RoomForAll: healthy and safe diet (aspartame)

methanol impurity in alcohol drinks [ and aspartame ] is turned into neurotoxic formic acid, prevented by folic acid, re Fetal Alcohol Syndrome,
BM Kapur, DC Lehotay, PL Carlen at U. Toronto, Alc Clin Exp Res 2007 Dec.
plain text: detailed biochemistry, CL Nie et al. 2007.07.18: Rich Murray
2008.02.24
http://rmforall.blogspot.com/2008_02_01_archive.htm
Sunday, February 24, 2008
http://groups.yahoo.com/group/aspartameNM/message/1524
____________________________________________________


[ Rich Murray comments: As a medical layman volunteer information
activist for aspartame and related toxicity issues since January 1999,
I note with appreciation the remarkable exponential progress on all
fronts, including a rapidly emerging consensus about the primary
importance of all toxicity challenges for our world.

This lengthy review features in detail two quite different, revolutionary
contributions, from Canada, and England and China.

It is indicative of our times that the CL Nie et al. study, 2007
appears in a free, open access journal-- indeed,
as all life and death information must.

Following rather vigorously, indeed blindly, the imperatives of
single-minded, profit-driven capitalist competition -- manipulating
adroitly research, education, media, citizens, governments -- many
great global corporations have inevitably created results that
oppose the common good. Alcohol and tobacco are well known.

Realistically, any further manipulations can only lead to inevitable
and even sudden corporate meltdowns, in the context of an
unfettered, cooperative, democratic global information forum,
the Internet.

Now, it is as easy and cheap to compose and instantly post a
30-page review as 3 pages a decade ago -- and such reviews
are archived forever in multiple collections, open via global search
engines to a billion Net citizens.

Perforce, and increasingly happily, all societal entities will have to
operate by high and shared voluntary universal standards
for the common good. ]


http://www.blackwell-synergy.com/doi/abs/10.1111/j.1530-0277.2007.00541.x

Alcoholism: Clinical and Experimental Research
Volume 31 Issue 12 Page 2114-2120, December 2007

Bhushan M. Kapur, b.kapur@utoronto.ca;
Arthur C. Vandenbroucke, PhD, FCACB
Yana Adamchik,
Denis C. Lehotay, dlehotay@health.gov.sk.ca;
Peter L. Carlen carlen@uhnres.utoronto.ca;
(2007) Formic Acid, a Novel Metabolite of Chronic Ethanol
Abuse, Causes Neurotoxicity, Which Is Prevented by Folic Acid
Alcoholism: Clinical and Experimental Research 31 (12), 2114-2120.
doi:10.1111/j.1530-0277.2007.00541.x

Abstract

Background:
Methanol is endogenously formed in the brain and is present as a
congener in most alcoholic beverages.

Because ethanol is preferentially metabolized over methanol (MeOH)
by alcohol dehydrogenase, it is not surprising that MeOH
accumulates in the alcohol-abusing population.

This suggests that the alcohol-drinking population will have higher
levels of MeOH's neurotoxic metabolite, formic acid (FA).

FA elimination is mediated by folic acid.

Neurotoxicity is a common result of chronic alcoholism.

This study shows for the first time that FA,
found in chronic alcoholics, is neurotoxic
and this toxicity can be mitigated by folic acid administration.

Objective:
To determine if FA levels are higher in the alcohol-drinking
population and to assess its neurotoxicity in organotypic
hippocampal rat brain slice cultures.

Methods:
Serum and CSF FA was measured in samples from both ethanol
abusing and control patients, who presented to a hospital emergency
department. [ CSF = Cerebral Spinal Fluid ]

FA's neurotoxicity and its reversibility by folic acid were assessed
using organotypic rat brain hippocampal slice cultures using clinically
relevant concentrations.

Results:
Serum FA levels in the alcoholics
(mean ± SE: 0.416 +- 0.093 mmol/l, n = 23)
were significantly higher than in controls
(mean ± SE: 0.154 +- 0.009 mmol/l, n = 82) (p < 0.0002).

FA was not detected in the controls' CSF (n = 20),
whereas it was >0.15 mmol/l in CSF of 3 of the 4 alcoholic cases.

Low doses of FA from 1 to 5 mmol/l added for 24, 48 or 72 hours
to the rat brain slice cultures caused neuronal death as measured by
propidium iodide staining.

When folic acid (1 umol/l) was added with the FA,
neuronal death was prevented. [ umol = micromole ]

Conclusions:
Formic acid may be a significant factor in the neurotoxicity of
ethanol abuse.

This neurotoxicity can be mitigated by folic acid administration
at a clinically relevant dose.

Key Words:
Formic Acid, Folic Acid, Methanol, Neurotoxicity, Alcoholism.

From the Department of Clinical Pathology (BMK),
Sunnybrook Health Science Centre,
Division of Clinical Pharmacology and Toxicology,
The Hospital for Sick Children, Toronto, Ontario, Canada;

St. Michael's Hospital (ACV), Toronto, Canada;

Department of Laboratory Medicine and Pathobiology
(BMK, ACV), Faculty of Medicine,
University of Toronto, Toronto, Ontario, Canada;

Departments of
Medicine (Neurology) and Physiology (YA, PLC),
Toronto Western Research Institute,
University of Toronto, Toronto, Ontario, Canada;

and University of Saskatchewan (DLC), Saskatchewan, Canada.

Received for publication May 1, 2007;
accepted September 24, 2007.

Reprint requests: Dr. Bhushan M. Kapur,
Department of Clinical Pathology,
Sunnybrook Health Science Centre,
2075 Bayview Ave, Toronto, Ontario, M4N 3M5, Canada;
Fax: 416-813-7562; E-mail: b.kapur@utoronto.ca;

Copyright 2007 by the Research Society on Alcoholism.
DOI: 10.1111/j.1530-0277.2007.00541.x
Alcoholism: Clinical and Experimental Research 2007 Dec.
Alcohol Clin Exp Res, Vol. 31, No 12, 2007: pp 2114-2120

NEUROTOXICITY AND BRAIN damage are common
concomitants findings of chronic alcoholism
(Carlen and Wilkinson, 1987; Carlen et al., 1981; Harper,
2007).

The cause of ethanol-induced neurotoxicity is still unclear.

We present here a novel hypothesis for neurotoxicity:
increased formic acid (FA) levels produced from methanol
(MeOH), whose catabolism is blocked by ethanol.

Axelrod and Daly (1965) demonstrated the endogenous formation
of MeOH from S-adenosylmethionine (SAM) in the pituitary
glands of humans and various other mammalian species.

Presence of MeOH in the breath of human subjects was
reported by Ericksen and Kulkarni (1963).

Most alcoholic beverages also have a small amount of MeOH
as a congener (Sprung et al., 1988).

As ethanol (EtOH) has a higher affinity for
alcohol dehydrogenase (ADH) than MeOH,
EtOH is preferentially metabolized (Mani et al., 1970).

As a result, MeOH accumulation from endogenously produced
MeOH, and/or, that consumed as part of an alcoholic beverage,
has been reported in concentrations up to 2 mmol/l in heavy
drinkers (Majchrowicz and Mendelson, 1971).

Toxicity resulting from MeOH consumption is extensively
documented in both humans and animals and has been
attributed to its metabolite, FA (Benton and Calhoun, 1952;
Roe, 1946, 1955; Wood, 1912; Wood and Buller, 1904).

The rate of formate oxidation and elimination is dependent on
adequate levels of hepatic folic acid, particularly hepatic
tetrahydrofolate (THF)
(Johlin et al., 1987; Tephly and McMartin, 1974).

Significantly higher formate levels were obtained when
folate-deficient animals were exposed to MeOH as compared
with folate-sufficient animals (Lee et al., 1994;
McMartin et al., 1975; Noker et al., 1980).

To understand ethanol's toxicity, one must consider FA
produced from MeOH, and its elimination mediated by folic acid.

We postulate that in the chronically drinking patient,
we will find higher levels of FA than in the nondrinking population,
and that formate is neurotoxic.

We also hypothesize that treatment with folic acid, which is a
critical factor in the catabolism of FA, can prevent or
diminish FA neurotoxicity.

METHODS

Patient Samples

During our study period of 4 months, 23 patients whose serum
showed the presence of both ethanol and trace amounts
(<2 mmol/l) of MeOH presented themselves to our emergency
department.

During the same period 4 patients, who were positive
for EtOH at admission, were admitted to the hospital.

During their stay, we received multiple samples as part
of their clinical follow-up.

We also received CSF and serum samples from 4 other patients
who were admitted in the hospital for alcohol abuse during our
study period.

All samples were analyzed for EtOH, MeOH, and FA.

As controls, we analyzed randomly the received serum (n = 82)
and CSF (n = 20) samples from inpatients, who did not have
any alcohol present at the time of admission.

All serum and CSF samples were collected as part of the patients'
clinical evaluation or follow-up.

Ethanol and MeOH were analyzed using headspace gas
chromatographic procedure.

FA was also analyzed by headspace gas chromatography
(Abolin et al., 1980).

Lower limits of detection for EtOH, MeOH, and FA
were 0.4, 0.8, and 0.13 mmol/l, respectively.

Organotypic Brain Slice Cultures

To study the neurotoxicity of FA, 2 sets of experiments using
organotypic brain slice cultures were performed:

(i) FA at concentrations of 1, 2 and 5 mmol/l was added to
organotypic hippocampal rat brain slice cultures
(n = 7 for each concentration).

To a second set of rat brain slice cultures, both FA,
at the above-mentioned concentrations, and 1 umol/l of folic acid
were also added.

Control brain slices, with and without folic acid, were also
processed with the experimental slices.

The time course and extent of cell death were determined by
measuring the fluorescence of the viability indicator,
propidium iodide (PI), at 24, 48 and 72 hours
after the application of FA alone
and in combination with folic acid.

Ensuring stable and reproducible measures of damage following
FA administration depended critically on the tissue culture
conditions of the hippocampal slice.

We noted that cultures which showed evident cell death before
experimental manipulations were more vulnerable to damage
from FA.

Hence, we were careful to use cultures that did not demonstrate
any apparent cell death.

The initial control images using PI were taken approximately 1 hour
before experimental measurements or manipulations.

Preparation of Organotypic Slice Cultures

Techniques for culturing brain slices have been described in detail
by Stoppini et al. (1991).
Briefly, the brains of 7-day-old male Wistar rats were aseptically
removed and immersed in ice-cold dissecting medium at pH 7.15
containing 50% minimum essential medium (MEM)
with no bicarbonate, 50% calcium and magnesium-free
Hanks balanced salt solution, 20 mM HEPES and 7.5 g/l d-glucose.
Hippocampi were dissected and coronal sections, 400-um thick,
were obtained.
Slices were transferred to a dish containing dissecting medium,
at room temperature.
The slices were then carefully separated
and transferred to sterile, porous membrane units with 0.4-um
diameter pores (Millicell-CM).
The membrane units were placed into 6-well trays,
each well containing 1 ml of culture medium, which is composed
of 50% MEM with Earl's salts, 2 mM l-glutamine, 25%
Earl's balanced salt solution, 25% normal horse serum,
6.5 g/l d-glucose, 20 mM HEPES buffer
and 50 mg/ml streptomycin-penicillin.
The pH of the medium was adjusted to 7.2 with HEPES buffer.

Cultures were kept in a tissue culture incubator for 2 weeks
at 36.8°C in 5% CO2
before the beginning of the experiments,
and fed 2 times a week by a 50% exchange of medium.

Assessment of Cell Death and Fluorescence Microscopy

Propidium iodide was applied to each dish at 10 mM,
30 minutes prior to the toxicity assessment.

PI fluorescence emission was measured immediately before,
at 24, 48 and 72 hours after the administration of FA
with a 4-X objective, using a confocal microscope
(BioRad, Hercules, CA).
A rhodamine filter (510 to 590 nm) was used to visualize PI
fluorescence emission. [ nm = nanometer ]
Gains and black levels were standardized for each experiment.
Fluorescence images were acquired and analyzed with the
Comos and the Confocal Assistant software packages (Bio-Rad).
Pixel intensity was measured either for the whole slice
or in selected areas of the hippocampus, CA1, CA3,
and dentate gyrus (DG), using a standard sized box.

At the end of each experiment, slices were killed by incubating
for 48 hours at 4°C in the presence of PI (Fig. 3).

The final PI fluorescence obtained after this treatment was
considered to be the fluorescence that closely represents
100% cellular death.

Cell death was then expressed as a percentage of the final
fluorescence minus the background fluorescence
taken before experiments.

The statistical comparisons between the control and injured groups
were performed using the unpaired Student's t-test.

