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 20097.09.01
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 β-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.plosone.org/article/fetchArticle.action?articleURI=info:doi/10.1371/j\
ournal.pone.0000629
free full text

Formaldehyde at Low Concentration Induces Protein Tau into Globular Amyloid-Like
Aggregates In Vitro and In Vivo
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@...,
Saul J.B. Tendler 2, Saul.Tendler@...,
Rong Giao He 1* herq@...,
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

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 α-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 ε-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

Citation: Nie CL, Wei Y, Chen X, Liu YY, Dui W, et al. (2007) Formaldehyde at
Low Concentration Induces Protein Tau into Globular Amyloid-Like Aggregates In
Vitro and In Vivo. PLoS ONE 2(7): e629. doi:10.1371/journal.pone.0000629

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@...

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 α-/ε- 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 attentions.

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 β-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 β-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
β-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 α-synuclein [49] -- [51].

This suggests that an enriched β-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.
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#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. Find this article online

#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.
Find this article online

#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. Find this article online

#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. Find this article online

#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. Find this article online

#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. Find this article online

#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. Find this article online

#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. Find this article online

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

#32 Pomerantz M, Bittner S, Khader SB. (1982) “Formaldehyde semicarbazone.”
J Org Chem 47: 2217 -- 2218. Find this article online

#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. Find this article online

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

#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.

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

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

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

#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. Find this article online

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

http://groups.yahoo.com/group/aspartameNM/message/1286
methanol products (formaldehyde and formic acid) are main cause of alcohol
hangover symptoms [same as from similar amounts of methanol, the 11% part of
aspartame]: YS Woo et al, 2005 Dec: Murray 2006.01.20

Addict Biol. 2005 Dec;10(4): 351-5.
Concentration changes of methanol in blood samples during
an experimentally induced alcohol hangover state.
Woo YS, Yoon SJ, Lee HK, Lee CU, Chae JH, Lee CT, Kim DJ.
Chuncheon National Hospital, Department of Psychiatry,
The Catholic University of Korea, Seoul, Korea.
http://www.cuk.ac.kr/eng/ sysop@...

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://www.toxsci.oupjournals.org/cgi/content/full/64/2/169


Fully 11 % of aspartame is methanol -- 1,120 mg aspartame
in 2 L diet soda, almost six 12-oz cans, gives 123 mg methanol
(wood alcohol). The methanol is immediately released
into the body after drinking .
Within hours, the liver turns much of the methanol into formaldehyde,
and then much of that into formic acid, both of which in time
are partially eliminated as carbon dioxide and water.

However, about 30 % of the methanol remains in the body
as cumulative durable toxic metabolites of formaldehyde
and formic acid -- 37 mg daily,
a gram every month, accumulating in and affecting every tissue.

If only 10 % of the methanol is retained daily as formaldehyde,
that would give 12 mg daily formaldehyde accumulation -- about
60 times more than the 0.2 mg from 10 % retention
of the 2 mg EPA daily limit for formaldehyde in drinking water.

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

http://groups.yahoo.com/group/aspartameNM/message/835
ATSDR: EPA limit 1 ppm formaldehyde in drinking water July 1999:
Murray 2002.05.30

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


http://groups.yahoo.com/group/aspartameNM/message/1464
13 mainstream research studies in 24 months showing aspartame
toxicity, also 3 relevant studies on methanol and formaldehyde: Murray
2007.08.17

http://groups.yahoo.com/group/aspartameNM/message/1468
Formaldehyde induced urticarial vasculitis in male medical student, age 40,
Michael Pellizzari, Gillian Marshman, Flinders U., Australasian J. Dermatol.
2007 Aug: Murray 2007.08.29

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


http://groups.yahoo.com/group/aspartameNM/message/1457
aspartame bans, tis more an avalanche than a trend...: Rich Murray
2007.08.17

So far, USA print and broadcast media are deaf, blind, and dumb,
regarding recent major bans of aspartame and MSG in the UK and EU.

The EU Parliament voted July 12 to ban artificial sweeteners
in newly born and infant foods.

On May 15 four huge UK supermarket chains announced bans
of aspartame and MSG, food dyes, and many additives
to protect kids from ADHD --
Sainsbury, Tesco, Marks & Spencer, and ASDA, a unit of WalMart.

May 31: Coca-Cola and the much larger Cargill Inc.,
after years of secret development, with 24 patents,
will soon sell rebiana (stevia) in drinks and food
in the many nations where it is approved as a sweetener --
for decades a major sweetener in Japan, China, Korea, Taiwan,
Thailand, Malasia, Saint Kitts, Nevis,
Brazil, Peru, Paraguay, Uruguay, and Israel,
and an approved supplement in USA, Australia, and Canada,
according to Wikipedia.

http://groups.yahoo.com/group/aspartameNM/message/1454
recent research and news re aspartame and stevia: Murray 2007.08.16

"Of course, everyone chooses, as a natural priority,
to actively find, quickly share, and positively act
upon the facts about healthy and safe food, drink,
and environment."

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

http://groups.yahoo.com/group/aspartameNM/messages
group with 82 members, 1,470 posts in a public,
searchable archive http://RMForAll.blogspot.com


http://groups.yahoo.com/group/aspartameNM/message/1395
Aspartame Controversy, in Wikipedia democratic
encyclopedia, 72 references (including AspartameNM # 864
and 1173 by Murray, brief fair summary of much more research:
Murray 2007.01.01

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/aspartameNMmessage/1437
stevia to be approved and cyclamates limited by
Food Standards Australia New Zealand:
JMC Geuns critiques of two recent stevia studies by Nunes:
Murray 2007.05.29

http://groups.yahoo.com/group/aspartameNM/message/1427
more from The Independent, UK, Martin Hickman, re ASDA
(unit of Wal-Mart Stores) and Marks & Spencer ban of
aspartame, MSG, artificial chemical additives and dyes
to prevent ADHD in kids: urray 2007.05.16
http://news.independent.co.uk/uk/health_medical/article2548747.ece

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/aspartameNM/message/1438
Coca-Cola and Cargill Inc., after years of development,
with 24 patents, will soon sell rebiana (stevia)
in drinks and foods: Murray 2007.05.31

www.ncbi.nlm.nih.gov/sites/entrez search PubMed
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