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Medical Forum / Diseases and Disorders / Herpes / November 2005

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L-Canavanine

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Perl Molson - 31 Oct 2005 15:21 GMT
What is it?

L-Canavanine is one of over 300 documented non-protein amino acids,
also known as secondary metabolites or anti-metabolites, which exist in
plants (most commonly in legumes and their seeds). Essentially, they
assist in the breakdown of the 20 known amino acids common in both
plants and animals. L-Canavanine's chemical structure is quite
complex and defined as follows:

H2N-C(=NH)-NH-O-CH2-CH2-CH2-CH(NH2)CO2H

Specifically, L-Canavanine is an L-arginine antimetabolite. In the
Sutherlandia plant, L-Canavanine is present at levels of just over 2.0
mg per dry leaf gram. It acts as an anti-predatory agent in the plant,
protecting it against various insects.

L-Canavanine has been shown to be an inhibitor (selective) of inducible
nitric oxide synthase (iNOS), with possible usage in the treatment of
alcohol induced brain damage and certain other neurological disorders.

What are its benefits?

L-Canavanine has been identified by G. A Rosenthal, of the Biochemical
Ecology Laboratory at The University of Kentucky, as a likely
therapeutic agent for human pancreatic and some other cancer types. It
has been demonstrated by N. Suzuki, A. Sakomoto and R. Ogawa as having
strong potential for the treatment of septic shock and chronic
inflammation. These are just a few of numerous research papers on the
substance.

There are a number of patents associated with L-Canavanine which
indicate it's applicability in the treatment in a number of ailments.
Two of the main examples are: Crooks, Rosenthal and Gerald were granted
a patent in 1994 for the use of the substance to treat pancreatic
cancer (Patent No. 5,552,440) and Karson, Lyon and Freeman being
granted a patent that uses the substance in the treatment of numerous
brain disorders in 1996 (Patent No. 5,527,825). In patent No.
6,346,519, E. Petrus gives L-Canavanine a partial role in the treatment
of arthritis.

More research is currently underway on the substance and its
applicability to certain other conditions. Details will be placed here
as soon as confirmed.

Is a patent total proof that L-Canavanine is effective?

The tests, experiments and theory associated with the above have met a
defined standard of proof with the US Patent office, as evidenced by
the awarding of the patents. Despite this, we do not regard the
awarding of the patents as demonstrable evidence for the efficacy of
L-Canavanine in themselves. But critically, the above findings do
correlate with the widespread anecdotal evidence we have taken from
individuals who have used Sutherlandia and the fact that their
ancestors, going back hundreds of years, have also used the plants for
the same purpose, with very positive results.

Are there any side effects?

If taken in severe overdose quantities (i.e. 1,000 mg per day for 12
months), L-Canavanine can cause thinning of the blood. Furthermore, it
can exacerbate the symptoms of Lupus, a disease where the body's own
immune system starts to attack various organs. Individuals with kidney
conditions must consult their physician before taking this substance.

At recommended dosage levels, the L-Canavanine in Sutherlandia
Frutescens Tablets has not been shown to cause side effects.
Perl Molson - 31 Oct 2005 15:41 GMT
Effects of canavanine treatment on herpesvirus morphogenesis in
cultured cells.

Smith JD, Moore DM.

L-Canavanine, a naturally occurring analog of arginine, effectively
inhibited the morphogenesis of herpes simplex virus (HSV) and human
cytomegalovirus (HCMV) when added at the time of infection, but allowed
the expression of several cytopathic changes. Exposure to canavanine at
progressively later times ultimately led to qualitatively normal virion
maturation. Under no conditions were morphologically aberrant viral
particles observed. However, HSV-infected cells treated at 3 h
postinfection or later did contain distinctive cytoplasmic inclusions
resembling HCMV dense bodies. Recovery experiments showed that
HCMV-infected cells exposed to canavanine for 1-5 days could support
normal viral morphogenesis when washed free of this agent.

