formaldehyde, aspartame, and migraines, the first case series, Sharon E Jacob-Soo, Sarah A Stechschulte, UCSD, Dermatitis 2008 May: Rich Murray 2008.07.18 http://rmforall.blogspot.com/2008_07_01_archive.htm
Friday, July 18, 2008 http://groups.yahoo.com/group/aspartameNM/message/1553
___________________________________________________

Dermatitis. 2008 May-Jun; 19(3): E10-1.
Formaldehyde, aspartame, and migraines: a possible connection.
Jacob SE, Stechschulte S.
Department of Dermatology and Cutaneous Surgery, University of Miami,
Miami, FL, USA.

Aspartame is a widely used artificial sweetener that has been linked to pediatric and adolescent migraines.

Upon ingestion, aspartame is broken, converted, and oxidized into formaldehyde in various tissues.

We present the first case series of aspartame-associated migraines related to clinically relevant positive reactions to formaldehyde on patch testing. PMID: 18627677

formaldehyde from many sources, including aspartame, is major cause of
Allergic Contact Dermatitis, SE Jacob, T Steele, G Rodriguez, Skin and
Aging 2005 Dec.: Murray 2008.03.27 http://rmforall.blogspot.com/2008_03_01_archive.htm
Thursday, March 27, 2008 http://groups.yahoo.com/group/aspartameNM/message/1533

"For example, diet soda and yogurt containing aspartame (Nutrasweet),
release formaldehyde in their natural biological degradation.

One of aspartame's metabolites, aspartic acid methyl ester, is
converted to methanol in the body, which is oxidized to formaldehyde
in all organs, including the liver and eyes. 22

Patients with a contact dermatitis to formaldehyde have been seen to
improve once aspartame is avoided. 22

Notably, the case that Hill and Belsito reported had a 6-month history
of eyelid dermatitis that subsided after 1 week of avoiding diet soda.
22"

Avoiding formaldehyde allergic reactions in children, aspartame,
vitamins, shampoo, conditioners, hair gel, baby wipes, Sharon E Jacob,
MD, Tace Steele, U. Miami, Pediatric Annals 2007 Jan.: eyelid contact
dermatitis, AM Hill, DV Belsito, 2003 Nov.: Murray 2008.03.27 http://rmforall.blogspot.com/2008_03_01_archive.htm
Thursday, March 27, 2008 http://groups.yahoo.com/group/aspartameNM/message/1532

Sharon E. Jacob, MD, Assistant Professor of Medicine (Dermatology)
University of California, San Diego 200 W. Arbor Drive #8420, San
Diego, CA 92103-8420
Tel: 858-552-8585 ×3504 Fax: 305-675-8317 sjacob@contactderm.net;

Dermatitis. 2008 Jan-Feb;19(1):9-15.
Systemic contact dermatitis.
Jacob SE, Zapolanski T. tamar.zapolanski@gmail.com;
Department of Dermatology and Cutaneous Surgery, University of Miami,
Miami, FL, USA.

Systemic exposure to allergens resulting in a cutaneous eruption is
known as systemic contact dermatitis (SCD).

Once sensitization occurs, varying exposures to antigens via multiple
routes (including transepidermal routes, intravenous or intramuscular
routes, inhalation, and ingestion) can result in systemic flare.

This article highlights the different categories of common
contactants, metals, medications, and plants, exposure to which leads
to SCD.

A comprehensive approach that takes into account all possible routes
of exposure is essential in diagnosing SCD and in helping patients
successfully avoid their allergens. PMID: 18346390

"We present a case of a medical student who presented with
erythematous eczematoid plaques on her trunk and legs and fine
vesiculation of her scalp, 3 weeks after starting anatomy class.

Of note, she routinely washed her face and arms after leaving the
anatomy lab, but remained in her scrubs for the rest of the day.

Formaldehyde and Quaternium-15 positive reactions in the same patient.
[ photo ]"

"Our patient underscores the importance of appropriate patch testing
and education.

Once we identified the allergy to formaldehyde and quaternium-15, we
provided patient education materials regarding the common and not-so-
common locations of these chemicals and cross-reactors.

We also gave the patient information on avoidance and safe
alternatives (see Table 5).

Fortunately, with technical advances, this student completed the
anatomy section via electronic learning tools.

By avoiding formaldehyde, including anatomy lab, FRP in her shampoo
and cosmetics, and aspartame in her diet, this patient dramatically
improved.

As with all contact dermatitides, the mainstay of treatment for
allergic contact dermatitis is avoidance."

http://www.skinandaging.com/article/5158 Skin & Aging Journal
Skin & Aging - ISSN: 1096-0120 - Volume 13 - Issue 12_2005 -
December 2005 - Pages: 22 - 27

Allergen Focus:
Focus on T.R.U.E. Test Allergens #21, 13 and 18:
Formaldehyde and Formaldehyde-Releasing Preservatives
– By Sharon E. Jacob, M.D., Tace Steele, B.A., [now MD] and Georgette
Rodriguez, M.D., M.P.H.

http://www.eczemacenter.org/eczema_center/meetfacultystaff.htm
[ photo ]

The Eczema Center
Rady Children's Hospital of San Diego
8010 Frost Street, Suite 602, San Diego, CA 92123
or call... (858) 966-6774

Sharon E. Jacob , MD
Dr. Sharon E. Jacob is Assistant Clinical Professor of Pediatrics and
Medicine (Dermatology) at the University of California, School of
Medicine and Rady Children's Hospital.
She earned her medical degree from the Temple University, and
completed dermatology training at the University of Miami and advanced
contact dermatitis training at New York University (NYU).
She has been board certified in dermatology.

Dr. Jacob's clinical interests include atopic and contact dermatitis
and education.
She is considered a national expert on chemical sensitivities in the
skin and has published more than 45 journal articles, book chapters
and abstracts on this topic.
In 2005, Dr Jacob was the first to present contact dermatitis data on
U.S. pediatric patients to the American Contact Dermatitis Society
(ACDS).

She has received an excellence in teaching award from the University
of Miami Dermatology and the Clinical Research Award from the ACDS.
She is an active reviewer for the following medical publications
including Journal of the American Academy of Dermatology, Pediatric
Dermatology, Dermatitis, and the Archives of Dermatology.
Dr. Jacob also serves on the medical board of the Inflammatory Skin
Disease Institute and the Skin and Aging Journal.

