http://groups.yahoo.com/group/aspartameNM/message/937
aspartame, methanol, formaldehyde, formate toxicity on rat cells:
Oyama, Akaike, et al, Jan 2002: Gold: Murray 1.3.3 rmforall:
FDA Docket 02P-0317 Recall Aspartame as a Neurotoxic Drug

Please post this to the FDA Dockets website.

Rich Murray, MA Room For All rmforall@...
1943 Otowi Road, Santa Fe, New Mexico 87505 USA 505-986-9103

Cell Biol Toxicol 2002; 18(1): 43-50
Cytotoxic effects of methanol, formaldehyde, and formate on
dissociated rat thymocytes: a possibility of aspartame toxicity.
Oyama Y, Sakai H, Arata T, Okano Y, Akaike N, Sakai K, Noda K.
Laboratory of Cellular Signaling, Faculty of Integrated Arts and
Sciences, The University of Tokushima, Japan.
oyama@...

[ Oyama Y and Akaike N have cooperated in competent cellular toxicity
studies since 1981. This is their only report on these toxicants.
It is risky indeed to hazard audacious extrapolations from the damage
of one-time exposure of cells directly to toxicants to the vastly more
complex toxic and immunological processes in whole bodies of diverse,
individual humans subject to lifelong exposure to simultaneous
barrages of chemicals. An industry PR outfit would pay a lot for a
scientific sound bite like:
"It is suggested that aspartame at abuse doses is harmless to humans."
Note that they admit the key fact that aspartame means dealing with
the toxicity of methanol, formaldehyde, and formate. Rats are about ten
times more resistant to the toxicity of methanol and its metabolites
than are monkeys and humans. ]

Aspartame is a widely used artificial sweetener added to many soft
beverages and its usage is increasing in health-conscious societies.
Upon ingestion, this artificial sweetener produces methanol as a
metabolite.

In order to examine the possibility of aspartame toxicity, the effects
of methanol and its metabolites (formaldehyde and formate) on
dissociated rat thymocytes were studied by flow cytometry.

While methanol and formate did not affect cell viability in the
physiological pH range, formaldehyde at 1-3 mmol/L started to induce
cell death. Further increase in formaldehyde concentration produced a
dose-dependent decrease in cell viability.

Formaldehyde at 1 mmol/L or more greatly reduced cellular content of
glutathione, possibly increasing cell vulnerability to oxidative
stress.

Furthermore, formaldehyde at 3 mmol/L or more significantly increased
intracellular concentration of Ca2+ ([Ca2+]i) in a dose-dependent
manner.

Threshold concentrations of formaldehyde, a metabolite of methanol,
that affected the [Ca2+]i and cellular glutathione content were
slightly higher than the blood concentrations of methanol previously
reported in subjects administered abuse doses of aspartame.

It is suggested that aspartame at abuse doses is harmless to humans.
PMID: 11991085 [PubMed - in process]
**********************************************************************

Oyama Y, Chikahisa L, Ueha T, Kanemaru K, Noda K.
Ginkgo biloba extract protects brain neurons against oxidative stress
induced by hydrogen peroxide.
Brain Res. 1996 Mar 18; 712(2): 349-52.
PMID: 8814913 [PubMed - indexed for MEDLINE]

Arata T, Oyama Y, Tabaru K, Satoh M, Hayashi H, Ishida S, Okano Y.
Cytotoxic effects of triphenylbismuth on rat thymocytes: comparisons
with bismuth chloride and triphenyltin chloride.
Environ Toxicol. 2002 Oct; 17(5): 472-7.
PMID: 12242678 [PubMed - in process]

Arata T, Oyama Y, Tabaru K, Satoh M, Hayashi H, Ishida S, Okano Y.
Cytotoxic effects of triphenylbismuth on rat thymocytes: comparisons
with bismuth chloride and triphenyltin chloride.
Environ Toxicol. 2002 Oct; 17(5): 472-7.
PMID: 12242678 [PubMed - in process]