PMID: 6282774 [PubMed - indexed for MEDLINE]

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstra
ct&list_uids=6282774&query_hl=2

Perl Molson - 31 Oct 2005 16:06 GMT
Specificity in protein biosynthesis

The general problem of recognition of their substrates by the
aminoacyl-tRNA synthetases has been the subject of intensive research
for several decades and, with the advent of molecular biological
techniques together with available X-ray crystallographic structures,
considerable progress towards the understanding of tRNA recognition has
been achieved. However, investigations into the recognition of the
amino acid and the mechanisms involved in guaranteeing accuracy during
aminoacylation have, with the tools of recombinant DNA, only recently
been reinitiated, following detailed biochemical studies in the
mid-'70s. These defined the pathways of "chemical proofreading" or
"hydrolytic editing" by which a transiently activated "wrong" amino
acid is corrected by the enzyme itself, in a proofreading step.

Renewed interest in recent years, with the possibilities available
through the methods of genetics and molecular biology, boosted the hope
that attempts at solving this central enzymological puzzle may be able
to pinpoint regions or even residues which are essential for
maintaining fidelity. Mutational analysis established conserved
residues that are important for proofreading. However, it is quite
possible, based on phenomenological observations, that several
different mechanisms may have evolved.

It is reasonable then, to approach the elucidation of this/these
mechanism/s from an alternative angle; using the systems provided by
Nature, in the form of inhibitory non-protein amino acids. A comparison
between the enzyme from a non-producer, from a producer and from a
resistant seed predator (using the possibilities offered by recombinant
techniques to obtain well-defined, purified components) will give
important information on how nature has solved the problem of not
incorporating these non-protein amino acids into the growing protein
chain. Of the thousands of known non-protein amino acids (NPA), about
300 occur in plants. They are found mostly in a small number of
families, such as the Leguminosae, Cucurbitaceae, Sapindacae, Aceraceae
and Hippocastenaceae. Many of these NPA are structurally similar to the
components of common proteins. The incorporation of NPAs into proteins
may be associated with autoimmune diseases in humans. Furthermore,
there is evidence of a phenotypic conversion of ras-transformed human
cells to normal due to incorporation of the tyrosine analogue,
azatyrosine, into cellular protein. One NPA that has received some
attention is canavanine, (L-2-amino-4-(guanidinooxy)butyric acid), the
guanidinooxy structural analogue of arginine (Fig. 1).

Fig. 1: L-Arginine (top), L-Canavanine (bottom).

Since its discovery in the 1920's in the seeds of jack bean (Canavalia
ensiformis), massive accumulation of this toxic secondary metabolite
has been established in the seeds of at least 1500 members of the
Lotoideae; a major subdivision of the Fabaceae. Canavanine storage in
the seeds of such plants can account for more than 10% of the seed dry
matter. The preponderance of seed canavanine, its depletion during
growth of the seedlings, its extreme toxicity to actual (insects,
rodents) and serendipitous (humans) seed predators, and its high
nitrogen content all point to the conclusion that the plant has
constructed a compound that is simultaneously a storage compound and a
major seed defence. This strategy is commonplace in legume seeds with a
variety of secondary compounds. The evolutionary history is undoubtedly
that of evolution of toxicity in compounds that were initially purely
storage in function; a change with obvious adaptive value. The toxicity
of canavanine to a non-producer species ranging from bacteria to
insects, to plants and mammalian cells has been extensively described.
Canavanine binds to nearly every enzyme that utilizes arginine. The
mechanism of its toxicity has, however, been localized to its
incorporation into proteins giving rise to functionally aberrant
polypeptides. The loss or reduction in activity in
canavanine-containing proteins has been observed for antibacterial
proteins and for lysozyme into which canavanine had been randomly
incorporated. Canavanine containing proteins are also preferentially
degraded, in vivo. Allende and Allende in the '60s first documented the
attachment of canavanine to tRNAArg by the rat liver arginyl-tRNA
synthetase.