Dr. Jacob enjoys taking care of children and their families and is an
advocate for children's dermatologic health.

http://www.eczemacenter.org/eczema_center/index.htm

Atopic dermatitis (AD) – better known as eczema – is the most common
chronic skin disorder seen in infants and children.
In fact, the prevalence of this condition has risen dramatically
during the last three decades.
Currently, 15% to 20% of children in the United States are expected to
experience this condition sometime during their lifetime, compared to
7% around 1960.

The negative impact of eczema is profound and insidious.
It affects both the patient who suffers from it and that patient's
family members, and it does so on two important levels – physical and
emotional.

Physical:

Inflamed, itchy rashes can involve any and all of the skin surfaces
and are frequently complicated by skin breakdown and bacterial, viral,
and fungal infections.

It is linked to the development of life-long allergic conditions,
including asthma, food allergies, and rhinitis.

Any level of AD is extremely uncomfortable and, at times, painful.
Individuals with moderate to severe disease report that eczema hugely
disturbs their sleep and impacts performance of daily activities,
including adverse effects on school, sports activities, work, and peer
relationships.

In studies, individuals with eczema reported more negative impact on
quality of life than those with insulin-dependent diabetes!

Emotional:

Patients and their families experience considerable emotional
distress, anxiety, and embarrassment because of people's response to
this illness.

In fact, the emotional scarring on both patient and family members may
outlast eczema's physical effects.

Parents especially suffer because it is difficult for children
experiencing this condition to understand that their parents cannot
make the torment go away.
The stress of caring for these children is even greater than parents
caring for a child with insulin-dependent diabetes.

Patients experience considerable discrimination and social isolation
because of this illness.
People often stare, shiver with disgust or step back in fear from
those who have this condition.
The end result for patients: A life-time of struggle with their sense
of worth and self esteem.

http://aad2008.omnibooksonline.com/data/papers/CRS-113-F.pdf lecture
with photos
___________________________________________________

similar levels of daily formaldehyde and formic acid, causes of birth
defects, come from cigarettes, aspartame, and dark wines and liquors
– folic acid protects most people: Rich Murray 2008.07.15 http://rmforall.blogspot.com/2008_07_01_archive.htm
Tuesday, July 15, 2008 http://groups.yahoo.com/group/aspartameNM/message/1552

http://www.divine.ca/en/health-and-wellness/articles/c_16_i_3295/5-reasons-to-quit-smoking-1.html

"A smoker who goes through one pack a day will smoke 7,300 cigarettes
a year, inhaling the equivalent of nearly 1 gram of formaldehyde
(yikes!)."

That's about 2.5 mg daily formaldehyde intake for 20 cigarettes, over
the 2 mg USA FDA alarm level for formaldehyde in average 2 liters
daily drinking water, while a single 12 oz can of diet soda also
results in about 2 mg formaldehyde toxic products in the body,
including formic acid, a notorious cause of birth defects.

Dark wines and liquors usually supply even more methanol, which the
body always turns into formaldehyde and formic acid – the major cause
of "morning after" hangovers.

High levels of folic acid, a safe, affordable vitamin in fruits and
vegetables, largely prevents formaldehyde and formic acid toxicity in
most people.

It is certain that high levels of aspartame use, above 2 liters daily
for months and years, must lead to chronic formaldehyde-formic acid
toxicity.

Fully 11 % of aspartame is methanol – 1,120 mg aspartame in 2 liters
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 liters of
water.

formaldehyde and formic acid in FEMA trailers and other sources
(aspartame, dark wines and liquors, tobacco smoke): Murray 2008.01.30 http://rmforall.blogspot.com/2008_01_01_archive.htm
Wednesday, January 30, 2008 http://groups.yahoo.com/group/aspartameNM/message/1508

The FEMA trailers give about the same amount of formaldehyde and
formic acid daily as from a quart of dark wine or liquor, or two
quarts (6 12-oz cans) of aspartame diet soda, from their over 1 tenth
gram methanol impurity (one part in 10,000), which the body quickly
makes into formaldehyde and then formic acid – enough to be the major
cause of "morning after" alcohol hangovers.

Methanol and formaldehyde and formic acid also result from many fruits
and vegetables, tobacco and wood smoke, heater and vehicle exhaust,
household chemicals and cleaners, cosmetics, and new cars, drapes,
carpets, furniture, particleboard, mobile homes, buildings, leather...
so all these sources add up and interact with many other toxic
chemicals.

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: 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

opportunities re BA Magnuson, GA Burdock et al., Aspartame Safety
Evaluation 2007 Sept., Critical Reviews in Toxicology:
Rich Murray 2008.07.11 [ major review 67 pages ] http://rmforall.blogspot.com/2008_07_01_archive.htm
Friday, July 11, 2008
___________________________________________________

"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 126 members, 1,553 posts in a public archive

http://groups.yahoo.com/group/aspartame/messages
group with 1,125 members, 22,834 posts in public archive
___________________________________________________

folic acid prevents harm from formaldehyde and formic acid, formed from
methanol from many sources: Rich Murray 2008.07.10

re "A Few too Many", Joan Acocella, The New Yorker, long review of hangover
research 2008.05.26 – same levels of formaldehyde and formic acid in FEMA
trailers and other sources (aspartame, dark wines and liquors, tobacco
smoke): Murray 2008.06.05 http://rmforall.blogspot.com/2008_06_01_archive.htm
Thursday, June 5, 2008 http://groups.yahoo.com/group/aspartameNM/message/1541

[ See also:
There really is no controversy, Adrienne Samuels PhD, letter re
evident toxicity of aspartame EJCN 2008.06.11:
Murray 2008.06.30 http://rmforall.blogspot.com/2008_06_01_archive.htm
Monday, June 30, 2008 http://groups.yahoo.com/group/aspartameNM/message/1546

former key Hillary Clinton staff Mark Penn and Patti Solis Doyle
use much neurotoxic aspartame Diet Coke – also many other
politicians: Murray 2008.06.30 http://rmforall.blogspot.com/2008_06_01_archive.htm
Monday, June 30, 2008 http://groups.yahoo.com/group/aspartameNM/message/1545 ]

formaldehyde and formic acid in FEMA trailers and other sources
(aspartame, dark wines and liquors, tobacco smoke):
Murray 2008.01.30 http://rmforall.blogspot.com/2008_01_01_archive.htm
Wednesday, January 30, 2008 http://groups.yahoo.com/group/aspartameNM/message/1508

The FEMA trailers give about the same amount of formaldehyde
and formic acid daily as from a quart of dark wine or liquor,
or two quarts (6 12-oz cans) of aspartame diet soda,
from their over 1 tenth gram methanol impurity
(one part in 10,000), which the body quickly makes into
formaldehyde and then formic acid – enough to be the major cause
of "morning after" alcohol hangovers.