Oyama Y, Chikahisa L, Kanemaru K, Nakata M, Noguchi S, Nagano T,
Okazaki E, Hirata A.
Cytotoxic actions of FTY720, a novel immunosuppressant, on thymocytes
and brain neurons dissociated from the rat.
Jpn J Pharmacol. 1998 Apr; 76(4): 377-85.
PMID: 9623716 [PubMed - indexed for MEDLINE]

Okazaki E, Chikahisa L, Kanemaru K, Oyama Y.
Flow cytometric analysis of the H2O2-induced increase in intracellular
Ca2+ concentration of rat thymocytes.
Jpn J Pharmacol. 1996 Aug; 71(4): 273-80.
PMID: 8886924 [PubMed - indexed for MEDLINE]

Oyama Y, Carpenter DO, Ueno S, Hayashi H, Tomiyoshi F.
Methylmercury induces Ca(2+)-dependent hyperpolarization of mouse
thymocytes: a flow cytometric study using fluorescent dyes.
Eur J Pharmacol. 1995 Jul 1; 293(2): 101-7.
PMID: 7589223 [PubMed - indexed for MEDLINE]

Oyama Y.
Modification of voltage-dependent Na+ current by triphenyltin, an
environmental pollutant, in
isolated mammalian brain neurons.
Brain Res. 1992 Jun 26; 583(1-2): 93-9.
PMID: 1324096 [PubMed - indexed for MEDLINE]

Oyama Y, Akaike N.
Triphenyltin: a potent excitatory neurotoxicant. Its reciprocal
effects
on voltage-dependent Na and K currents of mammalian brain neuron.
Neurosci Lett. 1990 Nov 13; 119(2): 261-4.
PMID: 2280903 [PubMed - indexed for MEDLINE]

Akaike N, Nishi K, Oyama Y.
Inhibitory effects of propranolol on the calcium current of Helix
neurones.
Br J Pharmacol. 1981 Jun; 73(2): 431-4.
PMID: 6263388 [PubMed - indexed for MEDLINE]
***********************************************************************

Subject: Re: ANM: debate about formaldehyde from methanol, the 11%
component of aspartame: Murray: 12.18.2 rmforall
Date: Wed, 18 Dec 2002 18:11:56 -0500
From: <mgold@...>
To: Rich Murray <rmforall@...>

On Wed, 18 Dec 2002 02:39:43 -0700 Rich Murray <rmforall@...> wrote:

> I was not able to find the journal with the Oyama (2002) study, of
> which abstract Morrow and Sullivan generously
> quoted-- it seems to me to be a classic case in which a
> "pro-aspartame" study gives details that are hardly reassuring.

Rich,

The Oyama study applies only to dissociated rat thymocyte cells. If you
are walking down the street and you happen upon a group of dissociated
rat thymocytes exposing themselves to formaldehyde at concentrations
below 1 mmol/L, you can reassure them that they are in "no danger" as
long as the exposure is no longer than 3 hours long. :-)

The study does not address the differences in toxicity of methanol and
formaldehyde between humans and rodents. Methanol is ~10 times more
acutely toxic in humans than rodents. The researchers did not adjust for
differences in toxicity and therefore their results can easily be off by
a factor of 10 or more. See:
http://www.holisticmed.com/aspartame/abuse/methanol.html#chronic

Aside from their figures only applying to rats (and having nothing to do
with humans) the researchers compared their results to results obtained
from a 1981 study of aspartame that used a methanol measurement
technique developed in 1969 that would not be able to measure changes
less than a 4-fold increase above the baseline level. See:
http://www.holisticmed.com/aspartame/abuse/methanol.html#hiding

Of course, they also don't take into account the potential accumulation
of formaldehyde adducts (since the 1981 study they are comparing their
figures to used only a single ingestion of aspartame).

The cells in the "test tube" were exposed to the test substances
(methanol, formate, or formaldehyde) for a total of 3 hours.