This lack of discrimination by an enzyme belonging to a family of
enzymes whose accuracy in recognizing their substrate is essential in
maintaining an effective protein synthesizing machinery, was confirmed
for the enzyme from E.coli and Neurospora while an assay with a crude
extract from the producer, Canavalia, showed a much reduced activity.

The accumulation of canavanine in producers provides nitrogen storage
for the growing seedling and also provides an effective protective
chemical barrier to predation and disease and raises the question as to
how the producer itself avoids autotoxicity. In addition to the need
for a detoxifying mechanism for the producer being self-evident, there
are also organisms that are themselves non-producers but have
specialized in feeding on the producer. Such a situation has been found
in the bruchid beetle Caryedes brasiliensis whose larvae infest and
feed on Dioclea violacea seeds containing up to 13% canavanine.

In this project, we propose to isolate, clone and overexpress the
gene(s) for arginyl-tRNA synthetase from at least two canavanine
producers (Canavalia ensiformis, Dioclea violaceae (formerly,
megacarpa)), a closely related non producer (Glycin max) and a
resistant seed predator (Caryedes brasiliensis). We will study the
structure/function of the gene products in order to identify the
molecular mechanism(s) involved in this example of allelochemical
detoxification while establishing the discriminatory elements involved
in the recognition of arginine/canavanine by arginyl-tRNA synthetase.
Additionally, should the adult Caryedes prove to be canavanine
sensitive, by studying the argS genes/regulatory elements/gene products
we intend to obtain information on the developmental regulation that
occurs during insect metamorphosis generating a canavanine-resistant
(Caryedes larvae) or non-resistant (Caryedes adult) arginyl-tRNA
synthetase.

We have used PCR and RACE methods to isolate and characterise the
full-length cDNA sequence from Canavalia. The genomic sequence is
currently being established. We have compared the derived protein
sequence with over 50 sequences in the databases (all non-producers,
including Arabidopsis and Rice) and with the crystal structure of the
enzyme/arginine complex from yeast. Our results indicate, that the
Canavalia sequence is phylogenetically considerably removed from the
yeast sequence, so that a modelling on the basis of the yeast
structure, while technically possible using the Spdb, need not reflect
the true situation in detail. However, the amino acids defined in yeast
as being responsible for arginine binding are conserved in Canavalia
(as in all other species).

We have been fortunate in establishing a collaboration with the
tropical ecologist Dr. D. Janzen (University of Pennsylvania) who is
uniquely able to provide specimens of the neotropical bruchid beetle
Caryedes brasiliensis and its host plant (Dioclea violacea) from his
field studies in Costa Rica, as well as decades of experience with
these and other canavanine resistant organisms. He is committed to
providing specimens and natural history information from the field, in
Costa Rica.

Updated: November 2004
http://www.biologie.uni-freiburg.de/data/bio3/igloi/Accuracy.htm
Perl Molson - 31 Oct 2005 16:19 GMT
Later, research of our chair demonstrated that LO exhibited antiviral
activity. It inhibited the reproduction of type I Herpes simplex virus.
In the presence of 10-2 U/ml (0.3 µg/ml) of the enzyme, the cytopathic
effect of type I Herpes simplex virus on the cells cultivated in vitro
decreased almost 105-fold. Besides, LO effectively suppresses the
expression of the viral antigens. Therefore, it was concluded that
highly purified preparations of LO are more effective agents
suppressing the reproduction of Herpes simplex virus compared to
luteolin and acyclovir [36, 37].

Abbreviations: LO) L-lysine alpha-oxidase.

http://www.protein.bio.msu.ru/biokhimiya/contents/v67/full/67101394.html
Perl Molson - 01 Nov 2005 15:42 GMT
Now that's an interesting amino-acid, L-Canavanine (found in alfalfa
sprouts, see below).
Herpes may take it as L-Arginine.
Apparently, Astragalus Root contains L-Canavanine, as well (hmmm, maybe

that's why it works so well against herpes)

http://www.peacefulmind.com/safety.htm
ALFALFA (Medicago sativa)