Methanol and formaldehyde and formic acid also result from
many fruits and vegetables, tobacco and wood smoke, heater
and vehicle exhaust, household chemicals and cleaners, cosmetics,
and new cars, drapes, carpets, furniture, particleboard,
mobile homes, buildings, leather... so all these sources add up
and interact with many other toxic chemicals.

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: 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
___________________________________________________

"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 127 members, 1,548 posts in a public archive

http://groups.yahoo.com/group/aspartame/messages
group with 1,124 members, 22,792 posts in public archive
___________________________________________________

http://www.newyorker.com/reporting/2008/05/26/080526fa_fact_acocella?currentPage=all

Annals Of Drinking
A Few Too Many
Is there any hope for the hung over?
by Joan Acocella May 26, 2008 themail@newyorker.com

"Wayne Jones, of the Swedish National Laboratory of Forensic Medicine"
[ 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@catholic.ac.kr
Songsin Campus: 02-740-9714 Songsim Campus: 02-2164-4116
Songeui Campus: 02-2164-4114 http://www.cuk.ac.kr/eng/sub055.htm eight hospitals

[ Han-Kyu Lee ]

A hangover is characterized by the unpleasant physical and mental
symptoms that occur between 8 and 16 hours after drinking alcohol.

After inducing experimental hangover in normal individuals,
we measured the methanol concentration prior to
and after alcohol consumption
and we assessed the association between the hangover condition
and the blood methanol level.

A total of 18 normal adult males participated in this study.

They did not have any previous histories of psychiatric
or medical disorders.

The blood ethanol concentration prior to the alcohol intake
(2.26+/-2.08) was not significantly different from that
13 hours after the alcohol consumption (3.12+/-2.38).

However, the difference of methanol concentration
between the day of experiment (prior to the alcohol intake)
and the next day (13 hours after the alcohol intake)
was significant (2.62+/-1.33/l vs. 3.88+/-2.10/l, respectively).

A significant positive correlation was observed
between the changes of blood methanol concentration
and hangover subjective scale score increment when covarying
for the changes of blood ethanol level (r=0.498, p<0.05).

This result suggests the possible correlation of methanol
as well as its toxic metabolite to hangover. PMID: 16318957

[ The toxic metabolite of methanol is formaldehyde, which in turn
partially becomes formic acid – both potent cumulative toxins
that are the actual cause of the toxicity of methanol.]

This study by Jones AW (1987) found next-morning hangover
from red wine with 100 to 150 mg methanol
(9.5 % w/v ethanol, 100 mg/l methanol, 0.01 %).
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).

Pharmacol Toxicol. 1987 Mar; 60(3): 217-20.
Elimination half-life of methanol during hangover.
Jones AW. wayne.jones@RMV.se;
Department of Forensic Toxicology,
University Hospital, SE-581 85 Linkoping, Sweden.

This paper reports the elimination half-life of methanol in human
volunteers.
Experiments were made during the morning after the subjects had
consumed 1000-1500 ml red wine
(9.5 % w/v ethanol, 100 mg/l methanol)
the previous evening. [ 100 to 150 mg methanol ]
The washout of methanol from the body
coincided with the onset of hangover.
The concentrations of ethanol and methanol in blood were
determined indirectly by analysis of end-expired alveolar air.
In the morning when blood-ethanol dropped
below the Km of liver alcohol dehydrogenase (ADH)
of about 100 mg/l (2.2 mM),
the disappearance half-life of ethanol was 21, 22, 18 and 15 min.
in 4 test subjects respectively.
The corresponding elimination half-lives of methanol
were 213, 110, 133 and 142 min. in these same individuals.
The experimental design outlined in this paper can be used
to obtain useful data on elimination kinetics of methanol
in human volunteers without undue ethical limitations.
Circumstantial evidence is presented to link methanol
or its toxic metabolic products, formaldehyde and formic acid,
with the pathogenesis of hangover. PMID: 3588516 ]

"Maria Lucia Souza-Formigoni, a psychobiology researcher at the Federal
University of São Paolo"
[ http://www.newscientist.com/article/dn8901-energy-drink-mixers-give-a-false-sense-of-sobriety.html

Energy drink mixers give a false sense of sobriety
16:52 27 March 2006 NewScientist.com news service, Roxanne Khamsi

'Roseli Boerngen de Lacerda, who studies substance misuse at the Federal
University of Paraná in Curitiba, Brazil'
boerngen@bol.com.br;

Alcohol Clin Exp Res. 2006 Apr; 30(4): 598-605.
Effects of energy drink ingestion on alcohol intoxication.
Ferreira SE, de Mello MT, Pompéia S, de Souza-Formigoni ML.
Department of Psychobiology, Federal University of Sao Paulo (UNIFESP), the
FAPESP fellowship, São Paulo-SP, Brasil.
PMID: 16573577
Sionaldo Eduardo Ferreira
Marco Túlio de Mello
Sabine Pompéia
Maria Lucia Oliveira de Souza-Formigoni, PhD Department of Psychobiology,
Federal University of Sao Paulo (UNIFESP), Rua Botucatu n° 862 1° Andar,
Vila Clementino, São Paulo-SP, Brasil; Fax: +55-11-5572-5092; E-mail:
mlformig@psicobio.epm.br; ]

