, there are side effects. Are there no alternatives?
There are as many known factors which contribute to a chronic case of
herpes, while other factors remain a mystery. Finding ways to stop or
curb some of the known factors which predispose one to herpes activity
can be helpful. Chronic herpes sufferers are well accustomed to the
recommended restrictions in diet and lifestyle. Yet, even healthy
individuals who seemingly do everything right to lead a herpes-free life
cannot escape this relentless virus. So, what's next?
Treatment with acyclovir relieves symptoms, reduces the amount of
infectious virus released from the sores and speeds healing. The
treatment does not prevent subsequent attacks or diminish their
frequency or severity. The effect of acyclovir in a herpes virus
infection is to inhibit the synthesis of viral DNA. Prophylactic courses
of oral acyclovir can have a modest impact on recurrent infections, but
the cost of the drug and its potential toxicity over the long term do
not justify such regimens in most cases. In the majority of cases for
genital herpes, general recurrence patterns returned within 8 to 25 days
after stopping long term use.
Laboratory studies suggest prolonged administration of acyclovir as a
prophylactic or its prescription for trivial infections might favor the
appearance of virus strains that are both drug-resistant and pathogenic.
This concern over the advent of drug resistant pathogens, has recently
come to pass. The NIB reported that a new strain of genital herpes (HSV-II)
has evolved upon which acyclovir had no effect.
Given the drug like nature of acyclovir, with side effects included,
herpes sufferers have sought a natural approach to prevent or suppress
their herpes symptoms. The most popular natural remedy, sold in health
food stores, are high doses of the amino acid L-lysine. High doses of
L-lysine, which is an essential amino acid, have been clinically shown
to suppress the proliferation of the herpes virus. Earlier research
revealed that some amino acids increased growth in viral activity and
others decreased such activity. Further studies showed that one could
effectively alter the chemistry of the cellular environment by
increasing the availability of a particular amino acid. In the case of
L-lysine, inducing a higher concentration of L-lysine was shown to lower
the arginine cellular concentration. The effect of depleting the
existing reserves of arginine (a non-essential amino acid) combined with
the presence of L-lysine effectively thwarts assembly of viralprotein
coats. Without this vital structural component, herpes viruses cannot
invade new cells. Potential herpes infections are thus temporarily
aborted.
Acyclovir and L-lysine, although widely used, have provided variable
success for its users. The fact that known side effects from taking
acyclovir include nausea, vomiting, diarrhea, dizziness and headache are
not encouraging given that effective treatment of acyclovir requires
daily use. Also, little is known about the long term effects and
toxicity. One study showed chromosome damage when taking large doses
even though low dosages are considered safe. L-lysine, once announced as
a major medical breakthrough in the prevention of herpes disease, has
its downside as well. Research has shown that a decrease in arginine
lowers lymphocyte immune reactivity in healthy human beings.
Essentially, an increase in daily intake of L-lysine has the net effect
of lowering our natural immunity due to the decrease of arginine in the
cellular environment (perhaps arginine, once thought non-essential is
becoming increasingly essential for our own survival). The fact that it
suppresses herpes simplex viral activity is significant, but not at the
expense of our adaptive immune system. Neither acyclovir nor L-lysine
are recommended for long term prophylactic treatment. Individuals
seeking a dailymaintenance dosage to ward off herpes outbreaks would be
ill advised to relyon L-lysine or acyclovir. Chronic herpes sufferers
would be better off to investigate other means to prevent or suppress
their herpes condition. Is there no hope?
Western medicine, armed with its infinite technological powers, can
still help us. Many potent botanical agents have been investigated but
never made it through the arduous process of drug approval. Difficulties
in understanding the intricate process under which particular botanical
agents interact within the human body has kept many useful medicines
from ever reaching the people who most urgently need them. In addition,
many botanical agents can only work in their whole plant form. They work
on multiple levels and act synergistically within the body.
Although the actions of these botanical agents in whole plants (commonly
described as herbs or medicinal plants) are difficult to trace and
report scientifically, a close monitoring of clinical results by trained
practitioners can be useful and show efficacy. Certainly, using our
powers of observation to determine whether a particular treatment works
better than no treatment, or better than some other treatment for a
patient whose health status and history is well documented can be
significant.
One such casualty of the drug approval process is red marine algae.