Safety: Alfalfa in its various forms may present some health risks.
Powdered alfalfa herb, alfalfa sprouts, and alfalfa seeds all contain
L-cavanine, a substance that may cause abnormal blood cell counts,
spleen enlargement, or recurrence of lupus in patients with controlled
disease. However, heating alfalfa may correct this problem.
Researchers investigating alfalfa seeds' ability to lower cholesterol
levels discovered that it had another effect on the lab animals used
for testing. In some of the monkeys, it caused a disease very similar
to lupus. Further research on this effect revealed that monkeys that
had abnormal blood cell counts when eating either alfalfa seeds or
sprouts, and then recovered when alfalfa was no longer part of their
diet, developed the symptoms again when given an isolated component of
alfalfa called L-canavanine. Alfalfa seeds and sprouts have a higher
concentration of L-canavanine than the leaves or roots.
In a clinical trial of alfalfa seeds for lowering cholesterol involving
only three human volunteers, one man who participated developed
pancytopenia (an abnormally low number of all of the various types of
blood cells) and enlargement of the spleen. Additionally, there are two
published case reports of patients who had lupus which was controlled
with drug therapy, suffering relapses after consuming alfalfa tablets.
Again, L-canavanine is thought responsible for these effects.
When alfalfa seeds were autoclaved (heated to extremely high
temperatures) and fed to monkeys for a year, no ill effects were seen,
and the monkeys' cholesterol levels decreased. It may be that the
L-canavanine can be destroyed by extreme heat, while the saponins that
seem to be responsible for the beneficial effects of alfalfa remain
intact. If so a heat-treated product might prove safe; however, much
research remains to be done before we can know this for certain.
At present, it seems prudent that people who have been diagnosed with
lupus, or those who suspect a predisposition to it based on family
history, should probably avoid alfalfa. This includes the tablets used
for supplements and the sprouts on the salad bar (go for the lettuce or
the spinach instead).
Because of the estrogenic effects of some of alfalfa's components,
alfalfa is not recommended for pregnant or nursing women or young
children. In addition, the high vitamin K content in alfalfa could, in
theory, make the drug warfarin (Coumadin) less effective.
Finally, a number of cases of food poisoning have been documented from
fresh sprouts infected with bacteria that was present on the seeds
prior to germination. Unfortunately, sprouts can appear fresh and yet
host enough bacteria to cause illness in people who eat them. Some
health care workers recommend that those at higher risk for such
infections-young children, those with chronic diseases, and the
elderly-avoid eating sprouts altogether.

Dosage:A typical dose of alfalfa for tea is 1 to 2 teaspoons per cup,
steeped in boiling water for 10 to 20 minutes. Tablets and capsules of
whole alfalfa or alfalfa extracts should be taken according to the
manufacturer's recommendations. Certain products are said to be free of
canavanine and other potentially harmful constituents, and may be
preferable.

Contraindications: If you are taking warfarin (Coumadin), the high
vitamin K content of alfalfa might make it less effective.

NATURAL TOXINS IN BEANS ARE NO CAUSE FOR ALARM
by Steve Meyerowitz
People have been alarmed by articles alleging that alfalfa and other
sprouts contain toxins. This news has struck fear in Vegetarians and
others because sprouts have always been considered "health food." Most
of us have heard this news from the very popular author and proponent
of natural healing, Andrew Weil. When questioned for more detail, Dr.
Weil refers us to well known toxicologist Bruce Ames. Ames has written
extensively on plant toxins. In a recent telephone interview, Dr. Ames
claimed that he is not against sprouts. "Eat any sprouts you want..."
he said. "There's nothing wrong with sprouts....If I was to be against
sprouts then I would be against every kind of vegetation."

Ames tells us that there are toxins in the seeds of many common plants
and vegetables as a natural protective mechanism against insects and
other predators. The natural toxin identified in sprouting seeds is
known as L-canavanine, a pre-cursor necessary for the development of
the amino acid arginine. L-canavanine is one of 600 non-protein amino
acids synthesized by plants. It is found in hundreds of legumes (beans)
and other arginine rich foods such as garlic and onion.