"Manuela Neuman, a Canadian researcher on alcohol-induced liver damage"
[ manuela@sten.sunnybrook.utoronto.ca;
Manuela Neuman, PhD
Division of Clinical Pharmacology, 2075 Bayview Ave E 242
Sunnybrook HSC, M4N 3M5 Toronto, Ont., Canada
Tel. +1 416 480 6100 ext. 3503, Fax +1 416 480 6025 ]

"Jeffrey Wiese, of Tulane University"
[ MD, Phone Number 504.988.1143 jwiese@tulane.edu ]

"Emil Chiaberi, a co-founder of RU-21's manufacturer, Spirit Sciences, in
California"
[ http://www.spirit-sciences.com/
Mandy Barton, Director of Non-Profit Campaigns, at mb@spirit-sciences.com;
Spirit Sciences USA, Inc
9454 Wilshire Blvd, Suite 600
Beverly Hills, CA 90212
Telephone 310.568.1030 866.556.5577 Facsimile 310.861.5612
General info@spirit-sciences.com;

http://www.ru21.com/
"Dr. Kenneth D Krull, Ph. D., Clinical Biochemist" (I found no leads via
Google )

http://en.wikipedia.org/wiki/RU-21
"Antipokhmelin is a Russian tablet that helps to prevent or overcome the
negative effects of alcohol consumption and hangover. The main ingredient is
succinic acid, also found in amber. It is marketed as RU-21 in the US and
UK. Claims of effectiveness are based primarily on anecdotal evidence, and
there have been no known placebo controlled double blind studies published
in peer reviewed scientific journals.

RU-21 was developed by Prof. Eugene Mayevski at the Institute of Theoretical
and Experimental Biophysics (division of the Russian Academy of Sciences),
where the product was also clinically tested. Further tests were conducted
at the Russian Ministry of Public Health. " ]

"Robert Lindsey, the president of the National Council on Alcoholism and
Drug Dependence"
Robert J. Lindsey
Mr. Lindsey has been in the forefront of the alcoholism and addiction
recovery services community for over 30 years as an employee assistance
professional, Director of Community Relations for the Betty Ford Center, and
Executive Director of a state and a local NCADD Affiliate.
Mr. Lindsey holds a B.A. in Psychology, a Masters of Science in Education
from St. Bonaventure University in New York, and is a Certified Employee
Assistance Professional (CEAP)..... http://www.canys.net/councils.htm
Robert Lindsey, President & CEO NCADD
20 Exchange Place, Suite 2902, New York , NY 10005-3201
Phone: (212) 269-7797 Fax: (212) 269-7510
www.ncadd.org Email: president@ncadd.org; ]

"Robert Swift, an alcohol researcher who teaches at Brown University"
[ http://groups.yahoo.com/group/aspartameNM/message/1047
Avoiding Hangover Hell 2003.12.31 Mark Sherman, AP writer:
Robert Swift, MD [ formaldehyde from methanol in aspartame ]:
Murray 2004.01.16

Robert_Swift_MD@Brown.EDU; joe.schwarcz@mcgill.ca; ]

"Genevieve Ames and her research team at the Prevention Research Center, in
Berkeley"
[ http://sph.berkeley.edu/faculty/ames.html
Genevieve Ames, Ph.D., Adjunct Professor of Medical Anthropology
PHONE: (510) 883-5726 FAX: (510) 644-0594
LOCATION:1995 University Ave., #450, Berkeley, CA 94704
E-MAIL: ames@prev.org;
Research Interests
Anthropology of Health and Healing
Environmental approaches to prevention of substance abuse
Integrating Quantitative and Qualitative Methods
Workplace and alcohol problem prevention ]
____________________________________________________

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: 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.....

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.

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____________________________________________________

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.

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[ Biull Eksp Biol Med. 1986 Nov; 102(11): 573-5.
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[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

British Columbia guidelines against "any drinks with artificial sweeteners"
in January 2008 in school vending machines, stores, cafeterias or
fundraisers – also recently in Ontario and Quebec, Janet Steffenhagen
2007.12.28 Vancouver Sun: Murray 2008.04.10 http://rmforall.blogspot.com/2008_04_01_archive.htm
Thursday, April 10, 2008 http://groups.yahoo.com/group/aspartameNM/message/1537

stevia herbal sweetener to be sold as Truvia (rebiana) by Cargill and
Coca-Cola, if blitz of 12 studies wins FDA approval in 30-90 days: Murray
2008.05.24 http://rmforall.blogspot.com/2008_05_01_archive.htm
Saturday, May 24, 2008 http://groups.yahoo.com/group/aspartameNM/message/1540
____________________________________________________

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."
____________________________________________________

Avoiding formaldehyde allergic reactions in children, aspartame, vitamins,
shampoo, conditioners, hair gel, baby wipes, Sharon E Jacob, MD, Tace
Steele, U. Miami, Pediatric Annals 2007 Jan.: eyelid contact dermatitis, AM
Hill, DV Belsito, 2003 Nov.: Murray 2008.03.27 http://rmforall.blogspot.com/2008_03_01_archive.htm
Thursday, March 27, 2008 http://groups.yahoo.com/group/aspartameNM/message/1532

"It is generally recommended that exposure to products containing
formaldehyde, FRP's, and aspartame (NutraSweet) be avoided
in children."

"Through metabolism, aspartame is converted metabolically
in the liver to methanol,
which is in turn metabolized to formaldehyde. 8"

www.pediatricannalsonline.com/showPdf.asp?rID=21306

Avoiding formaldehyde allergic reactions in children
Pediatric Annals. 2007 Jan.; 36(1): 55-6. PMID: 17269284
Sharon E. Jacob, MD, Director, Contact Dermatitis Clinic,
Dept. of Dermatology and Cutaneous Surgery, U. of Miami,
1295 NW 14th St., Miami, FL 33125, fax 305-243-6191

formaldehyde from many sources, including aspartame, is major cause of
Allergic Contact Dermatitis, SE Jacob, T Steele, G Rodriguez, Skin and Aging
2005 Dec.: Murray 2008.03.27 http://rmforall.blogspot.com/2008_03_01_archive.htm
Thursday, March 27, 2008 http://groups.yahoo.com/group/aspartameNM/message/1533

Sharon E. Jacob, MD
Assistant Professor of Medicine (Dermatology)
University of California, San Diego 200 W. Arbor Drive #8420
San Diego, CA 92103-8420
Tel: 858-552-8585 ×3504 Fax: 305-675-8317
sjacob@contactderm.net;

"For example, diet soda and yogurt containing aspartame
(Nutrasweet), release formaldehyde in their natural biological
degradation.