Their conclusions appear to be based on a few aspartame industry human
studies. I will give them the benefit of the doubt and assume that they
didn't come to their conclusions based on a test that exposes rat cells
in a test tube to one aspartame metabolite for a short time. - Mark

http://www.HolisticMed.com/aspartame 603-225-2100
Aspartame Toxicity Information Center Mark D. Gold
mgold@... 12 East Side Drive #2-18 Concord, NH 03301
http://www.holisticmed.com/aspartame/abuse/methanol.html
"Scientific Abuse in Aspartame Research"
************************************************************************

http://groups.yahoo.com/group/aspartameNM/message/932
aspartame: methanol, formaldehyde, formic acid toxicity:
brief review: Murray 12.30.2 rmforall:

http://groups.yahoo.com/group/aspartameNM/messages
for 937 posts in a public searchable archive

It is certain that high levels of aspartame use, above 2 liters daily
for months and years, must lead to chronic formaldehyde-formic acid
toxicity, since 11% of aspartame (1,120 mg in 2L diet soda, 5.6 12-oz
cans) is 123 mg methanol (wood alcohol), immediately released into the
body after drinking (unlike the large levels of methanol locked up in
molecules inside many fruits), then quickly transformed into
formaldehyde, which in turn becomes formic acid, both of which in
time become carbon dioxide and water-- however, about 30% of the
methanol remains in the body as cumulative durable toxic metabolites of
formaldehyde and formic acid-- 37 mg daily, a gram every month.
If 10% of the methanol is retained as formaldehyde, that would give 12
mg daily formaldehyde accumulation, about 60 times more than the 0.2 mg
from 10% retention of the 2 mg EPA daily limit for formaldehyde in
drinking water.

Bear in mind that the EPA limit for formaldehyde in
drinking water is 1 ppm,
or 2 mg daily for a typical daily consumption of 2 L of water.

http://groups.yahoo.com/group/aspartameNM/message/835
RTM: ATSDR: EPA limit 1 ppm formaldehyde in drinking water July 1999
5.30.2 rmforall

J. Nutrition 1973 Oct; 103(10): 1454-1459.
Metabolism of aspartame in monkeys.
Oppermann JA, Muldoon E, Ranney RE.
Dept. of Biochemistry, Searle Laboratories,
Division of G.D. Searle and Co. Box 5110, Chicago, IL 60680
They found that about 70% of the radioactive methanol in aspartame put
into the stomachs of 3 to 7 kg monkeys was eliminated within a day as
carbon dioxide in exhaled air and as water in the urine. They did not
mention that this meant that about 30% of the methanol must transform
into formaldehyde and then into formic acid, much of which must remain
as toxic products in all parts of the body. They did not report any
studies on the distribution of radioactivity in body tissues, except
that blood plasma proteins after 4 days held 4% of the initial
methanol. This study did not monitor long-term use of aspartame.

This long-term low-level chronic toxic exposure leads to typical
patterns of increasingly severe complex symptoms, starting with
headache, fatigue, joint pain, irritability, memory loss, and leading to
vision and eye problems and even seizures. In many cases there is
addiction. Probably there are immune system disorders, with a
hypersensitivity to these toxins and other chemicals.

Confirming evidence and a general theory are given by Pall (2002):
http://groups.yahoo.com/group/aspartameNM/message/909
testable theory of MCS type diseases, vicious cycle of nitric oxide &
peroxynitrite: MSG: formaldehyde-methanol-aspartame:
Martin L. Pall: Murray: 12.9.2 rmforall

http://groups.yahoo.com/group/aspartameNM/message/915
formaldehyde toxicity: Thrasher & Kilburn: Shaham: EPA: Gold: Murray:
Wilson: CIIN: 12.12.2 rmforall

http://www.drthrasher.org/formaldehyde_1990.html [ full text ]
Arch Environ Health 1990 Jul-Aug; 45(4): 217-23
Immune activation and autoantibodies in humans
with long-term inhalation exposure to formaldehyde.
Thrasher JD, Broughton A, Madison R.
Thrasher & Associates, Northridge, California.