Numerical values are expressed in the figures as mean
and standard error of mean.

Slices exhibiting PI staining before experiments or those revealing
any incomplete or absent hippocampal layers were excluded from the
assessment.

RESULTS

Human Subjects

Serum FA levels were significantly higher in the ethanol
positive patients when compared with the alcohol negative
controls (0.42 vs. 0.15 mmol/l; p < 0.0002) (Table 1).

In all the sequentially received samples from 4 inpatients,
EtOH declined linearly following zero-order kinetics.

Figure 1 shows the profile of 1 of these patients.

Both MeOH and FA levels remained almost constant
for a considerable period of time
and appeared to be at equilibrium.

In the final sample of all these patients,
MeOH levels had declined to almost 0,
whereas FA levels had risen exponentially (Table 2, Fig. 1).

This pattern was consistent in all patients.

The CSF and serum from 4 different alcohol-abusing patients
had FA in all 4 serum samples and
FA in 3 of the 4 CSF samples (Table 3).

In the CSF of nonethanol drinking control patients,
EtOH, MeOH, and FA were all
below the detection limit of the assays.

Organotypic Hippocampal Brain Slices Cultures Incubated
With FA

Formic acid from 1 to 5 mmol/l added for 72 hours caused
neuronal death as measured by PI staining (Figs 2 and 4).

A dose-response relationship was also observed (Fig. 2)
(p < 0.01).

When 1 uM folic acid was added to these slice cultures
along with the FA, neuronal death, secondary to FA,
was prevented (p = NS as compared
with control slice cultures with folic acid).

The effect was more pronounced at
48 hours than at 24 hours,
when compared with controls slice cultures with no folic acid.

Figure 3 (controls) shows with PI staining that there was
minimal cell death after 72 hours in cultures.

Figure 4 illustrates the dose and time response of FA neurotoxicity,
which affected the CA1 neuronal layer more
than the dentate granule cell layer.

It also shows the neuro-protective effect of 1 uM folic acid.


Table 1. Formic Acid Levels in Alcohol Positive
and Randomly Collected Samples
------------------------------- n --- Mean +- SE (mmol/l)

Control serum from
nonalcoholics---------------- 82 --- 0.154 --- 0.009

Alcohol-positive serum ------ 23 --- 0.416 --- 0.093
p < 0.0002


Table 2. Examples of Formic Acid Profile
in Ethanol-Positive Patients [ serum ]

[ To simplify, the highest levels were in Patient no. 4:
whose admission serum samples
and last serum samples at 30 h had:

Methanol --- 1.1 and ND mmol/l

Formic acid 0.25 and 1.95 mmol/l

ND = not detected

So, there is no data about the specific levels of
Formic acid in vulnerable tissues, like brain and eye.

However, it is clear that 30 hours after alcohol use,
all ethanol and methanol are gone from the blood serum,
while formic acid can be as high as 2 mmol/l. ]


Table 3. CSF and Serum
[ in 4 patients admitted for alcohol abuse.

At admission for alcohol abuse,
Patient no. 6 had in serum, methanol 1.7 mmol/l
and in CSF NSQ, Not Sufficient Quantity available for analysis,
and in serum, formic acid 2.25
and in CSF 0.7. ]


Fig. 1. Ethanol, methanol, and formic acid elimination profile
in an alcoholic during his stay in the hospital.


Fig. 2. Rat brain hippocampal slice cultures.
Control = no formic or folic acid;
control folic acid = 1 lmol folic acid only.
Data represented are means ± standard error of 7 slice cultures.
*p < 0.01 when compared with
controls at the corresponding time.
p = NS when 2 mmol formic + 1 umol folic acid
compared with control folic acid.


Fig. 3. Hippocampal slice cultures.
Intact controls: images at 24, 48 (B),
and 72 hours (C) and
killed slice cultures (D, 48 hours at 4 deg C).
(propidium iodide stains dead cells white).


Fig. 4. Hippocampal slice cultures:
Damage by formic acid is both dose and time dependent
and protection by folic acid (1 umol).
This figure shows effect of dose, time
and the neuro-protective effect provided by folic acid.
Hippocampal slice cultures treated
with 1, 2 and 5 mmol/ L of formic acid in
the presence and absence of folic acid (1 umol).
Images at 48 hours with
and without folic acid. (A, B)
Formic acid, 1 mmol; (C, D)
formic acid, 2 mmol; (E, F)
formic acid, 5 mmol;
protected slices, B, D, and F.
(Propidium iodide stains dead cells white).


DISCUSSION

There are at least 2 sources of MeOH:
endogenous production of MeOH (Axelrod and Daly, 1965;
Ericksen and Kulkarni, 1963; Gilg et al., 1987;
Iffland and Staak, 1990; Jones and Lowinger, 1988;
Majchrowicz and Mendelson, 1971; Roine et al., 1989;
Sarkola and Eriksson, 2001; Sprung et al., 1988),

and its presence as a congener in most alcoholic beverages
(Sprung et al., 1988).

MeOH concentrations between 4 and 4500 mg/l can be
present in various alcoholic beverages (Sprung et al., 1988).

Majchrowicz and Mendelson (1971) in an elegant experiment,
showed a rise in MeOH levels in subjects
drinking MeOH-free alcohol, thus supporting
the previous findings of endogenous production of MeOH.

Endogenous production of MeOH was described again in
2001 by Sarkola and Eriksson (2001).

These authors gave 4-methyl pyrazole,
a competitive inhibitor of ADH,
to volunteers not exposed to EtOH and observed a significant
elevation in endogenous EtOH and MeOH plasma levels.

MeOH levels rose linearly from 20 ± 14 umol/l to 39 ± 22 umol/l.

It took 195 minutes for EtOH levels to reach their peak (from
<5 umol/l to 30 ± 20 umol/l) concentrations as compared
with 420 minutes for MeOH,
suggesting gradual accumulation of MeOH
and preferential elimination of EtOH.

Altered pharmacokinetic behavior of MeOH in the presence of
EtOH has been demonstrated by various authors
(Lesch et al., 1990; Martensson et al., 1988).

As a result of continuous drinking
and the preferential metabolism of EtOH,
MeOH levels will rise in chronic drinkers
(Gilg et al., 1987; Iffland and Staak, 1990;
Jones and Lowinger, 1988; Majchrowicz and Mendelson, 1971;
Roine et al., 1989; Sprung et al., 1988).

MeOH has even been suggested as a marker for alcohol abuse
(Iffland and Staak, 1990; Roine et al., 1989).

As MeOH is metabolized to FA, this would suggest
that there could be a steady increase in FA levels
to some concentration at which equilibrium is reached.

It has been suggested that the concentration of MeOH
remains almost constant until EtOH levels have decreased to
about 4 mmol/l (Martensson et al., 1988).

Our data do indeed show this pattern.

In the 4 patients in whom we had multiple samples,
initially there was equilibrium between MeOH and FA.

The frequency of sample collection in all our patients
was based on the attending physician's clinical reason.

As a result, in all the 4 patients and the patient represented in
Fig. 1, there is a large time gap between the last 2 samples.

Our patient data (Fig. 1) do suggest that there must have been
an exponential rise in FA as EtOH approached 4 mmol/l
(Table 2).

Our data suggest that in the plasma of an alcohol-drinking person,
there can be elevated levels of FA (Table 3).

Two nonfree radical pathways have been proposed for formate
conversion to carbon dioxide: oxidation through the
catalase-peroxidative system (Chance, 1950),
and one-carbon pool.

Formate enters the one-carbon pool by combining with
THF to form 10-formyl-THF, a reaction catalyzed
by 10-formyl-THF synthetase (Johlin et al., 1987).

This is followed by the oxidation of 10-formyl-THF
to carbon dioxide mediated
by 10-formyl THF dehydrogenase (10-FTHFDH).

Studies have shown that this is the major route of formate
metabolism (Chiao and Stokstad, 1977; Johlin et al., 1987;
Makar and Tephly, 1976; Palese and Tephly, 1975)

and the predominant one in primates (McMartin et al., 1977).

Formate oxidation to carbon dioxide is dependent upon folic acid
in rats, monkeys (McMartin et al., 1977; Noker et al., 1980),
and in humans (liver) (Johlin et al., 1989).

Although liver is the main source for folate,
Neymeyer and Tephly (1994) and Neymeyer et al. (1997))
showed the presence of folate and 10-FTHFDH in the
retina, optic nerve, and in the various regions of the rat brain.

Folate was found to be between 3% and 14%
of that found in the liver.

The presence of folate and 10-FTHFDH in brain suggests
that formate can be metabolized in these tissues.

Folic acid deficiency is a common finding in chronic alcoholics,
(Eells et al., 2000; Halsted et al., 2002b; Herbert, 1990).

Chronic alcohol ingestion reduces the intestinal absorption of
dietary folic acid leading to a decrease in the folate metabolic
pool (Halsted et al., 2002b).

A decrease in this pool prolongs the formate blood levels
by decreasing the rate at which formate combines with THF,
the first step in its metabolism to carbon dioxide
and leads to formate-mediated cytotoxicity
(McMartin et al., 1977).

Folate deficiency can lead to a decrease in SAM
(Miller et al., 1994).

The overall status of the one-carbon pathway is also dependent
on the levels of methionine and vitamin B6 and B12
(Bailey and Gregory,1999; Barak et al., 1991;
Barber et al., 1999; Halsted et al., 2002a; Lucock, 2000;
Scott et al., 1993).

In situation of poor folate status, S-adenosylhomocysteine (SAH)
concentration increases due to the impairment of methyl group
synthesis and homocysteine re-methylation.

Inhibition by the resulting product, SAH, suppresses many of the
(SAM)-dependent methyl transferase reactions
(Selhub and Miller, 1992; Sokoro, 2007).

A number of studies have shown that there is enzymatic
activity in the brain which can metabolize both ethanol and
acetaldehyde (Brzezinski et al., 1999; Kapoor et al., 2006;
Roberto et al., 2006; Sun and Sun, 2001; Upadhya et al.,
2000; Vasiliou et al., 2006; Yadav et al., 2006;
Zimatkin et al., 2006).

Vasiliou et al. (2006) suggested that "Although the
contribution and CYP2E1 and catalase in ethanol oxidation
may be of little significance, these enzymes appear to play a
significant role in ethanol metabolism in the brain."

Patients in whom we had a CSF samples,
FA was present in 3 of the 4 patient's CSF.

Formic acid was present in all the 4 corresponding serum samples.

The presence of FA in the CSF suggests that either FA crosses
the blood-brain barrier or is formed in situ from the metabolism
of water-soluble MeOH that must have crossed
the blood-brain barrier.

Carlen et al. (1980) showed profound CSF anion gap metabolic
acidosis in alcoholic patients.

Our data showing the presence of FA in CSF may indeed explain
(Holt and Karty, 2003) the observed acidosis.

Formate can cause oxidative stress by producing free radicals
through the Fenton-like reaction (Dikalova et al., 2001;
Walling, 2007).

In this reaction, a hydroxyl radical (OH) is
formed through the Fenton-like reaction, which in turn
oxidized formate (HCO2),
forming the carbon dioxide anion radical (CO2).

The carbon dioxide anion radical then reacts
with molecular oxygen forming carbon dioxide and
the cytotoxic reactive oxygen species (ROS)- superoxide radical.


H2O2 + Fe,2+ --> *OH + Fe,3+ + OH,-

HCO2,- + *OH --> *CO2,- + H2O

*CO2,- + O2 --> CO2 + *O2,-


Chance has shown that formate can be metabolized by the
catalase-peroxidative system (Chance, 1950).

When anti-oxidants are depleted, increased ROS are formed
(Treichel et al., 2004).

Formic acid-induced cell damage has been attributed
to the generation of the cytotoxic ROS species.

FA disrupts mitochondrial electron transport and energy production
by inhibiting cytochrome oxidase activity (Nicholls, 1975, 1976;
Sharpe et al., 1982)
and causes cell death by increased production of cytotoxic ROS
secondary to the blockade of the electron transport chain
(Reed and Savage, 1995).

Formyl group (CHO) is transferred to THF
resulting in the formation of carbon dioxide and water
Makar et al., 1990; Medinsky et al., 1997).

Our organotypic brain slice studies suggest that there is a
dose and time relationship between FA and neuronal cell death.

FA levels achieved in the blood of the alcohol drinking
population can cause neuronal cell death.

The FA concentrations we used in our studies are representative
and were achieved in 2 of the 4 patients in whom we had sequential
samples.

It is remarkable that neuronal cell death could be prevented
by folic acid, although the mechanism of this protection is unknown.