One of aspartame's metabolites, aspartic acid methyl ester,
is converted to methanol in the body, which is oxidized to
formaldehyde in all organs, including the liver and eyes. 22

Patients with a contact dermatitis to formaldehyde have been seen
to improve once aspartame is avoided. 22

Notably, the case that Hill and Belsito reported had a 6-month
history of eyelid dermatitis that subsided after 1 week of avoiding
diet soda. 22"

"We present a case of a medical student who presented with
erythematous eczematoid plaques on her trunk and legs and
fine vesiculation of her scalp, 3 weeks after starting anatomy class.

Of note, she routinely washed her face and arms after leaving the
anatomy lab, but remained in her scrubs for the rest of the day.

Formaldehyde and Quaternium-15 positive reactions
in the same patient."

"Our patient underscores the importance of appropriate patch
testing and education.
Once we identified the allergy to formaldehyde and quaternium-15,
we provided patient education materials regarding the common and
not-so-common locations of these chemicals and cross-reactors.
We also gave the patient information on avoidance
and safe alternatives (see Table 5).

Fortunately, with technical advances, this student completed the
anatomy section via electronic learning tools.

By avoiding formaldehyde, including anatomy lab, FRP
in her shampoo and cosmetics,
and aspartame in her diet, this patient dramatically improved.

As with all contact dermatitides, the mainstay of treatment for
allergic contact dermatitis is avoidance."

http://www.skinandaging.com/article/5158
Allergen Focus:
Focus on T.R.U.E. Test Allergens #21, 13 and 18:
Formaldehyde and Formaldehyde-Releasing Preservatives
Skin & Aging, ISSN 1096-0120; 13(12) 2005 Dec.: 22-27.
Sharon E. Jacob, M.D.,
Tace Steele, B.A.,
and Georgette Rodriguez, M.D., M.P.H. ]
____________________________________________________

two aspartame (methanol, formaldehyde, formic acid) toxicity research
studies by Resia Pretorius, U. Pretoria, South Africa, debate with JD
Fernstrom: Murray 2008.04.04 2008.05.29 http://rmforall.blogspot.com/2008_04_01_archive.htm
Friday, April 4, 2008 http://groups.yahoo.com/group/aspartameNM/message/1536

http://foodqualitynews.com/news/ng.asp?n=84424-aspartame-sweetener
recent news re E Pretorius aspartame and brain review

Direct and indirect cellular effects of aspartame on the brain.
Humphries P, Pretorius E, Naude H, U. Pretoria, South Africa,
Eur J Clin Nutr. 2007 Aug 8: Murray 2007.08.12 http://groups.yahoo.com/group/aspartameNM/message/1463

"The aim of this study was to discuss the direct and indirect
cellular effects of aspartame on the brain,
and we propose that excessive aspartame ingestion
might be involved in the pathogenesis
of certain mental disorders (DSM-IV-TR 2000)
and also in compromised learning and emotional functioning."

Eur J Clin Nutr. 2007 Aug 8; [Epub ahead of print]
Direct and indirect cellular effects of aspartame on the brain.
Humphries P,
Pretorius E, resia.pretorius@up.ac.za;
Naude H.
[1] Department of Anatomy, University of Pretoria,
Pretoria, Gauteng, South Africa
[2] Department of Anatomy, University of the Limpopo,
South Africa.

The use of the artificial sweetener, aspartame, has long been
contemplated and studied by various researchers, and people are
concerned about its negative effects.

Aspartame is composed of phenylalanine (50%),
aspartic acid (40%) and methanol (10%).

Phenylalanine plays an important role in neurotransmitter regulation,
whereas aspartic acid is also thought to play a role as an excitatory
neurotransmitter in the central nervous system.

Glutamate, asparagines and glutamine are formed from their
precursor, aspartic acid.

Methanol, which forms 10% of the broken down product,
is converted in the body to formate,
which can either be excreted or can give rise to formaldehyde,
diketopiperazine (a carcinogen) and a number of other highly toxic
derivatives.

Previously, it has been reported that consumption of aspartame
could cause neurological and behavioural disturbances in sensitive
individuals.

Headaches, insomnia and seizures are also some of the neurological
effects that have been encountered, and these may be accredited to
changes in regional brain concentrations of catecholamines,
which include norepinephrine, epinephrine and dopamine.

The aim of this study was to discuss the direct and indirect
cellular effects of aspartame on the brain,
and we propose that excessive aspartame ingestion
might be involved in the pathogenesis
of certain mental disorders (DSM-IV-TR 2000)
and also in compromised learning and emotional functioning.

European Journal of Clinical Nutrition advance online publication,
8 August 2007; doi:10.1038/sj.ejcn.1602866.
PMID: 17684524

Keywords: astrocytes; aspartame; neurotransmitters; glutamate;
GABA; serotonin; dopamine; acetylcholine

Received 25 October 2006; revised 26 April 2007;
accepted 27 April 2007
Correspondence: Professor E Pretorius, Department of Anatomy,
University of Pretoria, BMW Building, Dr Savage Street,
PO Box 2034, Pretoria 0001,
Gauteng, South Africa. E-mail: resia.pretorius@up.ac.za

c 2007 Nature Publishing Group,
All rights reserved 0954-3007/07
$30.00 www.nature.com/ejcn

http://groups.yahoo.com/group/aspartameNMmessage/1452
phenylalanine and aspartic acid from low dose aspartame in rabbits
interfere with blood coagulation, Pretorius E and Humphries P,
U. of Pretoria, Ultrastruct Pathol 2007 March: Murray 2007.07.14

" The authors conclude by suggesting that aspartame usage
may interfere with the coagulation process
and might cause delayed fibrin breakup after clot formation.

They suggest this,
as the fibrin networks from aspartame-exposed rabbits
are more complex and dense,
due to the netlike appearance of the minor, thin fibers.