However, it's presence in beans (alfalfa is a tiny bean) is not a cause
for alarm. During the germination process, L-Canavanine is converted to
Arginine, one of the essential amino acids. In 1960, Dr. E.A. Bell,
demonstrated that once alfalfa reaches the green stage, this toxin is
reduced to a trace amount. More recently, L-Canavanine biochemist
Gerald A. Rosenthal of the University of Kentucky found that this
natural toxin "was shown to exhibit significant antineoplastic activity
against MIAPaCa-2, a human pancreatic cancer." This natural plant
protectant can also protect us against cancer! This work is underscored
by that of John Hopkins researcher Paul Talalay who in 1997 found that
the cancer protectant sulphoraphane is present in broccoli sprouts in
quantities up to 50 times greater than in broccoli.

The research quoted on this subject involved feeding monkeys
L-canavanine sulfate tablets plus biscuits made from raw, unsprouted
alfalfa beans and "sprouts." However, none of this research actually
involved alfalfa sprouts as they are typically consumed by people. The
"sprouts" were beans germinated for 1-3 days and then oven dried. They
never turned green. Sprouts for human consumption are grown for
approximately 7 days and contain only a trace of L-canavanine.

The thrust of the research was to explore the connection between this
toxin and the auto-immune disease Lupus. It was not a test of alfalfa
sprouts. In fact, "sprouts" played only a marginal role in 2 of the
numerous studies on this toxin between done between 1981 and 1984.
According to Lupus specialist Dr. Emil J. Bardana, who worked with Dr.
Rene Malinow on the two studies involving the so called "sprouts,"
there is no basis to say that eating alfalfa sprouts would cause lupus
or SLE. "I wouldn't discourage my lupus patients from eating alfalfa
sprouts." The amount of sprouts you eat on a salad "isn't anywhere near
the dose we fed the monkeys."

Keeping things in perspective, many common foods contain toxins all of
which are consumed in greater quantities than sprouts: potatoes have
solanine, mushrooms have strychnine, apples and apricots have cyanide.
If you believe such toxins threaten our health, then you should also
eliminate tomatoes, eggplants and peppers, mustard, cottonseed oil,
peanuts, black pepper, root beer (sassafras) and last but not least,
coffee which, according to Dr. Ames, contains the "natural mutagen
chlorogenic acid and highly toxic atractylosides".

Let us not forget that as living plants, sprouts are some of our
richest sources of phytochemicals, enzymes, anti-oxidants,
nitrosamines, trace minerals and chemo-protectants such as
sulphoraphane and isoflavone which work against toxins, resist cell
mutation and invigorate the body's immune system. Don't doubt the
sprout!

"Until Man duplicates a blade of grass, nature can laugh at his so
called scientific knowledge." --Thomas Edison

©1998 Steve Meyerowitz

-BACK-

http://www.sproutman.com/toxins.html

> Later, research of our chair demonstrated that LO exhibited antiviral
> activity. It inhibited the reproduction of type I Herpes simplex virus.
[quoted text clipped - 9 lines]
>
> http://www.protein.bio.msu.ru/biokhimiya/contents/v67/full/67101394.html
Perl Molson - 02 Nov 2005 17:03 GMT
Some additional readings:

http://www.pubmedcentral.nih.gov/pagerender.fcgi?artid=1066589&pageindex=1#page

l-Arginine and l-Canavanine Metabolism in Jack Bean, Canavalia
ensiformis (L.) DC. and Soybean, Glycine max (L.) Merr. 1

http://www.protein.bio.msu.ru/biokhimiya/contents/v67/pdf/bcm_1152.pdf
LLysine aOxidase: Physicochemical and Biological Properties
> Now that's an interesting amino-acid, L-Canavanine (found in alfalfa
> sprouts, see below).
[quoted text clipped - 152 lines]
> >
> > http://www.protein.bio.msu.ru/biokhimiya/contents/v67/full/67101394.html
 
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