There is a large body of literature relating folic acid deficiency
to neural tube defect, but, there are no references
relating low levels of FA to neurotoxicity.

There are a few studies relating FA and mitochondrial inhibition,
with MeOH intoxication and retinal damage
(Seme et al., 1999, 2001).

Another study demonstrated toxic effects of high concentrations
of formate in dissociated primary mouse neural cell cultures
(Dorman et al., 1993).

The concentration of formate that resulted
in 50% lactate dehydrogenase leakage after an 8-hour incubation
was estimated to be 45 mmol/l.

The total intracellular ATP concentration was significantly
decreased following either 20 or 40 mmol/l FA
exposure for 8 hour.

This is consistent with the hypothesis that FA may inhibit
mitochondrial function resulting in decreased intracellular ATP
and formate-induced neurotoxicity.

Using organotypic hippocampal slices, which preserve neuronal
circuitry and are easily accessible for experimental manipulations
(Stoppini et al., 1991),
our group has previously shown that
free radical overproduction in hippocampal pyramidal neurons
during ischemia/reoxygenation
depended on the activation of glutamate receptors,
and was associated with elevations of intracellular calcium.

Mitochondria are thought to be the principal source of
glutamate-mediated, calcium-dependent free radical production
in cultured cortical neurons
(Dugan et al., 1995; Reynolds and Hastings, 1995).

Although we did not investigate FA levels below 1 mmol/l,
it is conceivable that a continuous exposure to low,
but, above normal levels (>0.15 mmol/l), may also be cytotoxic
and may be part of the pathology of alcohol-related
organ damage (Jiang et al., 2003)
including the fetal alcohol spectrum disorder.

CONCLUSION

Our studies, for the first time, have shown that MeOH from
endogenous sources and from congeners present in alcoholic
beverages can lead to FA concentrations that are neurotoxic.

Therapeutic intervention with folic acid could be a significant
treatment modality in preventing FA mediated cytotoxicity,
especially neurotoxicity, in alcoholics.

ACKNOWLEDGMENT

This study was supported by a grant from the CIHR.

REFERENCES

Abolin C, McRae JD, Tozer TN, Takki S (1980)
Gas chromatographic head-space assay of formic acid
as methyl formate in biologic fluids:
potential application to methanol poisoning.
Biochem Med 23: 209-218.

Axelrod J, Daly J (1965)
Pituitary gland: enzymic formation of methanol from
5-adenosyl-methionine.
Science 150: 892-893.

Bailey LB, Gregory JF III (1999)
Folate metabolism and requirements.
[Review] [36 refs]. J Nutr 129: 779-782.

Barak AJ, Beckenhauer HC, Tuma DJ (1991)
Hepatic transmethylation and blood alcohol levels.
Alcohol Alcohol 26: 125-128.

Barber RC, Lammer EJ, Shaw GM, Greer KA, Finnell RH (1999)
The role of folate transport and metabolism in neural tube defect risk.
Mol Genet Metab 66: 1-9.

Benton CD, Calhoun FP (1952)
The ocular effects of methyl alcohol poisoning.
Report of a catastrophe involving three hundred and twenty persons.
Trans Am Acad Opthalmol 56: 875-883.

Brzezinski MR, Boutelet-Bochan H, Person RE, Fantel AG,
Juchau MR (1999)
Catalytic activity and quantitation of cytochrome P-450 2E1
in prenatal human brain.
J Pharmacol Exp Ther 289: 1648-1653.

Carlen PL, Kapur B, Huszar LA, Lee MA, Moddel G, Singh R,
Wilkinson DA (1980)
Prolonged cerebrospinal fluid acidosis in recently abstinent
chronic alcoholics.
Neurology 30: 956-962.

Carlen PL, Wilkinson DA (1987)
Reversibility of alcohol-related brain damage:
clinical and experimental observations.
Acta Medica Scandinavica Supplementum 717: 19-26.

Carlen PL, Wilkinson DA, Wortzman G, Holgate R, Cordingley J,
Lee MA, Huszar L, Moddel G, Singh R, Kiraly L, Rankin JG (1981)
Cerebral atrophy and functional deficits in alcoholics
without clinically apparent liver disease.
Neurology 31: 377-385.

Chance B (1950)
On the reaction of catalase peroxide with acceptors.
J Biol Chem 182: 649-658.

Chiao F, Stokstad EL (1977)
Effect of methionine on the metabolism of formate and histidine
by rats fed folate/ vitamin B-12-methionine-deficient diet.
Biochim Biophys Acta 497: 225-233.

Dikalova AE, Kadiiska MB, Mason RP (2001)
An in vivo ESR spin-trapping study: free radical generation in rats
from formate intoxication -- role of the Fenton reaction.
Proc Natl Acad Sci U S A 98: 13549-13553.

Dorman DC, Bolon B, Morgan KT (1993)
The toxic effects of formate
in dissociated primary mouse neural cell cultures.
Toxicol Appl Pharmacol 122: 265-272.

Dugan LL, Sensi SL, Canzoniero LM, Handran SD, Rothman SM,
Lin TS, Goldberg MP, Choi DW (1995)
Mitochondrial production of reactive oxygen species
in cortical neurons following exposure to N-methyl-D-aspartate.
J Neurosci 15: 6377-6388.

Eells JT, Gonzalez-Quevedo A, Santiesteban FR, McMartin KE,
Sadun AA (2000)
[Folic acid deficiency and increased concentrations of formate in
serum and cerebrospinal fluid of patients
with epidemic optical neuropathy].
[Spanish].
Revista Cubana de Medicina Tropical 52: 21-23.

Ericksen SP, Kulkarni AB (1963)
Methanol in normal breath.
Science 141: 639-640.

Gilg T, von Meyer L, Liebhardt E (1987)
[Formation and accumulation of endogenous methanol
in relation to alcohol burden]. [German].
Blutalkohol 24: 321-332.

Halsted CH, Villanueva JA, Devlin AM (2002a)
Folate deficiency, methionine metabolism, and alcoholic liver disease.
Alcohol 27: 169-172.

Halsted CH, Villanueva JA, Devlin AM, Chandler CJ (2002b)
Metabolic interactions of alcohol and folate.
J Nutr 132(8 Suppl): 2367S-2372S.

Harper C (2007)
The neurotoxicity of alcohol. [Review] [65 refs].
Hum Exp Toxicol 26: 251-257.

Herbert V (1990)
Development of human folate deficiency,
in Folic Acid Metabolism in Health and Disease
(Picciano MF, Skostad ELR, Gregory JF, Eds),
pp 195-210. Wiley, New York.

Holt J, Karty JM (2003)
Origin of the acidity enhancement of formic acid over methanol:
resonance versus inductive effects.
J Am Chem Soc 125: 2797 - 2803.

Iffland R, Staak M (1990)
Methanol and isopropanol as markers of alcoholism. [German].
Beitr Gerichtl Med 48: 173-177.

Jiang R, Hu FB, Giovannucci EL, Rimm EB, Stampfer MJ,
Spiegelman D, Rosner BA, Willett WC (2003)
Joint association of alcohol and folate intake
with risk of major chronic disease in women.
Am J Epidemiol 158: 760-771.

Johlin FC, Fortman CS, Nghiem DD, Tephly TR (1987)
Studies on the role of folic acid and folate-dependent enzymes
in human methanol poisoning.
Mol Pharmacol 31: 557-561.

Johlin FC, Swain E, Smith C, Tephly TR (1989)
Studies on the mechanism of methanol poisoning:
purification and comparison of rat and human liver
10-formyltetrahydrofolate dehydrogenase.
Mol Pharmacol 35: 745-750.

Jones AW, Lowinger H (1988)
Relationship between the concentration of ethanol and methanol
in blood samples from Swedish drinking drivers.
Forensic Sci Int 37: 277-285.

Kapoor N, Pant AB, Dhawan A, Dwievedi UN, Gupta YK,
Seth PK, Parmar D (2006)
Differences in sensitivity of cultured rat brain neuronal and glial
cytochrome P450 2E1 to ethanol.
Life Sci 79: 1514-1522.

Lee EW, Garner CD, Terzo TS (1994)
Animal model for the study of methanol toxicity:
comparison of folate-reduced rat responses
with published monkey data.
J Toxicol Environ Health 41: 71-82.

Lesch OM, Kefer J, Lentner S, Mader R, Marx B, Musalek M,
Nimmerrichter A, Preinsberger H, Puchinger H,
Rustembegovic A (1990)
Diagnosis of chronic alcoholism -- classificatory problems.
Psychopathology 23: 88-96.

Lucock M (2000)
Folic acid: nutritional biochemistry, molecular biology, and
role in disease processes.
Mol Genet Metab 71: 121-138.

Majchrowicz E, Mendelson JH (1971)
Blood methanol concentrations during experimentally induced
ethanol intoxication in alcoholics.
J Pharmacol Exp Ther 179: 293-300.

Makar AB, Tephly TR (1976)
Methanol poisoning in the folate-deficient rat.
Nature 261: 715-716.

Makar AB, Tephly TR, Sahin G, Osweiler G (1990)
Formate metabolism in young swine.
Toxicol Appl Pharmacol 105: 315-320.

Mani JC, Pietruszko R, Theorell H (1970)
Methanol activity of alcohol dehydrogenases
from human liver, horse liver, and yeast.
Arch Biochem Biophys 140: 52-59.

Martensson E, Olofsson U, Heath A (1988)
Clinical and metabolic features of ethanol-methanol poisoning
in chronic alcoholics.
Lancet 1: 327 - 328.

McMartin KE, Makar AB, Martin A, Palese M, Tephly TR (1975)
Methanol poisoning
1. The role of formic acid in the development of metabolic
acidosis in the monkey and the reversal by 4-methylpyrazole.
Biochem Med 13: 319-333.

McMartin KE, Martin-Amat G, Makar AB, Tephly TR (1977)
Methanol poisoning.
V. Role of formate metabolism in the monkey.
J Pharmacol Exp Ther 201: 564-572.

Medinsky MA, Dorman DC, Bond JA, Moss OR, Janszen DB,
Everitt JI (1997)
Pharmacokinetics of methanol and formate in female cynomolgus
monkeys exposed to methanol vapors.
Res Rep Health Eff Inst 77: 1-30;
Discussion 31-38.

Miller JW, Nadeau MR, Smith J, Smith D, Selhub J (1994)
Folate-deficiency-induced homocysteinaemia in rats:
disruption of S-adenosylmethionine's co-ordinate regulation
of homocysteine metabolism.
Biochem J 298(Pt 2): 415-419.

Neymeyer VR, Tephly TR (1994)
Detection and quantification of
10-formyl-tetrahydrofolate dehydrogenase (10-FTHFDH)
in rat retina, optic nerve, and brain.
Life Sci 54: L395-L399.

Neymeyer V, Tephly TR, Miller MW (1997)
Folate and 10-formyltetrahydro-folate dehydrogenase (FDH)
expression in the central nervous system of the mature rat.
Brain Res 766: 195-204.

Nicholls P (1975)
Formate as an inhibitor of cytochrome C oxidase.
Biochem Biophys Res Commun 67: 610-616.

Nicholls P (1976)
The effect of formate on cytochrome aa3 and on
electron transport in the intact respiratory chain.
Biochim Biophys Acta 430: 13-29.

Noker PE, Eells JT, Tephly TR (1980)
Methanol toxicity: treatment with folic acid
and 5-formyl tetrahydrofolic acid.
Alcohol Clin Exp Res 4: 378-383.

Palese M, Tephly TR (1975)
Metabolism of formate in the rat.
J Toxicol Environ Health 1: 13-24.

Reed DJ, Savage MK (1995)
Influence of metabolic inhibitors on mitochondrial permeability
transition and glutathione status.
Biochim Biophys Acta 1271: 43-50.

Reynolds IJ, Hastings TG (1995)
Glutamate induces the production of reactive oxygen species
in cultured forebrain neurons following NMDA receptor
activation.
J Neurosci 15(Pt 1): 3318-3327.

Roberto M, Treistman SN, Pietrzykowski AZ, Weiner J, Galindo R,
Mameli M, Valenzuela F, Zhu PJ, Lovinger D, Zhang TA,
Hendricson AH, Morrisett R, Siggins GR (2006)
Actions of acute and chronic ethanol on presynaptic terminals.
Alcohol Clin Exp Res 30: 222-232.

Roe O (1946)
Methanol poisoning its clinical course, pathogenesis and treatment.
Acta Medica Scand 126(Suppl.):182.

Roe O (1955)
The metabolism and toxicity of methanol.
Pharmacol Rev 7: 399-412.

Roine RP, Eriksson CJ, Ylikahri R, Penttila A, Salaspuro M (1989)
Methanol as a marker of alcohol abuse.
Alcoholism 13: 172-175.