Research on antiviral carbohydrates from marine red algae indicate a
high potential for low-cost, broad spectrum antiviral agents. Further
research into Red Marine Algae produced two patents where clinical
efficacy for herpes I and II was clearly shown. The treatment was
effective for treating subjects (e.g. human patients) both prior to and
subsequent to herpes infection. It was used topically to alleviate
symptoms associated with herpes infections or preferably systemic, by
oral administration, to eradicate the virus and thereby prevent symptom
recurrence. No side effects or toxicity were noted. This treatment,
which now must be considered alternative, suggests a breakthrough in the
discovery of natural immunomodulatory and antiviral agents.
Recent research and gathering of anecdotal evidence on the health
benefits and antiherpetic action of red marine algae has yielded much
promise. Its use as a topical has been further documented and thought
superior to acyclovir. It was shown to be clinically effective against
herpes zoster infections as well. Anecdotal reports from patients
suffering from Epstein Barr (another herpes virus) and Candida have
shown marked improvement in a short period of time through oral
administration (systemic).
General health benefits show red marine algae useful in weight-loss
programs and for lowering cholesterol and fat in the blood. It contains
soothing, mucilaginous gels such as algin, carregeenan, and agar, which
specifically rejuvenate the lungs and gastrointestinal tract. Once
thought of as a liability that blocked assimilation, the tough cell wall
in Dumontiaceae has been found to be invaluable. It binds with heavy
metal, pesticides, and carcinogens, and carries these toxins safely out
of the body. Contained within the cell walls are simple sugars called
complex polysaccharides. These long chained complex sugars stimulate
interferon production as well as other anti-tumor and immune-enhancing
activity (improving activity of T- and B-cells). Other compounds in the
cell wall are related to those found in friendly bacteria which fortify
and strengthen our immune systems to fight against invading organisms
and toxins.
Although the effects of long term use of an alternative treatment such
as the red marine algae, Dumontiaceae, has not been clinically
substantiated, edible seaweeds have been consumed for thousands of years
and are considered safe, nutritious, and beneficial. The added dimension
that science has uncovered surrounding its antiviral and
immunomodulatory potential; opens up a whole new source of food that
could serve to palliate or even hopefully cure virally caused diseases.
Since most life derived from the sea, the novel idea that the ocean lies
untapped as perhaps our greatest medicinal resource is entirely possible
and may be critical to our human survival.
Therapeutic Application for Newly Discovered Marine Algae
Researchers in the mid seventies and early eighties were exploring rare
algae that potentially modeled immunomodulatory activity in humans.
Investigations revealed some thirty species which enhanced the immune
systems's regulatory response and were shown to be antiviral. The more
promising part of this discovery was the antiviral specificity of each
species towards a variety of pathogens.
Current research on a red marine algae has exhibited promising results
in controlling and reducing both Candida and Herpes Simplex Virus
populations. Patients have reported a stopping or lessening of growth
within the body. Researchers believe these special algae may serve as a
gateway to resist or even cure many bacteria, fungi, or and viral
pathogens.
Could algae, commonly known as ocean vegetables, be one of the most
important new therapeutic food? Scientific research has only reinforced
the medicinal and nutritional importance of ocean vegetables. Numerous
cultures have used ocean vegetables to complement their healthy diet.
Ocean vegetables were most commonly used to prevent aging and prolong
life. Since all life evolved from the sea, we may think of the ocean as
a vast nutritional soup that lies untapped as perhaps our greatest
medicinal resource.
Conclusion
The powers of ocean vegetables has been sought for thousands of years
for their ability to prolong life, prevent disease, and enhance life.
Ocean vegetables contain ten to twenty times the minerals of land
plants, as well as an abundance of vitamins and other elements necessary
for proper metabolism. Each ocean vegetable exhibits a distinct nutrient
profile and a selective nature for its medicinal use. Current research
has now established a link between nutrient-rich red marine algae and
the body's immune system response.
Our ability to survive in a hostile environment that may seem out of
control demands that we take steps to recover our health and maintain
our immunity. Therein ocean vegetables may be one of our most important
allies in a changing world.
References:
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3. Cole and Sheath, (Ed.), Biology of the Red Algae, Cambridge
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These statements have not been evaluated by the Food and Drug
Administration. The products found here are are not intended
to diagnose, treat, cure, or prevent any disease. This data is for
information only.
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Last modified: August 27, 2002
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