Aspartame usage should possibly be limited
by people on anti-clotting medicine
or those with prone to clot formation. "

Ultrastruct Pathol. 2007 Mar-Apr; 31(2): 77-83.
Ultrastructural changes to rabbit fibrin and platelets
due to aspartame.
Pretorius E,
Humphries P.
Department of Anatomy, Faculty of Medicine,
University of Pretoria, South Africa.
[ Humphries P also at
Department of Anatomy, University of Limpopo.
Medunsa Campus, Garankuwa. South Africa ]

email: E. Pretorius resia.pretorius@up.ac.za
*Correspondence to E. Pretorius,
BMW Building, PO Box 2034,
Faculty of Health Sciences,
University of Pretoria, Pretoria 0001, South Africa

The coagulation process, including thrombin, fibrin,
as well as platelets,
plays an important role in hemostasis,
contributing to the general well-being of humans.

Fibrin formation and platelet activation are delicate processes
that are under the control of many small physiological events.

Any one of these many processes
may be influenced or changed by external factors,
including pharmaceutical or nutritional products, e.g.,
the sweetener aspartame (L-aspartyl-L-phenylalanine methyl ester).

It is known that phenylalanine is present at position P(9)
and aspartate at position P(10)
of the alpha-chain of human fibrinogen,
and plays an important role in the conversion of fibrinogen to fibrin
by the catalyst alpha-thrombin.

The authors investigate the effect of aspartame
on platelet and fibrin ultrastructure,
by using the rabbit animal model
and the scanning electron microscope.

Animals were exposed to 34 mg/kg of aspartame
26x during a 2-month period.

Aspartame-exposed fibrin networks appeared denser,
with a thick matted fine fiber network
covering thick major fibers.

Also, the platelet aggregates appeared more granular
than the globular control platelet aggregates.

The authors conclude by suggesting that aspartame usage
may interfere with the coagulation process
and might cause delayed fibrin breakup after clot formation.

They suggest this,
as the fibrin networks from aspartame-exposed rabbits
are more complex and dense,
due to the netlike appearance of the minor, thin fibers.

Aspartame usage should possibly be limited
by people on anti-clotting medicine
or those with prone to clot formation.
PMID: 17613990
____________________________________________________

http://groups.yahoo.com/group/aspartameNM/message/1459
third study by expert Greek team of neurotoxicity in infant rats by
aspartame (or its parts, methanol, phenylalanine, aspartic acid), KH
Schulpis et al, Food Chem Toxicol 2007.06.16: Murray 2007.08.05

http://groups.yahoo.com/group/aspartameNMmessage/1447
second study by expert Greek team of neurotoxicity in infant rats by
aspartame (or its parts, methanol, phenylalanine, aspartic acid), KH
Schulpis et al, Toxicology 2007.05.18: Murray 2007.07.04

http://groups.yahoo.com/group/aspartameNMmessage/1444
expert Greek group finds aspartame (or its parts, methanol,
phenylalanine, aspartic acid) harm infant rat brain enzyme activity,
KH Schulpis et al, Pharmacol. Res. 2007.05.13:
Murray 2007.06.23

http://groups.yahoo.com/group/aspartameNM/message/939
aspartame (aspartic acid, phenylalanine) binding to DNA:
Karikas July 1998: Murray 2003.01.05 rmforall
Karikas GA, Schulpis KH, Reclos GJ, Kokotos G
Measurement of molecular interaction of aspartame and
its metabolites with DNA. Clin Biochem 1998 Jul; 31(5): 405-7.
Dept. of Chemistry, University of Athens, Greece http://www.chem.uoa.gr gkokotos@atlas.uoa.gr
K.H. Schulpis inchildh@otenet.gr ; G.J. Reclos reklos@otenet.gr

5 recent aspartame reports by S Tsakiris, KH Schulpis, I Simintzi,
with responses to critiques by AG Renwick and
by EB Abegaz, RG Bursey, 2005-2008 2008.03.05

Pharmacological Research 57 (2008) 89-90
Letter to the Editor
Answer to Letter sent to the Editor by
Drs. E. Abegaz and R. Bursey
(Ajinomoto Corporate Services LLC, Washington, USA)
related to Simintzi et al. report published in
Pharmacol Res 2007; 56: 155-9
Letter to the Editor / Pharmacological Research 57 (2008) 89-90

Stylianos Tsakiris a,? stsakir@cc.uoa.gr;
Kleopatra H. Schulpis b inchildh@otenet.gr;
a Department of Experimental Physiology, Medical School,
Athens University, P.O. Box 65257, GR-15401 Athens, Greece

b Inborn Errors of Metabolism Department, Institute of Child
Health, Research Center, Greece
? Corresponding author.
E-mail addresses:
S. Tsakiris stsakir@cc.uoa.gr;
K.H. Schulpis inchildh@otenet.gr;

Pharmacological Research 57 (2008) 87-88
Response to "The effect of aspartame on the acetylcholinesterase
activity in hippocampal homogenates of suckling rats"
by Simintzi et al.

Eyassu G. Abegaz ?
Robert G. Bursey
Ajinomoto Corporate Services LLC,
Scientific & Regulatory Affairs,
1120 Connecticut Ave., N.W., Suite 1010,
Washington, DC 20036, United States

? Corresponding author. Tel.: +1 202 457 0284;
fax: +1 202 457 0107.
E-mail addresses: abegazee@ajiusa.com; (E.G. Abegaz),
burseyb@ajiusa.com; (R.G. Bursey)

Keywords:
Aspartame; Aspartate; Phenylalanine; Methanol; AChE activity

Tsakiris S, Schulpis KH.
Answer to letter sent by Professor A.G. Renwick
(University of Southampton, UK)
related to Simintzi et al. report published in Food and Chemical
Toxicology 2007; 45(12): 2397-401.
Food Chem Toxicol. 2008 Mar; 46(3): 1208-9.
Epub 2007 Oct 25. No abstract available. PMID: 18054419
doi:10.1016/j.fct.2007.10.016
Copyright © 2007 Elsevier Ltd All rights reserved.