Sarkola T, Eriksson CJ (2001)
Effect of 4-methylpyrazole on endogenous
plasma ethanol and methanol levels in humans.
Alcohol Clin Exp Res 25: 513-516.

Scott JM, McPartlin J, Molloy A, McNulty H, Halligan A,
Darling M, Weir DG (1993)
Folate metabolism in pregnancy.
Adv Exp Med Biol 338: 727 - 732.

Selhub J, Miller JW (1992)
The pathogenesis of homocysteinemia: interruption of the
coordinate regulation by S-adenosylmethionine of the
remethylation and transsulfuration of homocysteine.
Am J Clin Nutr 55: 131-138.

Seme MT, Summerfelt P, Henry MM, Neitz J, Eells JT (1999)
Formate-induced inhibition of photoreceptor function
in methanol intoxication.
J Pharmacol Exp Ther 289: 361-370.

Seme MT, Summerfelt P, Neitz J, Eells JT, Henry MM (2001)
Differential recovery of retinal function after mitochondrial inhibition
by methanol intoxication.
Invest Ophthalmol Vis Sci 42: 834-841.

Sharpe JA, Hostovsky M, Bilbao JM, Rewcastle NB (1982)
Methanol optic neuropathy: a histopathological study.
Neurology 32: 1093-1100.

Sokoro A (2007)
The Role of Folate Status in Formate Metabolism and Its
Relationship to Antioxidant Capacity During Alcohol Intoxication.
(Ph.D thesis). University of Saskatchewan, Saskatchewan, Canada.

Sprung R, Bonte W, Lesch OM (1988)
Methanol -- an up-to-now neglected constituent
of all alcoholic beverages. A new biochemical approach
to the problem of chronic alcoholism. [German].
Wien Klin Wochenschr 100: 282 - 288.

Stoppini L, Buchs PA, Muller D (1991)
A simple method for organotypic cultures of nervous tissue.
J Neurosci Methods 37: 173-182.

Sun AY, Sun GY (2001)
Ethanol and oxidative mechanisms in the brain.
J Biomed Sci 8: 37-43.

Tephly TR, McMartin KE (1974)
Methanol metabolism and toxicity,
in Aspartame: Physiology and Biochemistry
(Stegink LD, Filer LJ Jr eds),
pp 111-140. Markel Deccer, New York/Basel.

Treichel JL, Henry MM, Skumatz CM, Eells JT, Burke JM (2004)
Antioxidants and ocular cell type differences in cytoprotection
from formic acid toxicity in vitro.
Toxicol Sci 82: 183-192.

Upadhya SC, Tirumalai PS, Boyd MR, Mori T,
Ravindranath V (2000)
Cytochrome P4502E (CYP2E) in brain: constitutive expression,
induction by ethanol and localization
by fluorescence in situ hybridization.
Arch Biochem Biophys 373: 23-34.

Vasiliou V, Ziegler TL, Bludeau P, Petersen DR, Gonzalez FJ,
Deitrich RA (2006)
CYP2E1 and catalase influence ethanol sensitivity
in the central nervous system.
Pharmacogenet Genomics 16: 51-58.

Walling C (2007)
Fenton's reagent revisited.
Acc Chem Res 8: 125-131.

Wood CH (1912)
Death and blindness from methyl or wood alcohol poisoning.
JAMA 59: 1962-1969.

Wood CA, Buller F (1904)
Poisoning by wood alcohol: Cases of death and blindness
from Columbian sprits and other methylated preparations.
JAMA 43: 972-977.

Yadav S, Dhawan A, Singh RL, Seth PK, Parmar D (2006)
Expression of constitutive and inducible cytochrome P450 2E1
in rat brain.
Mol Cell Bio-chem 286: 171-180.

Zimatkin SM, Pronko SP, Vasiliou V, Gonzalez FJ,
Deitrich RA (2006) Enzymatic mechanisms of ethanol oxidation
in the brain.
Alcohol Clin Exp Res 30: 1500-1505.
____________________________________________________



folic acid prevents neurotoxicity from formic acid, made by body
from methanol impurity in alcohol drinks [ also 11 % of aspartame ],
BM Kapur, PL Carlen, DC Lehotay, AC Vandenbroucke,
Y Adamchik, U. of Toronto, 2007 Dec., Alcoholism Cl. Exp. Res.:
Murray 2007.11.27
http://rmforall.blogspot.com/2007_11_01_archive.htm
Wednesday, November 27, 2007
http://groups.yahoo.com/group/aspartameNM/message/1495


http://www.faslink.org/Formic%20Acid%20Kapur.htm

Brief Summary:

Methanol in small amounts is present along with ethanol in beverage
alcohol.
[Murray: and about the same amounts from aspartame diet sodas]

The body's natural enzymes preferentially metabolize ethanol while
methanol breaks down into highly neurotoxic Formic Acid.

Use of high levels of Folic Acid was found to inhibit brain damage
caused by the methanol.

The use of Folic Acid during pregnancy has been recommended
for several years to prevent neural tube defects.

However, this study indicates that even higher levels of Folic Acid
can be very beneficial to the developing baby, particularly where
alcohol exposure is a factor.

Folic Acid is mandated as an additive to all flour sold in Canada.

The debate has begun on its required addition to all beverage
alcohol to help mitigate damage caused to both infants and adults.


Formic Acid in the Drinking patient and the expectant mother
Dr. Bhushan M. Kapur
Departments of Laboratory Medicine,
St. Michael's Hospital , Toronto, Ontario, Canada

Abstract

Methanol is produced endogenously in the pituitary glands of humans
and is present as a congener in almost all alcoholic beverages.

Ethanol and methanol are both bio-transformed by alcohol
dehydrogenase; however, ethanol has greater affinity for the enzyme.

Since ethanol is preferentially metabolized by the enzyme, it is not
surprising that trace amounts of methanol, most likely originating from
both sources, have been reported in the blood of people
who drink alcohol.

Toxicity resulting from methanol is very well documented
in both humans and animals and is attributed to its toxic metabolite
formic acid.

To understand ethanol toxicity
and Fetal Alcohol Spectrum Disorders, it is important to consider
methanol and its metabolite, formic acid, as
potential contributors to the toxic effects of alcohol.

Accumulation of methanol suggests that alcohol-drinking
population should have higher than baseline levels of formic acid.

Our preliminary studies do indeed show this.

Chronic low-level exposure to methanol has been suggested to
impair human visual functions.

Formic acid is known to be toxic to the optic nerve.

Ophthalmological abnormalities are a common finding in children
whose mothers used alcohol during pregnancy.

Formic acid, a low molecular weight substance, either crosses the
placenta or may be formed in-situ from the water soluble methanol
that crosses the placenta.

Embryo toxicity from formic acid has been reported
in an animal model.

To assess neurotoxicity we applied low doses of formic acid
to rat brain hippocampal slice cultures.

We observed neuronal death with a time and dose response.

Formic acid requires folic acid as a cofactor for its elimination.

Animal studies have shown that when folate levels are low, the
elimination of formic acid is slower and formate levels are elevated.

When folic acid was added along with the formic acid
to the brain slice cultures, neuronal death was prevented.

Therefore, folate deficient chronic drinkers may be at higher risk of
organ damage.

Women who are folic acid deficient and consume alcohol may have
higher levels of formic acid and should they become pregnant,
their fetus may be at risk.

To our knowledge low level chronic exposure to formic acid and its
relationship to folic acid in men or women who drink alcohol has
never been studied.

Our hypothesis is that the continuous exposure to low levels of
formic acid is toxic to the fetus and may be part of the etiology of
Fetal Alcohol Spectrum Disorders.
____________________________________________________


http://www.come-over.to/FAS/

The incidence of Fetal Alcohol Syndrome in America
is 1.9 cases per 1,000 births (1/500).

Incidence of babies with disabilities
resulting from prenatal alcohol exposure: 1/100!
____________________________________________________


http://groups.yahoo.com/group/aspartameNM/message/1067
eyelid contact dermatitis by formaldehyde from aspartame,
AM Hill & DV Belsito, Nov 2003: Murray 4.4.4 rmforall [150 KB]

[ Extracts ]

McMartin, KE et al 1979, put 3,000 mg/kg methanol in the
stomachs of small monkeys and, 18 hours later found accumulation
of formate in liver, kidney, optic nerve, cerebrum, and midbrain
in 2 of three monkeys.

Biochemical Pharmcacology 1979: 28; 645-649.
Lack of a role for formaldehyde in methanol poisoning in the monkey.
Kenneth E. McMartin, Gladys Martin-Amat, Patricia E. Noker
and Thomas R. Tephly kmcmar@lsuhsc.edu;
The Toxicology Center, Dept. of Pharmacology,
University of Iowa, Iowa City, Iowa 52242

K.E. McMartin and T.R. Tephly, authors of many pro-aspartame
studies, in Biochemical Pharmacology (1979) remarked,
"It is now generally accepted
that the toxicity of methanol is due to the formation of toxic
metabolites, either formaldehyde or formic acid."

They put damage doses of methanol into the stomachs
of three monkeys,
and, using insensitive tests, found no formaldehyde in many tissues --
except for a single datum in the midbrain,
1.5 times their detection limit.

They did report widespread accumulation of formic acid
in five tissues.

The use of inadequate tests is common in industry research that is
funded to claim the safety of profitable toxins.

Since then, industry scientists have been very wary of doing studies
on primates, which all too easily show the dangers to humans.

"Abstract [ not given in PubMed ]:
[ My briefer comments are in square brackets. ]

Methanol was administered [ by nasogastric tube ] either to untreated
cynomolgus monkeys [ 2-3.5 kg ] or to a folate-deficient cynomolgus
monkey which exhibits exceptional sensitivity to the toxic effects of
methanol.

Marked formic acid accumulation in the blood and in body fluids and
tissues was observed.

No formaldehyde accumulation was observed in the blood and no
formaldehyde was detected in the urine, cerebrospinal fluid, vitreous
humor, liver, kidney, optic nerve, and brain in these monkeys at a
time when marked metabolic acidosis and other characteristics of
methanol poisoning were observed.

Following intravenous infusion into the monkey, formaldehyde was
rapidly eliminated from the blood with a half-life of about 1.5 min
and formic acid levels promptly increased in the blood.

Since formic acid accumulation accounted for the metabolic acidosis
and since ocular toxicity essentially identical to that produced in
methanol poisoning has been described after formate treatment,
the predominant role of formic acid as the major metabolic agent
for methanol toxicity is certified.

Also, results suggest that formaldehyde is not a major factor in the
toxic syndrome produced by methanol in the monkey."

"It is now generally accepted that the toxicity of methanol is due to
the formation of toxic metabolites (1,2),
either formaldehyde or formic acid."

So, this is an acute toxicity study, with little relevance for chronic
long-term, low-level exposure.

Monkeys, like people, are susceptible to methanol toxicity.

This team cites their six previous methanol in monkey studies,
from 1975 to 1977.

The report is difficult to understand, since the three monkeys were
treated differently, and different assays were used.

For the methanol sensitive, folate-deficient monkey A, the assay
used was the chromatropic acid method,
with a detection limit of .025 mmol/L.

None of the five tissues showed any formaldehyde with this assay,
except the midbrain, 0.14 mmol/kg wet weight tissue
[ units converted from their 0.14 micromole/gm -- just
1.5 times the detection limit of .09 mmol/kg wet tissue weight
(given on p. 648).
[ Since 1 kg of water is 1 L, 1 mmol/kg is equivalent to 1 mmol/L. ]

Meanwhile, in the methanol sensitive, folate-deficient monkey A,
the blood formate level rose by 18 hours from 0.18 to 10.02 mEq/L.
[ I assume that a mEq is equivalent to a mmol -- let me know
if I'm wrong. ]

The formate detection limits for the assays were not given
in this report.

The formate level in the vitreous humor of the eye of monkey A
was 7.90 mEq/L.

It is well known that formate is extremely damaging to the eye.

For unexplained reasons, formate levels in the five tissues and
cerebrospinal fluid were not measured in the methanol sensitive,
folate-deficient monkey A.,
in the cerebrospinal fluid of monkey B,
or in the optic nerve of monkey C.

Formaldehyde was not measured in the optic nerve of Monkey A.

The kidney formate level for monkey B was 6.33
and for C was only 0.44,
with no comment or explanation given.

The experiment seems arbitrary, capricious, and erratic.

For monkey A, after 18 hours, the urine formaldehyde level was
below detection level, while urine formate was 115.80 mEq/L -- so
much of the formaldehyde had been converted into formic acid,
another cumulative, potent toxin.