Arch Environ Health 2001 Jul-Aug; 56(4): 300-11
Embryo toxicity and teratogenicity of formaldehyde. [ 100 references]
Thrasher JD, Kilburn KH.
Sam-1 Trust, Alto, New Mexico, USA.
http://www.drthrasher.org/formaldehyde_embryo_toxicity.html
[ 127K full text ] http://www.drthrasher.org
Jack D. Thrasher, PhD toxicology@... Sam-1 Trust,
PO Box 874 Alto, New Mexico 88312 505-336-8312 fax 425-675-7379

http://www-hsc.usc.edu/~kilburn/
Kaye H. Kilburn, M.D. kilburn@...
University of Southern California
Keck School of Medicine
Environmental Sciences Laboratory
2025 Zonal Avenue, CSC 201, Los Angeles, California 90033
text Dec, 1997 "Chemical Brain Injury"

http://groups.yahoo.com/group/aspartameNM/message/925
aspartame puts formaldehyde adducts into tissues, Part 1/2
full text, Trocho & Alemany 6.26.98: Murray 12.22.2 rmforall

http://groups.yahoo.com/group/aspartameNM/message/926
aspartame puts formaldehyde adducts into tissues, Part 2/2
full text, Trocho & Alemany 6.26.98: Murray 12.22.2 rmforall

http://ww.presidiotex.com/barcelona/index.html
Trocho C, Pardo R, Rafecas I, Virgili J, Remesar X,
Fernandez-Lopez JA, Alemany M ["Trok-ho"]
Formaldehyde derived from dietary aspartame binds to tissue
components in vivo. Life Sci 1998 Jun 26; 63(5): 337-49.
Departament de Bioquimica i Biologia Molecular, Facultat de Biologia,
Universitat de Barcelona, Spain.
http://www.presidiotex.com/barcelona/index.html
Maria Alemany, PhD (male) alemany@...

http://groups.yahoo.com/group/aspartameNM/message/864
Murray: Butchko, Tephly, McMartin: Alemany: aspartame formaldehyde
adducts in rats 9.8.2 rmforall
Prof. Alemany vigorously affirms the validity of the Trocho study
against criticism:
Butchko, HH et al [24 authors], Aspartame: review of safety.
Regul. Toxicol. Pharmacol. 2002 April 1; 35 (2 Pt 2): S1-93, review
available for $35, [an industry paid organ]. Butchko:
"When all the research on aspartame, including evaluations in both the
premarketing and postmarketing periods, is examined as a whole, it is
clear that aspartame is safe, and there are no unresolved questions
regarding its safety under conditions of intended use."
[They repeatedly pass on the ageless industry deceit that the methanol
in fruits and vegetables is as as biochemically available as that in
aspartame-- see the 1984 rebuttal by Monte, below.]

http://groups.yahoo.com/group/aspartameNM/message/911
RTP ties to industry criticized by CSPI: Murray: 12.9.2 rmforall

http://groups.yahoo.com/group/aspartameNM/message/935
comet assay finds DNA damage from sucralose, cyclamate, saccharin in
mice: Sasaki YF & Tsuda S Aug 2002: Murray 1.1.3 rmforall

http://groups.yahoo.com/group/aspartameNM/message/934
24 recent formaldehyde toxicity [Comet assay] reports:
Murray 12.31.2 rmforall

http://groups.yahoo.com/group/aspartameNM/message/622
Rich Murray: Gold: Koehler: Walton: Van Den Eeden: Leon:
aspartame toxicity 6.4.1 rmforall

http://groups.yahoo.com/group/aspartameNM/message/623
Rich Murray: Simmons: Gold: Schiffman: Spiers:
aspartame toxicity 6.4.1 rmforall

http://groups.yahoo.com/group/aspartameNM/message/917
HERP ranking of carcinogens (formaldehyde is very high):
www.berkeley.edu: Murray 12.14.2 rmforall

http://groups.yahoo.com/group/aspartameNM/message/870
Aspartame: Methanol and the Public Interest 1984:
Monte: Murray 9.23.2 rmforall