Renwick AG.
The effect of aspartame metabolites on the suckling rat frontal cortex
acetylcholinesterase. An in vitro study. By I. Simintzi, K.H. Schulpis,
P. Angelogianni, C. Liapi and S. Tsakiris.
Food Chem Toxicol. 2008 Mar; 46(3): 1206-7.
Epub 2007 Oct 26. No abstract available. PMID: 18061330

1: Simintzi I, Schulpis KH, Angelogianni P, Liapi C, Tsakiris S.
The effect of aspartame metabolites on the suckling rat frontal cortex
acetylcholinesterase. An in vitro study.
Food Chem Toxicol. 2007 Dec;45(12):2397-401.
Epub 2007 Jun 16. PMID: 17673349

2: Simintzi I, Schulpis KH, Angelogianni P, Liapi C, Tsakiris S.
L-Cysteine and glutathione restore the reduction of rat
hippocampal Na+, K+-ATPase activity
induced by aspartame metabolites.
Toxicology. 2007 Jul 31;237(1-3):177-83.
Epub 2007 May 18. PMID: 17602817

3: Simintzi I, Schulpis KH, Angelogianni P, Liapi C, Tsakiris S.
The effect of aspartame on acetylcholinesterase activity in
hippocampal homogenates of suckling rats.
Pharmacol Res. 2007 Aug;56(2):155-9.
Epub 2007 May 13. PMID: 17580119

4: Schulpis KH, Papassotiriou I, Parthimos T, Tsakiris T, Tsakiris S.
The effect of L-cysteine and glutathione
on inhibition of Na+, K+-ATPase activity by aspartame metabolites
in human erythrocyte membrane.
Eur J Clin Nutr. 2006 May;60(5):593-7. PMID: 16391576

5: Tsakiris S, Giannoulia-Karantana A, Simintzi I, Schulpis KH.
The effect of aspartame metabolites on human erythrocyte
membrane acetylcholinesterase activity.
Pharmacol Res. 2006 Jan;53(1):1-5.
Epub 2005 Aug 29. PMID: 16129618
____________________________________________________

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
____________________________________________________

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
____________________________________________________

two detailed critiques of industry affiliations and biased science in 99
page review with 415 references by BA Magnuson, GA Burdock
and 8 more, Critical Reviews in Toxicology, 2007 Sept.: Mark D
Gold 13 page: also Rich Murray 2007.09.15: 2008.03.24 http://rmforall.blogspot.com/2008_03_01_archive.htm
Monday, March 24, 2008 http://groups.yahoo.com/group/aspartameNM/message/1531

"Nearly every section of the Magnuson (2007) review has research
that is misrepresented
and/or crucial pieces of information are left out.

In addition to the misrepresentation of the research,
readers (including medical professionals) are often not told that
this review was funded by the aspartame manufacturer, Ajinomoto,
and the reviewers had enormous conflicts of interest."
____________________________________________________

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
____________________________________________________

re "A Few too Many", Joan Acocella, The New Yorker, long review of hangover
research 2008.05.26 – same levels of formaldehyde and formic acid in FEMA
trailers and other sources (aspartame, dark wines and liquors, tobacco
smoke): Murray 2008.06.05 http://rmforall.blogspot.com/2008_06_01_archive.htm
Thursday, June 5, 2008 http://groups.yahoo.com/group/aspartameNM/message/1541

[ See also:
There really is no controversy, Adrienne Samuels PhD, letter re
evident toxicity of aspartame EJCN 2008.06.11:
Murray 2008.06.30 http://rmforall.blogspot.com/2008_06_01_archive.htm
Monday, June 30, 2008 http://groups.yahoo.com/group/aspartameNM/message/1546

former key Hillary Clinton staff Mark Penn and Patti Solis Doyle
use much neurotoxic aspartame Diet Coke – also many other
politicians: Murray 2008.06.30 http://rmforall.blogspot.com/2008_06_01_archive.htm
Monday, June 30, 2008 http://groups.yahoo.com/group/aspartameNM/message/1545 ]

formaldehyde and formic acid in FEMA trailers and other sources
(aspartame, dark wines and liquors, tobacco smoke):
Murray 2008.01.30 http://rmforall.blogspot.com/2008_01_01_archive.htm
Wednesday, January 30, 2008 http://groups.yahoo.com/group/aspartameNM/message/1508

The FEMA trailers give about the same amount of formaldehyde
and formic acid daily as from a quart of dark wine or liquor,
or two quarts (6 12-oz cans) of aspartame diet soda,
from their over 1 tenth gram methanol impurity
(one part in 10,000), which the body quickly makes into
formaldehyde and then formic acid – enough to be the major cause
of "morning after" alcohol hangovers.

Methanol and formaldehyde and formic acid also result from
many fruits and vegetables, tobacco and wood smoke, heater
and vehicle exhaust, household chemicals and cleaners, cosmetics,
and new cars, drapes, carpets, furniture, particleboard,
mobile homes, buildings, leather... so all these sources add up
and interact with many other toxic chemicals.

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: 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
___________________________________________________

"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 127 members, 1,548 posts in a public archive

http://groups.yahoo.com/group/aspartame/messages
group with 1,124 members, 22,792 posts in public archive
___________________________________________________

http://www.newyorker.com/reporting/2008/05/26/080526fa_fact_acocella?currentPage=all

Annals Of Drinking
A Few Too Many
Is there any hope for the hung over?
by Joan Acocella May 26, 2008 themail@newyorker.com

"Wayne Jones, of the Swedish National Laboratory of Forensic Medicine"
[ 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@catholic.ac.kr
Songsin Campus: 02-740-9714 Songsim Campus: 02-2164-4116
Songeui Campus: 02-2164-4114 http://www.cuk.ac.kr/eng/sub055.htm eight hospitals

[ Han-Kyu Lee ]

A hangover is characterized by the unpleasant physical and mental
symptoms that occur between 8 and 16 hours after drinking alcohol.

After inducing experimental hangover in normal individuals,
we measured the methanol concentration prior to
and after alcohol consumption
and we assessed the association between the hangover condition
and the blood methanol level.

A total of 18 normal adult males participated in this study.

They did not have any previous histories of psychiatric
or medical disorders.