"In the presence of high formate values and definitive evidence of
toxicity in methanol-poisoned monkeys, no measurable formaldehyde
was found in the body tissues that were tested."

It is reasonable to surmise that more sensitive assays would have found
formaldehyde and formate bound to and reacted with a variety of cellular
substances in all tissues -- just as the 1998 Trocho study confirmed.
(Appendix E)

Monkeys B and C were normal, not extra vulnerable to methanol,
and were given 3,000 mg/kg methanol, and samples taken at 18 hr.

Formaldehyde was detected only in the blood of Monkey B,
while formate was found in 8 and 10, respectively,
of the 10 fluid and tissue samples in Monkeys B and C.

For instance, the lowest value of formate, except for zero-time blood,
for each monkey was in the midbrain, 2.16 mmol/kg for Monkey B
(24 times the detection limit for the chromatropic acid method)
and 1.02 mmol/kg (1.3 times the detection for the dimedon method)
for Monkey C.

This shows accumulation of formate in liver, kidney, optic nerve,
cerebrum, and midbrain.

"Thus, whereas one can associate formate intimately with ocular
toxicity in the monkey, no association of formaldehyde with ocular
toxicity can be made at this time.

It is not possible to completely eliminate formaldehyde as a toxic
intermediate because formaldehyde could be formed slowly within
cells and interfere with normal cellular function without ever obtaining
levels that were detectable in body fluids..."

"Acknowledgements-- This research was supported by
NIH grant GM 19420
and GM 12675." [not funded by the industry]


Life Sci 1991; 48(11): 1031-41.
The toxicity of methanol.
Tephly TR.
Department of Pharmacology, University of Iowa, Iowa City 52242.

"Abstract:
Methanol toxicity in humans and monkeys is characterized by a latent
period of many hours followed by a metabolic acidosis
and ocular toxicity.

This is not observed in most lower animals.

The metabolic acidosis and blindness is apparently due to
formic acid accumulation in humans and monkeys,
a feature not seen in lower animals.

The accumulation of formate is due to a deficiency in formate
metabolism which is, in turn, related, in part,
to low hepatic tetrahydrofolate (H4 folate).

An excellent correlation between hepatic H4 folate and
formate oxidation rates has been shown within and across species.

Thus, humans and monkeys possess low hepatic H4 folate levels,
low rates of formate oxidation and accumulation of formate
after methanol.

Formate, itself, produces blindness in monkeys in the absence of
metabolic acidosis.

In addition to low hepatic H4 folate concentrations, monkeys and
humans also have low hepatic 10-formyl H4 folate dehydrogenase
levels, the enzyme which is the ultimate catalyst for conversion of
formate to carbon dioxide.

This review presents the basis for the role of folic acid-dependent
reactions in the regulation of methanol toxicity.
Publication Types: Review Review, Academic PMID: 1997785"

p. 1035 "In the past, formaldehyde has often been suggested as the
methanol metabolite which produces toxicity (34,35).

Today, a great deal of information is available concerning its lack of
such a role.

The presence of elevated formaldehyde levels in body fluids or
tissues following methanol administration has not been observed.

No formaldehyde has been detected in blood, urine or tissues
obtained from methanol-treated animals (36,37) and,
in methanol-poisoned humans, formaldehyde increases
have not been observed....

About 85% of a low dose of 14C-formaldehyde [radioactive label]
is excreted as pulmonary 14CO2 (49,50)....."

[ This suggests that 15% of the formaldehyde is indeed retained in
the body, a very significant result, considering its extreme
and complex toxicity. ]

49. W.B. Neely, Biochem. Pharmacol. 13: 1137-1142 (1964).

50. Xenobiotica 1982 Feb; 12(2): 119-24.
Formaldehyde metabolism by the rat: a re-appraisal.
Mashford PM, Jones AR.
1. The metabolism of [14C]formaldehyde has been investigated
in the male Sprague-Dawley rat.
It is extensively oxidized to CO2 and formate,
which is excreted in the urine.
2. Two radioactive compounds isolated from the urine of rats dosed
with [14C] formaldehyde have been identified as
N-(hydroxymethyl)urea and
N,N'-bis-(hydroxymethyl)urea, and shown to be urinary artefacts.
3. Previous studies of the metabolism of formaldehyde by rats have
been re-appraised.
Differences in the rate of oxidation of formaldehyde in various strains
of rats result in the excretion of different urinary metabolites and, in
some cases, formaldehyde.
Excretion of formaldehyde leads to the formation of several artefacts
depending on the components present in the urine. PMID: 6806997
____________________________________________________


new details on how formaldehyde and formic acid from methanol are
neurotoxic: Chun Lai Nie, Rong Giao He, et al, PLoS ONE 2(7):
e629 2007.07.18 Chinese Academy of Sciences, Beijing:
Murray 2007.09.01
http://groups.yahoo.com/group/aspartameNM/message/1470

" Recent studies have shown that neurodegeneration
is closely related to misfolding and aggregation of neuronal tau. "

" The significant protein tau aggregation induced by formaldehyde
and the severe toxicity of the aggregated tau to neural cells may
suggest that toxicity of methanol and formaldehyde ingestion
is related to tau misfolding and aggregation. "

" Neuronal tau is an important protein in promoting and stabilizing
the microtubule system involved in cellular transport and neuronal
morphogenesis. "

" Both formaldehyde and acetaldehyde can go through the
blood-brain barrier and cause some lesions to CNS,
especially our visual system [38].

Clinically, the lethal dose of formaldehyde for human beings is
about 0.08% in the circulation [39].

We have shown in the present study that formaldehyde can
significantly induce tau aggregation and polymerization at
concentrations even lower than 0.08%,
the clinical dose of toxicosis. "

" Formaldehyde exposure leads to formation of DNA/protein
crosslinks, a major mechanism of DNA damage.

The DNA/protein crosslinks have been used as a measure
of dose in drug delivery [20].

Formaldehyde, as a crosslinking agent, also reacts with
thiol and amino groups, leading to protein polymerization [21], [22].

Furthermore, methanol ingestion is an important public health
concern because of the selective actions of its toxic metabolites,
formaldehyde and formic acid, on the retina, the optic nerves
and the central nervous system (CNS) [23].

Illicit consumption of industrial methylated spirits can cause severe
and even fatal illness [24].

In the liver and retina, methanol is oxidized by alcohol
dehydrogenase, resulting in formaldehyde.

In semicarbazide-sensitive amine oxidase (SSAO)-mediated
pathogenesis of Alzheimer's disease, formaldehyde interacts
with B-amyloids and produces irreversibly cross-linked neurotoxic
amyloid-like complexes [21], [22], [25].

We have examined the role of formaldehyde in misfolding
of protein tau [26].

In particular, we investigated the toxicity of formaldehyde-induced
tau aggregates on human neuroblastoma cells (SH-SY5Y cell line)
and rat hippocampal cells [27].

The results showed that low concentrations (0.01 - 0.1%) of
formaldehyde are sufficient to induce formation of amyloid-like tau
aggregates, which can induce apoptosis of both SH-SY5Y
and hippocampal cells.

This may be significant to understand the mechanism of chronic
damage caused by methanol toxicity
and formaldehyde stress [18], [28].

However, we have still not known the mechanism of protein tau
aggregation in the presence of formaldehyde at low concentrations.

The present study concerns the characteristic of misfolding and
polymerization of extracellular and intracellular neuronal tau induced
by formaldehyde at low concentrations. "

http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17637844
http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000629
free full text

Formaldehyde at Low Concentration Induces Protein Tau
into Globular Amyloid-Like Aggregates In Vitro and In Vivo
PLoS ONE. 2007 Jul 18; 2(7): e629.
doi:10.1371/journal.pone.0000629
Chun Lai Nie 1,
Yan Wei 1,
Xinyong Chen 2,
Yan Ying Liu 1,
Wen Dui 1,
Ying Liu 1,
Martyn C. Davies 2, Martyn.Davies@nottingham.ac.uk;
Saul J.B. Tendler 2, Saul.Tendler@nottingham.ac.uk;
Rong Giao He 1* herq@sun5.ibp.ac.cn;

1 State Key Laboratory of Brain and Cognitive Science,
Institute of Biophysics, Graduate School,
Chinese Academy of Sciences, Chaoyang District, Beijing, China,

2 Laboratory of Biophysics and Surface Analysis,
School of Pharmacy, The University of Nottingham,
Nottingham, United Kingdom

Received: March 5, 2007; Accepted: June 13, 2007;
Published: July 18, 2007

Copyright: © 2007 Nie et al.
This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which
permits unrestricted use, distribution, and reproduction in any
medium, provided the original author and source are credited.

* To whom correspondence should be addressed.
E-mail: herq@sun5.ibp.ac.cn;

Abstract

Recent studies have shown that neurodegeneration is closely
related to misfolding and aggregation of neuronal tau.

Our previous results show that neuronal tau aggregates in
formaldehyde solution and that aggregated tau induces apoptosis
of SH-SY5Y and hippocampal cells.

In the present study, based on atomic force microscopy (AFM)
observation, we have found that formaldehyde at low concentrations
induces tau polymerization whilst acetaldehyde does not.

Neuronal tau misfolds and aggregates into globular-like polymers
in 0.01 - 0.1% formaldehyde solutions.

Apart from globular-like aggregation, no fibril-like polymerization
was observed when the protein was incubated with formaldehyde
for 15 days.

SDS-PAGE results also exhibit tau polymerizing in the presence
of formaldehyde.

Under the same experimental conditions, polymerization of bovine
serum albumin (BSA) or a-synuclein was not markedly detected.

Kinetic study shows that tau significantly misfolds and polymerizes
in 60 minutes in 0.1% formaldehyde solution.

However, presence of 10% methanol prevents protein tau from
polymerization.

This suggests that formaldehyde polymerization is involved in tau
aggregation.

Such aggregation process is probably linked to the tau's special
"worm-like" structure, which leaves the e-amino groups of Lys
and thiol groups of Cys exposed to the exterior.

Such a structure can easily bond to formaldehyde molecules
in vitro and in vivo.

Polymerizing of formaldehyde itself results in aggregation of
protein tau.

Immunocytochemistry and thioflavin S staining of both endogenous
and exogenous tau in the presence of formaldehyde at low
concentrations in the cell culture have shown that formaldehyde can
induce tau into amyloid-like aggregates in vivo during apoptosis.

The significant protein tau aggregation induced by formaldehyde
and the severe toxicity of the aggregated tau to neural cells may
suggest that toxicity of methanol and formaldehyde ingestion is
related to tau misfolding and aggregation.

Funding: This project was supported by NSFB (06J11),
the NSFC (Nos. 90206041, 30570536 and 30621004)
and 973-Project (2006CB500703 and 2006CB911003).

Competing interests: The authors have declared that no competing
interests exist.

Academic Editor: Christophe Herman, Baylor College of Medicine,
United States of America

Introduction

Neuronal tau is an important protein in promoting and stabilizing the
microtubule system involved in cellular transport
and neuronal morphogenesis.

The tau molecule can be subdivided into an amino-terminal domain
that projects from the microtubule surface and a carboxy-terminal
microtubule-binding domain.

The discovery that incubation of bacterially expressed human tau
with sulphated glycosaminoglycans leads to bulk assembly of tau
filaments [1], making it possible to obtain structural information [2].

By using circular dichroism measurement, Schweer et al. have found
that protein tau lacks secondary structures and is considered in a
"worm-like" conformation with a high flexibility [3].

Therefore, the side-chains of amino acids such as Lys, Cys, Thr
and Ser are mostly exposed and vulnerable to chemical modification.

Recently, many laboratories have found that misfolding and
aggregation of protein tau are involved in neurodegeneration
[2], [4] - [6].

Protein tau has been found as the major component of paired
helical filaments in neurofibrillary tangles in the brains of Alzheimer's
patients, where abnormal hyper-phosphorylation induces tau to
misfold and form the paired helical filaments,
depositing in the cytoplasm of neurons [7] - [10].

Recently, a great deal of evidence has demonstrated that oxidation
and glycation stresses are key causal factors of neuronal degenerative
diseases [11] - [13].

Both of them inevitably produce a variety of unsaturated carbonyls
as intermediates, like malondialdehyde and 4-hydroxynonenal,
which usually cause carbonyl-amino crosslinking and lead to
accumulation of irreversible changes (like lipofuscin) related to
various neurodegenerative diseases in particular [14] - [16].

Such carbonyl stress-related reactions (carbonylation) can form
unstable and reversible 1:1 amino-carbonyl (Shiff's base)
compounds at an early stage of protein modification [16], [17].

Carbonylation binds and blocks a-/e- amino groups,
and results in changes in charge and conformation of a protein.