Dr. Woodrow C. Monte Aspartame: methanol and the public health.
Journal of Applied Nutrition 1984; 36 (1): 42-54.
(62 references) Professsor of Food Science
Director of the Food Science and Nutrition Laboratory
Arizona State University, Tempe, Arizona 85287
6411 South River Drive #61 Tempe, Arizona 85283-3337
602-965-6938 woody.monte@...
The methanol from 2 L of diet soda, 5.6 12-oz cans, 20 mg/can, is
112 mg, 10% of the aspartame. The EPA limit for water is 7.8 mg daily
for methanol (wood alcohol), a deadly cumulative poison. Many users
drink 1-2 L daily. The reported symptoms are entirely consistent
with chronic methanol toxicity. (Fresh orange juice has 34 mg/L, but,
like all juices, has 16 times more ethanol, which strongly protects
against methanol.)

"Fruit and vegetables contain pectin with variable methyl ester
content. However, the human has no digestive enzymes for pectin (6, 25)
particularly the pectin esterase required
for its hydrolysis to methanol (26).

Fermentation in the gut may cause disappearance of pectin (6) but the
production of free methanol is not guaranteed by fermentation (3). In
fact, bacteria in the colon probably reduce methanol directly to formic
acid or carbon dioxide (6) (aspartame is completely absorbed before
reaching the colon). Heating of pectins has been shown to cause
virtually no demethoxylation; even temperatures of 120 deg C produced
only traces of methanol (3). Methanol evolved during cooking of high
pectin foods (7) has been accounted for in the volatile fraction during
boiling and is quickly lost to the atmosphere (49).
Entrapment of these volatiles probably accounts for the elevation in
methanol levels of certain fruit and vegetable products
during canning (31, 33)."

Recent research [see links at end of post] supports his focus on the
methanol to formaldehyde toxic process:

"The United States Environmental Protection Agency in their Multimedia
Environmental Goals for Environmental Assessment recommends a minimum
acute toxicity concentration of methanol in drinking water at 3.9 parts
per million, with a recommended limit of consumption below 7.8 mg/day
(8). This report clearly indicates that methanol:

"is considered a cumulative poison
due to the low rate of excretion once it is absorbed.
In the body, methanol is oxidized to formaldehyde and
formic acid; both of these metabolites are toxic." (8)....

Recently the toxic role of formaldehyde (in methanol toxicity) has been
questioned (34). No skeptic can overlook the fact that, metabolically,
formaldehyde must be formed as an intermediate to formic acid
production (54).

Formaldehyde has a high reactivity which may be why it
has not been found in humans or other primates during methanol
poisioning (59)....

If formaldehyde is produced from methanol and does have a reasonable
half life within certain cells in the poisoned organism the chronic
toxicological ramifications could be grave.

Formaldehyde is a known
carcinogen (57) producing squamous-cell carcinomas by inhalation
exposure in experimental animals (22). The available epidemiological
studies do not provide adequate data for assessing the carcinogenicity
of formaldehyde in man (22, 24, 57).

However, reaction of formaldehyde
with deoxyribonucleic acid (DNA) has resulted in irreversible
denaturation that could interfere with DNA replication and result in
mutation (37)...."

http://groups.yahoo.com/group/aspartameNM/message/936
flawed test for aspartame DNA damage: Jeffrey & Williams 2000: Murray:
1.3.3 rmforall

http://groups.yahoo.com/group/aspartameNM/message/886
aspartame reduces aggression in rats?: Goerss, Wagner, Hill: Aug 2001:
Murray 11.10.2 rmforall
************************************************************************