In order to investigate the relationship between carbonylation and
protein tau misfolding, the basic and simplest carbonyl compound
formaldehyde [18] has come into our attention.

Formaldehyde is a common environmental agent found in paint, cloth,
exhaust gas and many other medicinal and industrial products [19].

Formaldehyde exposure leads to formation of DNA/protein
crosslinks, a major mechanism of DNA damage.

The DNA/protein crosslinks have been used as a measure of dose
in drug delivery [20].

Formaldehyde, as a crosslinking agent, also reacts with thiol and
amino groups, leading to protein polymerization [21], [22].

Furthermore, methanol ingestion is an important public health
concern because of the selective actions of its toxic metabolites,
formaldehyde and formic acid, on the retina, the optic nerves
and the central nervous system (CNS) [23].

Illicit consumption of industrial methylated spirits can cause severe
and even fatal illness [24].

In the liver and retina, methanol is oxidized by alcohol
dehydrogenase, resulting in formaldehyde.

In semicarbazide-sensitive amine oxidase (SSAO)-mediated
pathogenesis of Alzheimer's disease, formaldehyde interacts
with B-amyloids and produces irreversibly cross-linked neurotoxic
amyloid-like complexes [21], [22], [25].

We have examined the role of formaldehyde
in misfolding of protein tau [26].

In particular, we investigated the toxicity of formaldehyde-induced
tau aggregates on human neuroblastoma cells (SH-SY5Y cell line)
and rat hippocampal cells [27].

The results showed that low concentrations (0.01 - 0.1%) of
formaldehyde are sufficient to induce formation of amyloid-like tau
aggregates, which can induce apoptosis of both SH-SY5Y
and hippocampal cells.

This may be significant to understand the mechanism of chronic
damage caused by methanol toxicity
and formaldehyde stress [18], [28].

However, we have still not known the mechanism of protein tau
aggregation in the presence of formaldehyde at low concentrations.

The present study concerns the characteristic of misfolding and
polymerization of extracellular and intracellular neuronal tau induced
by formaldehyde at low concentrations.....

Discussion

Clinical lethal dose of formaldehyde

Why did we investigate tau misfolding in the presence of
formaldehyde at low concentrations (0.01 - 0.1%)?

Methanol and ethanol are metabolized to formaldehyde and
acetaldehyde respectively in our hepatocytes
and some neural cells [36], [37].

Both formaldehyde and acetaldehyde can go through the
blood-brain barrier and cause some lesions to CNS,
especially our visual system [38].

Clinically, the lethal dose of formaldehyde for human beings is
about 0.08% in the circulation [39].

We have shown in the present study that formaldehyde can
significantly induce tau aggregation and polymerization at
concentrations even lower than 0.08%,
the clinical dose of toxicosis.

The same low concentration of formaldehyde did not induce
polymerization of BSA though theoretically it will cause any
protein to polymerize if the concentration is high enough.

On the other hand, although it is known that acetaldehyde is
acutely toxic and would covalently bind to proteins and other
macromolecules [40], in our AFM and SDS-PAGE studies
we did not observe tau polymerization caused by acetaldehyde at
the concentration range that we studied (0.1 - 1%)......

Tau aggregation relating to methanol and formaldehyde toxicity

Methanol is an ocular toxicant, which causes visual dysfunction and
often leads to blindness after acute exposure.

However, physiological and biochemical changes responsible
for the toxicity have not yet been well understood [28].

According to a recent report, humans are uniquely sensitive to the
toxicity of methanol, as they have limited capacity to oxidize and
detoxify formic acid.

Thus, the toxicity of methanol in humans is characterized by formic
acidaemia, metabolic acidosis, blindness or serious visual impairment,
mild central nervous system depression
and even death [23], [27], [28].

However, methanol toxicosis induces progressive complications
to CNS.

It is hard to explain the progressively chronic damage by local
accumulation of formic acid alone.

Therefore, the potential effect of formaldehyde on protein
misfolding may be significant, although formaldehyde remains
in the human body for only a short time.

In semicarbazide-sensitive amine oxidase (SSAO)-mediate
pathogenesis of Alzheimer's disease, formaldehyde interacts with
B-amyloids and produces irreversibly cross-linked neurotoxic
amyloid-like complexes [21], [22], [25].

Our studies showed that formaldehyde induced neuronal tau
to aggregate.

The amyloid-like tau induces apoptosis of SY5Y
and hippocampal cells [27].

In fact, chemically, formaldehyde reacts with thiol and
amino groups instantly,
resulting in subsequent misfolding of neuronal tau (Figure 11).

This suggests that amyloid-like tau is involved in methanol toxicosis,
especially the damage of neurons and the resulted complications
after exposure to formaldehyde.

Although there have been many studies on methanol and
formaldehyde intoxication [23], [24], none of them has addressed
the contribution of protein misfolding to the pathological mechanism,
in particular the effect of formaldehyde on protein conformation
and polymerization.

Interestingly, neurofibrillary tangles have been found in brains of
chronic alcoholics possessing neuropathological signs
of thiamine-deficiency [40], [47].

This suggests that tau misfolding may be involved in the
alcohol-induced pathological pathway.

Khlistunova and his colleagues found that neuronal tau repeat domain
could aggregate in vivo and was toxic to neuronal cells.

The degree of tau aggregation and toxicity depends on the propensity
of the B-structure [2], [48].

In the present study, we have demonstrated that amyloid-like
intracellular tau aggregates could induce cell apoptosis, a similar result
as that obtained for extracellular amyloid or a-synuclein [49] -- [51].

This suggests that an enriched B-sheet structure is important to
amyloid-like protein aggregation and neurotoxicity.

In our experiments, a low concentration of formaldehyde induced
both extracellular and intracellular tau proteins to aggregate into
cell-toxic amyloid-like granular aggregates [27].

It appears to provide a new mechanism for triggers of tauopathies
in the formaldehyde toxicosis.....

Acknowledgments

We thank Ms. Ya-Qun Zhang for technical assistance
and Dr. Ya-Jie Xu for providing the clone of HA-tau40.

Author Contributions

Conceived and designed the experiments: RH.
Performed the experiments: CN YW YL WD.
Analyzed the data: CN.
Wrote the paper: CN RH YL XC MD ST.

References.....

#19 Quievryn G, Zhitkovich A. (2000)
Loss of DNA-protein crosslinks from formaldehyde-exposed cells
occurs through spontaneous hydrolysis and an active repair process
linked to proteosome function.
Carcinogenesis 21: 1573 - 1580.

#20 Heck H, Casanova M. (1999)
Pharmacodynamics of formaldehyde: applications of a model for the
arrest of DNA replication by DNA-protein cross-links.
Toxicol Appl Pharmacol 160: 86 - 100.

#21 Yu PH, Lu LX, Fan H, Kazachkov M, Jiang ZJ, et al. (2006)
Involvement of semicarbazide-sensitive amine oxidase-mediated
deamination in lipopolysaccharide-induced
pulmonary inflammation.
Am J Pathol 168: 718 - 726.

#22 Yu PH. (2001)
Involvement of cerebrovascular semicarbazide-sensitive amine
oxidase in the pathogenesis of Alzheimer's disease
and vascular dementia.
Med Hypotheses 57: 175 - 179.

#23 Eells JT, Henry MM, Lewandowski MF, Seme MT,
Murray TG. (2000)
Development and characterization of a rodent model of
methanol-induced retinal and optic nerve toxicity.
Neurotoxicology 21: 321 - 330.

#24 Dayan AD, Paine AJ. (2001)
Mechanisms of chromium toxicity, carcinogenicity
and allergenicity: review of the literature from 1985 to 2000.
Hum Exp Toxicol 20: 439 - 451.

#25 Gubisne-Haberle D, Hill W, Kazachkov M,
Richardson JS, Yu PH. (2004)
Protein cross-linkage induced by formaldehyde derived from
semicarbazide-sensitive amine oxidase-mediated deamination
of methylamine.
J Pharmacol Exp Ther 310: 1125 - 1132.

#26 Nie CL, Zhang W, Zhang D, He RQ. (2005)
Changes in conformation of human neuronal tau during
denaturation in formaldehyde solution.
Protein Pept Lett 12: 75 - 78.

#27 Nie CL, Wang XS, Liu Y, Perrett S, He RQ. (2007)
Amyloid-like aggregates of neuronal tau induced by formaldehyde
promote apoptosis of neuronal cells.
BMC Neurosci 8: 9.

#28 Garner CD, Lee EW, Louis-Ferdinand RT. (1995)
Muller cell involvement in methanol-induced retinal toxicity.
Toxicol Appl Pharmacol 130: 101 - 107.

#32 Pomerantz M, Bittner S, Khader SB. (1982)
"Formaldehyde semicarbazone."
J Org Chem 47: 2217 - 2218.

#36 Barceloux DG, Bond GR, Krenzelok EP,
Cooper H, Vale JA. (2002)
American Academy of Clinical Toxicology practice guidelines
on the treatment of methanol poisoning.
J Toxicol Clin Toxicol 40: 415 - 446.

#37 Valentine WM. (1990)
Toxicology of selected pesticides, drugs, and chemicals.
Short-chain alcohols.
Vet. Clin. North Am. Small Anim. Pract 20: 515 - 523.

#38 Shcherbakova LN, Tel'pukhov VI, Trenin SO,
Bashilov IA, Lapkina TI. (1986)
[Permeability of the blood-brain barrier
to intra-arterial formaldehyde].
Biull Eksp Biol Med 102: 573 - 575.

[ Biull Eksp Biol Med. 1986 Nov; 102(11): 573-5.
[Permeability of the blood-brain barrier to intra-arterial
formaldehyde]
[Article in Russian]
Shcherbakova LN, Tel'pukhov VI, Trenin SO,
Bashilov IA, Lapkina TI.

Formaldehyde concentration was assessed in the brain,
cerebrospinal liquor, arterial and venous blood of intact animals
and following its intraarterial injections.

It is concluded that formaldehyde is capable of penetrating
through the blood-brain barrier, with the degree of permeability
depending on blood formaldehyde concentration.

The distribution of formaldehyde in the blood-brain-cerebrospinal
liquor system suggests the presence of both protein-bound
and unbound formaldehyde forms in the organism.
PMID: 3779084 ]

#39 Erkrath KD, Adebahr G, Kloppel A. (1981)
[Lethal intoxication by formalin during dialysis (author's transl)].
Z Rechtsmed 87: 233 - 236.

#40 Niemela O. (1999)
Aldehyde-protein adducts in the liver as a result of
ethanol-induced oxidative stress.
Front Biosci 4: D506 - D513.

#45 Jiang W, Schwendeman SP. (2000)
Formaldehyde-mediated aggregation of protein antigens:
comparison of untreated and formalinized model antigens.
Biotechnol Bioeng 70: 507 - 517.

#46 Rait VK, O'Leary TJ, Mason JT. (2004)
Modeling formalin fixation and antigen retrieval with
bovine pancreatic ribonuclease A:
I-structural and functional alterations.
Lab Invest 84: 292 - 299.

#47 Cullen KM, Halliday GM. (1995)
Neurofibrillary tangles in chronic alcoholics.
Neuropathol Appl Neurobiol 21: 312 - 318.
____________________________________________________



Note: many recent aspartame bans.....

http://groups.yahoo.com/group/aspartameNM/message/1426
ASDA (unit of Wal-Mart Stores WMT.N) and Marks & Spencer
will join Tesco and also Sainsbury to ban and limit aspartame,
MSG, artificial flavors dyes preservatives additives, trans fats, salt
"nasties" to protect kids from ADHD: leading UK media:
Murray 2007.05.15

http://groups.yahoo.com/group/aspartameNMmessage/1451
Artificial sweeteners (aspartame, sucralose) and coloring agents
will be banned from use in newly-born and baby foods,
the European Parliament decided: Latvia ban in schools 2006:
Murray 2007.07.12

http://groups.yahoo.com/group/aspartameNM/message/1341
Connecticut bans artificial sweeteners in schools, Nancy Barnes,
New Milford Times: Murray 2006.05.25

http://groups.yahoo.com/group/aspartameNM/message/1369
Bristol, Connecticut, schools join state program to limit artificial
sweeteners, sugar, fats for 8800 students, Johnny J Burnham,
The Bristol Press: Murray 2006.09.22


http://groups.yahoo.com/group/aspartameNM/message/1513
metabolic syndrome is tied to diet soda, PL Lutsey, LM Steffen,
J Stevens, Circulation 2008.01.22: role of formaldehyde and
formic acid from methanol in wines, liquors, or aspartame?:
Murray 2008.02.21

"But the one-third who ate the most fried food increased their risk
by 25 percent, compared with the one-third who ate the least, and
surprisingly, the risk of developing metabolic syndrome was 34
percent higher among those who drank one can of diet soda a day
compared with those who drank none.

"This is interesting," said Lyn M. Steffen, an associate professor of
epidemiology at the University of Minnesota and a co-author of the
paper, which was posted online in the journal Circulation on Jan. 22.
"Why is it happening? Is it some kind of chemical in the diet soda,
or something about the behavior of diet soda drinkers?""

"The diet soda association was not hypothesized
and deserves further study."

http://groups.yahoo.com/group/aspartameNM/message/1143
methanol (formaldehyde, formic acid) disposition:
Bouchard M et al, full plain text, 2001:
substantial sources are degradation
of fruit pectins, liquors, aspartame, smoke:
Murray 2005.04.02

http://groups.yahoo.com/group/aspartameNM/message/1511
vinyl acetate, ethyl alcohol, or aspartame in womb increases later
cancers in adults with lifetime exposure in many studies, M Soffritti
et al, Ramazzini Foundation, Basic Clin. Pharm. Toxicol. 2008 Feb.:
Rich Murray 2008.02.07

http://groups.yahoo.com/group/aspartameNM/message/1016
President Bush & formaldehyde (aspartame) toxicity:
Ramazzini Foundation carcinogenicity results Dec 2002:
Soffritti: Murray 2003.08.03 rmforall

p. 88 "The sweetening agent aspartame hydrolyzes in the
gastrointestinal tract to become free methyl alcohol,
which is metabolized in the liver
to formaldehyde, formic acid, and CO2. (11)"
Medinsky MA & Dorman DC. 1994;
Assessing risks of low-level methanol exposure.
CIIT Act. 14: 1-7.

http://groups.yahoo.com/group/aspartameNM/message/1453
Souring on fake sugar (aspartame), Jennifer Couzin,
Science 2007.07.06: 4 page letter to FDA from 12 eminent
USA toxicologists re two Ramazzini Foundation cancer studies
2007.06.25: Murray 2007.07.18

30 female pet store rats drinking lifelong 13.5 mg aspartame,
1/3 packet of Equal, had 33% with obvious tumors -- also bulging,
sick, and missing eyes, paralysis, obesity, skin sores -- agrees with
Ramazzini Foundation results, Victoria Inness-Brown:
Murray 2008.02.15
http://rmforall.blogspot.com/2008_02_01_archive.htm
Friday, February 15, 2008
http://groups.yahoo.com/group/aspartameNM/message/1521


http://groups.yahoo.com/group/aspartameNM/message/1490
details on 6 epidemiological studies since 2004 on diet soda (mainly
aspartame) correlations, as well as 14 other mainstream studies
on aspartame toxicity since summer 2005: Murray 2007.11.27

http://groups.yahoo.com/group/aspartameNM/message/1340
aspartame groups and books:
updated research review of 2004.07.16: Murray 2006.05.11

http://groups.yahoo.com/group/aspartameNM/message/1469
highly toxic formaldehyde, the cause of alcohol hangovers, is
made by the body from 100 mg doses of methanol from
dark wines and liquors, dimethyl dicarbonate, and aspartame:
Murray 2007.08.31


old tiger roars -- Woodrow C Monte, PhD -- aspartame causes
many breast cancers, as ADH enzyme in breasts makes methanol
from diet soda into carcinogenic formaldehyde -- same in dark
wines and liquors, Fitness Life 2008 Jan.: Murray 2008.02.11
http://rmforall.blogspot.com/2008_02_01_archive.htm
Monday, February 11, 2008
http://groups.yahoo.com/group/aspartameNM/message/1517

"Alcohol dehydrogenase ADH is required for the conversion of
methanol to formaldehyde (112).

ADH is not a common enzyme in the human body -- not many cells
in the human body contain this enzyme.

The human breast is one of the few organs in the body with a high
concentration of ADH (190b), and it is found there exclusively in the
mammary epithelial cells, the very cells known to transform into
adenocarcinoma (190c) (breast cancer).

The most recent breast cancer scientific literature implicates ADH
as perhaps having a pivotal role in the formation of breast cancer,
indicating a greater incidence of the disease in those
with higher levels of ADH activity in their breasts (190a)."

role of formaldehyde, made by body from methanol from foods
and aspartame, in steep increases in fetal alcohol syndrome, autism,
multiple sclerosis, lupus, teen suicide, breast cancer, Nutrition
Prof. Woodrow C. Monte, retired, Arizona State U., two reviews,
190 references supplied, Fitness Life, New Zealand
2007 Nov, Dec: Murray 2007.12.26
http://rmforall.blogspot.com/2007_12_01_archive.htm
Wednesday, December 26 2007
http://groups.yahoo.com/group/aspartameNM/message/1498


bias, omissions, incuriosity = opportunity, aspartame safety
evaluation, Magnuson BA, Burdock GA, Williams GM, 7 more,
2007 Sept, Ajinomoto funded 98 pages html [ $ 32 pdf ]:
Murray 2007.09.15
http://rmforall.blogspot.com/2007_09_01_archive.htm
Saturday, September 15, 2007

MSG and Aspartame -- A Personal Story, TV health reporter
Dick Allgire (vegetarian) healed of migraines and panic attacks:
Murray 2008.02.12
http://rmforall.blogspot.com/2008_02_01_archive.htm
Tuesday, February 12, 2008
http://groups.yahoo.com/group/aspartameNM/message/1520


"Of course, everyone chooses, as a natural priority, to enjoy
peace, joy, and love by helping to find, quickly share, and positively
act upon evidence about healthy and safe food, drink, and
environment."

Rich Murray, MA Room For All rmforall@comcast.net
505-501-2298 1943 Otowi Road, Santa Fe, New Mexico 87505

http://RMForAll.blogspot.com new primary archive

http://groups.yahoo.com/group/aspartameNM/messages
group with 120 members, 1,524 posts in a public archive

http://groups.yahoo.com/group/aspartame/messages
group with 1,077 members, 22,286 posts in public archive
____________________________________________________

Hawaii House Concurrent Resolution #132 for Health Department panel to decide aspartame ban by early 2010, Hawaii Rep. Josh Green MD, Health Committee
Chair: Murray 2008.03.12
http://rmforall.blogspot.com/2008_03_01_archive.htm
Wednesday, March 12, 2008
http://groups.yahoo.com/group/aspartameNM/message/1525
_____________________________________________________


www.capitol.hawaii.gov/session2008/bills/HCR132_.pdf

Hawaii House of Representatives H.C.R. No. 132,
Twenty-fourth Legislature,

State of Hawaii -- House Concurrent Resolution

Requesting the Director of Health to form a work group to explore
the need to ban or better label products containing the artificial
sweetener aspartame.

WHEREAS, discovered in 1965, aspartame was first approved in the
United States in 1981 and is one of the most widely used artificial
sweeteners; and

WHEREAS, when metabolized by the body, aspartame is broken
down into two common amino acids, aspartic acid and phenylalanine,
and a small amount of a third substance, methanol; and
[ Note: the aspartame molecule becomes equal numbers of aspartic
acid, phenylalanine, and methanol molecules (which although light weight,
are more chemically active by far, being turned quickly into nearly the
same number of molecules of formaldehyde and then formic acid,
enough to have toxic effects in every cell in the body by binding with
and impairing DNA, RNA, and protein molecules. ]

WHEREAS, these three substances occur in similar or greater amounts
in common foods; and
[ Note: methanol and formaldehyde are also supplied by dark wines
and liquors (the major cause of "morning after" hangovers), tobacco and
wood smoke, fumes from faulty furnaces, heaters and stoves, many fruits
and vegetables (when heated in canning, fermented, or degraded by
colonic bacteria), mobile homes and RVs, particleboard, new buildings
new cars, carpets, drapes, and furniture, mortuaries, medical training
and research, many cleaners and disinfectants, many personal care
products ranging from leather to hair dyes, and more, including many
Asian foods. ]

WHEREAS, aspartame continues to be the subject of strong public
controversy regarding its safety; and

WHEREAS, some studies have recommended further investigation into
the possible connection between aspartame and diseases such as brain
tumors, brain lesions, and lymphoma; and
[ Note: the range of symptoms also includes headache, fatigue, poor
memory, irritability, lassitude, mania, anxiety, depression, "brain fog",
insomnia, dizziness, many body pains and muscle cramps, restless legs,
interstitial cystitis, eye and vision problems, tinnitus, dry eyes and
mouth,
skin rashes, diarrhea or constipation, partial paralysis (often confused as
MS), ideopathic seizures, weight gain or loss, blood sugar swings,
craving and addiction, impaired fertility and pregnancy, "spontaneous"
abortion, premature birth, low birth weight, birth defects, sudden infant
death syndrome, autism, and early diabetes -- this dazzling litany of
outcomes is the natural result of chronic, long-term, low-level exposure
to formaldehyde and formic acid. Naturally, there are countless
interactions with other drugs and toxins, as well as major and complex
individual genetic variations.

Cheap, safe folic acid protects against formaldehyde toxicity in most
people. ]

WHEREAS, some human and animal studies have found adverse effects
and some have found no adverse effects; and [ Note: almost all industry
and vested interest funded research claims no evidence of problems,
while the opposite is found by independent teams. ]

WHEREAS, while there have been a number of studies on the effects
of aspartame on people, there is a great deal of argument by both
proponents and opponents on the methods used in the testing and
whether accurate representations of the consequences of average
consumption by people have been achieved by these studies; and
Note: a major need is to focus on the effects for long-term heavy users,
above 6 12-oz cans daily diet drink, from vulnerable groups like fetus,
infants, kids, teens, students, difficult critical jobs like police, pilots,
military, and nuclear plant operators, pregnant women, seniors, obese,
diabetic, depressed, violent criminal, cancer, and, especially,
over-worked PR agents lavishly funded to defend aspartame as,
"the most tested food additive in history".

WHEREAS, the Internet has become a tool for many to spread and
promote the opinion they adhere to as fact whether it is that aspartame
is a neurotoxin derived from toxic sludge or it is a harmless product
enabling a healthy lifestyle; and

WHEREAS, while many may find that the simple answer to the
problem is to not purchase or use any product containing aspartame
in it, there are those who maintain that some products do not
specifically list aspartame as an ingredient, rather it is hidden under
the label of "natural flavors"; now, therefore,

BE IT RESOLVED by the House of Representatives of the
Twenty-fourth Legislature of the State of Hawaii, Regular Session
of 2008, the Senate concurring, that the Director of Health is
requested to form a work group to explore the need to ban or
improve labeling for products containing the artificial sweetener
aspartame; and

BE IT FURTHER RESOLVED the members of the work group
should include:

(1) One member from the House of Representatives
appointed by the Speaker of the House of Representatives;

(2) One member of the Senate
appointed by the President of the Senate;

(3) The Director of Health or the director's designee;

(4) The President of the Calorie Control Council or the President's
designee; and [ the preminent aspartame lobby ]

(5) The President and Chief Executive Officer
of the American Beverage Association or a designee;
[ With their original name, The National Soft Drink Association, they
condemned aspartame on July 28, 1983 in a detailed critique:
http://dorway.com/dorwblog/?page_id=60 ]

(6) The President of the Hawaii Society of Naturopathic Physicians or a
designee;

(7) The Founder of Mission Possible International or a designee; and

(8) Two consumers appointed by the Director of Health; and

BE IT FURTHER RESOLVED the work group is requested to submit
a report of its findings, as well as any suggested legislation, to the
Legislature no later than 20 days prior to the convening of the
Regular Session of 2010; and

BE IT FURTHER RESOLVED that certified copies of this
Concurrent Resolution be transmitted to the Director of Health,
President of the Calorie Control Council,
webmaster@caloriecontrol.org;
President and Chief Executive Officer
of the American Beverage Association, info@ameribev.org;
President of the Hawaii Society of Naturopathic Physicians,
dr_nguyen@msn.com;
and Founder of Mission Possible International, Betty Martini,
bettm19@mindspring.com


Offered by Josh Green, MD March 4, 2008

Honorable Rep. Josh Green, M.D., Chair, Health Committee
repgreen@Capitol.hawaii.gov;
6th Representative District
Hawaii State Capitol, Room 327
415 South Beretania Street
Honolulu, HI 96813
phone 808-586-9605; fax 808-586-9608
From the Big Island, toll free 974-4000 + 69605

Hawaiian aspartame ban bills in House and Senate challenge
corporate clout, Sen. J. Kalani English & Suzanne Chun Oakland,
Rep. Calvin K.Y. Say & Mele Carroll: Murray 2008.01.25
http://rmforall.blogspot.com/2008_01_01_archive.htm
Friday, January 25, 2008
http://groups.yahoo.com/group/aspartameNM/message/1505
_____________________________________________________

www.hawaiind.org/index.htm
Hawaii Society of Naturopathic Physicians
750-D Kapahulu Ave
Honolulu, HI 96816 808-732-6996
Dr. Ye Nguyen - President dr_nguyen@msn.com;
Dr. Monique Yuen - Vice President: drmoniqueyuen@gmail.com;
Dr. Karen Tan - Treasurer
Dr. Madeleine Portuondo madeleine_portuondo@yahoo.com;
_____________________________________________________



methanol impurity in alcohol drinks [ and aspartame ] is turned into
neurotoxic formic acid, prevented by folic acid, re Fetal Alcohol
Syndrome, BM Kapur, DC Lehotay, PL Carlen at U. Toronto,
Alc Clin Exp Res 2007 Dec. plain text: detailed biochemistry,
CL Nie et al. 2007.07.18: Rich Murray 2008.02.24
http://rmforall.blogspot.com/2008_02_01_archive.htm
Sunday, February 24, 2008
http://groups.yahoo.com/group/aspartameNM/message/1524


[ Rich Murray comments: As a medical layman volunteer information
activist for aspartame and related toxicity issues since January 1999,
I note with appreciation the remarkable exponential progress on all
fronts, including a rapidly emerging consensus about the primary
importance of all toxicity challenges for our world.

This lengthy review features in detail two quite different, revolutionary
contributions, from Canada, and England and China.

It is indicative of our times that the CL Nie et al. study, 2007
appears in a free, open access journal-- indeed,
as all life and death information must.

Following rather vigorously, indeed blindly, the imperatives of
single-minded, profit-driven capitalist competition -- manipulating
adroitly research, education, media, citizens, governments -- many
great global corporations have inevitably created results that
oppose the common good. Alcohol and tobacco are well known.

Realistically, any further manipulations can only lead to inevitable
and even sudden corporate meltdowns, in the context of an
unfettered, cooperative, democratic global information forum,
the Internet.

Now, it is as easy and cheap to compose and instantly post a
30-page review as 3 pages a decade ago -- and such reviews
are archived forever in multiple collections, open via global search
engines to a billion Net citizens.

Perforce, and increasingly happily, all societal entities will have to
operate by high and shared voluntary universal standards
for the common good. ]


http://www.blackwell-synergy.com/doi/abs/10.1111/j.1530-0277.2007.00541.x

Alcoholism: Clinical and Experimental Research
Volume 31 Issue 12 Page 2114-2120, December 2007

Bhushan M. Kapur, b.kapur@utoronto.ca;
Arthur C. Vandenbroucke, PhD, FCACB
Yana Adamchik,
Denis C. Lehotay, dlehotay@health.gov.sk.ca;
Peter L. Carlen carlen@uhnres.utoronto.ca;
(2007) Formic Acid, a Novel Metabolite of Chronic Ethanol
Abuse, Causes Neurotoxicity, Which Is Prevented by Folic Acid
Alcoholism: Clinical and Experimental Research 31 (12), 2114-2120.
doi:10.1111/j.1530-0277.2007.00541.x

Abstract

Background:
Methanol is endogenously formed in the brain and is present as a
congener in most alcoholic beverages.

Because ethanol is preferentially metabolized over methanol (MeOH)
by alcohol dehydrogenase, it is not surprising that MeOH
accumulates in the alcohol-abusing population.

This suggests that the alcohol-drinking population will have higher
levels of MeOH's neurotoxic metabolite, formic acid (FA).

FA elimination is mediated by folic acid.

Neurotoxicity is a common result of chronic alcoholism.

This study shows for the first time that FA,
found in chronic alcoholics, is neurotoxic
and this toxicity can be mitigated by folic acid administration.

Objective:
To determine if FA levels are higher in the alcohol-drinking
population and to assess its neurotoxicity in organotypic
hippocampal rat brain slice cultures.

Methods:
Serum and CSF FA was measured in samples from both ethanol
abusing and control patients, who presented to a hospital emergency
department. [ CSF = Cerebral Spinal Fluid ]

FA's neurotoxicity and its reversibility by folic acid were assessed
using organotypic rat brain hippocampal slice cultures using clinically
relevant concentrations.

Results:
Serum FA levels in the alcoholics
(mean ± SE: 0.416 +- 0.093 mmol/l, n = 23)
were significantly higher than in controls
(mean ± SE: 0.154 +- 0.009 mmol/l, n = 82) (p < 0.0002).

FA was not detected in the controls' CSF (n = 20),
whereas it was >0.15 mmol/l in CSF of 3 of the 4 alcoholic cases.

Low doses of FA from 1 to 5 mmol/l added for 24, 48 or 72 hours
to the rat brain slice cultures caused neuronal death as measured by
propidium iodide staining.

When folic acid (1 umol/l) was added with the FA,
neuronal death was prevented. [ umol = micromole ]

Conclusions:
Formic acid may be a significant factor in the neurotoxicity of
ethanol abuse.

This neurotoxicity can be mitigated by folic acid administration
at a clinically relevant dose.

Key Words:
Formic Acid, Folic Acid, Methanol, Neurotoxicity, Alcoholism.
____________________________________________________


Note: many recent aspartame bans.....

http://groups.yahoo.com/group/aspartameNM/message/1426
ASDA (unit of Wal-Mart Stores WMT.N) and Marks & Spencer
will join Tesco and also Sainsbury to ban and limit aspartame,
MSG, artificial flavors dyes preservatives additives, trans fats, salt
"nasties" to protect kids from ADHD: leading UK media:
Murray 2007.05.15

http://groups.yahoo.com/group/aspartameNMmessage/1451
Artificial sweeteners (aspartame, sucralose) and coloring agents
will be banned from use in newly-born and baby foods,
the European Parliament decided: Latvia ban in schools 2006:
Murray 2007.07.12

http://groups.yahoo.com/group/aspartameNM/message/1341
Connecticut bans artificial sweeteners in schools, Nancy Barnes,
New Milford Times: Murray 2006.05.25

http://groups.yahoo.com/group/aspartameNM/message/1369
Bristol, Connecticut, schools join state program to limit artificial
sweeteners, sugar, fats for 8800 students, Johnny J Burnham,
The Bristol Press: Murray 2006.09.22


http://groups.yahoo.com/group/aspartameNM/message/1513
metabolic syndrome is tied to diet soda, PL Lutsey, LM Steffen,
J Stevens, Circulation 2008.01.22: role of formaldehyde and
formic acid from methanol in wines, liquors, or aspartame?:
Murray 2008.02.21

"But the one-third who ate the most fried food increased their risk
by 25 percent, compared with the one-third who ate the least, and
surprisingly, the risk of developing metabolic syndrome was 34
percent higher among those who drank one can of diet soda a day
compared with those who drank none.

"This is interesting," said Lyn M. Steffen, an associate professor of
epidemiology at the University of Minnesota and a co-author of the
paper, which was posted online in the journal Circulation on Jan. 22.
"Why is it happening? Is it some kind of chemical in the diet soda,
or something about the behavior of diet soda drinkers?""

"The diet soda association was not hypothesized
and deserves further study."

http://groups.yahoo.com/group/aspartameNM/message/1143
methanol (formaldehyde, formic acid) disposition:
Bouchard M et al, full plain text, 2001:
substantial sources are degradation
of fruit pectins, liquors, aspartame, smoke:
Murray 2005.04.02

http://groups.yahoo.com/group/aspartameNM/message/1511
vinyl acetate, ethyl alcohol, or aspartame in womb increases later
cancers in adults with lifetime exposure in many studies, M Soffritti
et al, Ramazzini Foundation, Basic Clin. Pharm. Toxicol. 2008 Feb.:
Rich Murray 2008.02.07

http://groups.yahoo.com/group/aspartameNM/message/1016
President Bush & formaldehyde (aspartame) toxicity:
Ramazzini Foundation carcinogenicity results Dec 2002:
Soffritti: Murray 2003.08.03 rmforall

p. 88 "The sweetening agent aspartame hydrolyzes in the
gastrointestinal tract to become free methyl alcohol,
which is metabolized in the liver
to formaldehyde, formic acid, and CO2. (11)"
Medinsky MA & Dorman DC. 1994;
Assessing risks of low-level methanol exposure.
CIIT Act. 14: 1-7.

http://groups.yahoo.com/group/aspartameNM/message/1453
Souring on fake sugar (aspartame), Jennifer Couzin,
Science 2007.07.06: 4 page letter to FDA from 12 eminent
USA toxicologists re two Ramazzini Foundation cancer studies
2007.06.25: Murray 2007.07.18

30 female pet store rats drinking lifelong 13.5 mg aspartame,
1/3 packet of Equal, had 33% with obvious tumors -- also bulging,
sick, and missing eyes, paralysis, obesity, skin sores -- agrees with
Ramazzini Foundation results, Victoria Inness-Brown:
Murray 2008.02.15
http://rmforall.blogspot.com/2008_02_01_archive.htm
Friday, February 15, 2008
http://groups.yahoo.com/group/aspartameNM/message/1521


http://groups.yahoo.com/group/aspartameNM/message/1490
details on 6 epidemiological studies since 2004 on diet soda (mainly
aspartame) correlations, as well as 14 other mainstream studies
on aspartame toxicity since summer 2005: Murray 2007.11.27

http://groups.yahoo.com/group/aspartameNM/message/1340
aspartame groups and books:
updated research review of 2004.07.16: Murray 2006.05.11

http://groups.yahoo.com/group/aspartameNM/message/1469
highly toxic formaldehyde, the cause of alcohol hangovers, is
made by the body from 100 mg doses of methanol from
dark wines and liquors, dimethyl dicarbonate, and aspartame:
Murray 2007.08.31


old tiger roars -- Woodrow C Monte, PhD -- aspartame causes
many breast cancers, as ADH enzyme in breasts makes methanol
from diet soda into carcinogenic formaldehyde -- same in dark
wines and liquors, Fitness Life 2008 Jan.: Murray 2008.02.11
http://rmforall.blogspot.com/2008_02_01_archive.htm
Monday, February 11, 2008
http://groups.yahoo.com/group/aspartameNM/message/1517

"Alcohol dehydrogenase ADH is required for the conversion of
methanol to formaldehyde (112).

ADH is not a common enzyme in the human body -- not many cells
in the human body contain this enzyme.

The human breast is one of the few organs in the body with a high
concentration of ADH (190b), and it is found there exclusively in the
mammary epithelial cells, the very cells known to transform into
adenocarcinoma (190c) (breast cancer).

The most recent breast cancer scientific literature implicates ADH
as perhaps having a pivotal role in the formation of breast cancer,
indicating a greater incidence of the disease in those
with higher levels of ADH activity in their breasts (190a)."

role of formaldehyde, made by body from methanol from foods
and aspartame, in steep increases in fetal alcohol syndrome, autism,
multiple sclerosis, lupus, teen suicide, breast cancer, Nutrition
Prof. Woodrow C. Monte, retired, Arizona State U., two reviews,
190 references supplied, Fitness Life, New Zealand
2007 Nov, Dec: Murray 2007.12.26
http://rmforall.blogspot.com/2007_12_01_archive.htm
Wednesday, December 26 2007
http://groups.yahoo.com/group/aspartameNM/message/1498


bias, omissions, incuriosity = opportunity, aspartame safety
evaluation, Magnuson BA, Burdock GA, Williams GM, 7 more,
2007 Sept, Ajinomoto funded 98 pages html [ $ 32 pdf ]:
Murray 2007.09.15
http://rmforall.blogspot.com/2007_09_01_archive.htm
Saturday, September 15, 2007

MSG and Aspartame -- A Personal Story, TV health reporter
Dick Allgire (vegetarian) healed of migraines and panic attacks:
Murray 2008.02.12
http://rmforall.blogspot.com/2008_02_01_archive.htm
Tuesday, February 12, 2008
http://groups.yahoo.com/group/aspartameNM/message/1520


"Of course, everyone chooses, as a natural priority, to enjoy
peace, joy, and love by helping to find, quickly share, and positively
act upon evidence about healthy and safe food, drink, and
environment."

Rich Murray, MA Room For All rmforall@comcast.net
505-501-2298 1943 Otowi Road, Santa Fe, New Mexico 87505

http://RMForAll.blogspot.com new primary archive

http://groups.yahoo.com/group/aspartameNM/messages
group with 120 members, 1,525 posts in a public archive

http://groups.yahoo.com/group/aspartame/messages
group with 1,080 members, 22,439 posts in public archive
_____________________________________________________