The blood ethanol concentration prior to the alcohol intake
(2.26+/-2.08) was not significantly different from that
13 hours after the alcohol consumption (3.12+/-2.38).

However, the difference of methanol concentration
between the day of experiment (prior to the alcohol intake)
and the next day (13 hours after the alcohol intake)
was significant (2.62+/-1.33/l vs. 3.88+/-2.10/l, respectively).

A significant positive correlation was observed
between the changes of blood methanol concentration
and hangover subjective scale score increment when covarying
for the changes of blood ethanol level (r=0.498, p<0.05).

This result suggests the possible correlation of methanol
as well as its toxic metabolite to hangover. PMID: 16318957

[ The toxic metabolite of methanol is formaldehyde, which in turn
partially becomes formic acid – both potent cumulative toxins
that are the actual cause of the toxicity of methanol.]

This study by Jones AW (1987) found next-morning hangover
from red wine with 100 to 150 mg methanol
(9.5 % w/v ethanol, 100 mg/l methanol, 0.01 %).
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).

Pharmacol Toxicol. 1987 Mar; 60(3): 217-20.
Elimination half-life of methanol during hangover.
Jones AW. wayne.jones@RMV.se;
Department of Forensic Toxicology,
University Hospital, SE-581 85 Linkoping, Sweden.

This paper reports the elimination half-life of methanol in human
volunteers.
Experiments were made during the morning after the subjects had
consumed 1000-1500 ml red wine
(9.5 % w/v ethanol, 100 mg/l methanol)
the previous evening. [ 100 to 150 mg methanol ]
The washout of methanol from the body
coincided with the onset of hangover.
The concentrations of ethanol and methanol in blood were
determined indirectly by analysis of end-expired alveolar air.
In the morning when blood-ethanol dropped
below the Km of liver alcohol dehydrogenase (ADH)
of about 100 mg/l (2.2 mM),
the disappearance half-life of ethanol was 21, 22, 18 and 15 min.
in 4 test subjects respectively.
The corresponding elimination half-lives of methanol
were 213, 110, 133 and 142 min. in these same individuals.
The experimental design outlined in this paper can be used
to obtain useful data on elimination kinetics of methanol
in human volunteers without undue ethical limitations.
Circumstantial evidence is presented to link methanol
or its toxic metabolic products, formaldehyde and formic acid,
with the pathogenesis of hangover. PMID: 3588516 ]

"Maria Lucia Souza-Formigoni, a psychobiology researcher at the Federal
University of São Paolo"
[ http://www.newscientist.com/article/dn8901-energy-drink-mixers-give-a-false-sense-of-sobriety.html

Energy drink mixers give a false sense of sobriety
16:52 27 March 2006 NewScientist.com news service, Roxanne Khamsi

'Roseli Boerngen de Lacerda, who studies substance misuse at the Federal
University of Paraná in Curitiba, Brazil'
boerngen@bol.com.br;

Alcohol Clin Exp Res. 2006 Apr; 30(4): 598-605.
Effects of energy drink ingestion on alcohol intoxication.
Ferreira SE, de Mello MT, Pompéia S, de Souza-Formigoni ML.
Department of Psychobiology, Federal University of Sao Paulo (UNIFESP), the
FAPESP fellowship, São Paulo-SP, Brasil.
PMID: 16573577
Sionaldo Eduardo Ferreira
Marco Túlio de Mello
Sabine Pompéia
Maria Lucia Oliveira de Souza-Formigoni, PhD Department of Psychobiology,
Federal University of Sao Paulo (UNIFESP), Rua Botucatu n° 862 1° Andar,
Vila Clementino, São Paulo-SP, Brasil; Fax: +55-11-5572-5092; E-mail:
mlformig@psicobio.epm.br; ]

"Manuela Neuman, a Canadian researcher on alcohol-induced liver damage"
[ manuela@sten.sunnybrook.utoronto.ca;
Manuela Neuman, PhD
Division of Clinical Pharmacology, 2075 Bayview Ave E 242
Sunnybrook HSC, M4N 3M5 Toronto, Ont., Canada
Tel. +1 416 480 6100 ext. 3503, Fax +1 416 480 6025 ]

"Jeffrey Wiese, of Tulane University"
[ MD, Phone Number 504.988.1143 jwiese@tulane.edu ]

"Emil Chiaberi, a co-founder of RU-21's manufacturer, Spirit Sciences, in
California"
[ http://www.spirit-sciences.com/
Mandy Barton, Director of Non-Profit Campaigns, at mb@spirit-sciences.com;
Spirit Sciences USA, Inc
9454 Wilshire Blvd, Suite 600
Beverly Hills, CA 90212
Telephone 310.568.1030 866.556.5577 Facsimile 310.861.5612
General info@spirit-sciences.com;

http://www.ru21.com/
"Dr. Kenneth D Krull, Ph. D., Clinical Biochemist" (I found no leads via
Google )

http://en.wikipedia.org/wiki/RU-21
"Antipokhmelin is a Russian tablet that helps to prevent or overcome the
negative effects of alcohol consumption and hangover. The main ingredient is
succinic acid, also found in amber. It is marketed as RU-21 in the US and
UK. Claims of effectiveness are based primarily on anecdotal evidence, and
there have been no known placebo controlled double blind studies published
in peer reviewed scientific journals.

RU-21 was developed by Prof. Eugene Mayevski at the Institute of Theoretical
and Experimental Biophysics (division of the Russian Academy of Sciences),
where the product was also clinically tested. Further tests were conducted
at the Russian Ministry of Public Health. " ]

"Robert Lindsey, the president of the National Council on Alcoholism and
Drug Dependence"
Robert J. Lindsey
Mr. Lindsey has been in the forefront of the alcoholism and addiction
recovery services community for over 30 years as an employee assistance
professional, Director of Community Relations for the Betty Ford Center, and
Executive Director of a state and a local NCADD Affiliate.
Mr. Lindsey holds a B.A. in Psychology, a Masters of Science in Education
from St. Bonaventure University in New York, and is a Certified Employee
Assistance Professional (CEAP)..... http://www.canys.net/councils.htm
Robert Lindsey, President & CEO NCADD
20 Exchange Place, Suite 2902, New York , NY 10005-3201
Phone: (212) 269-7797 Fax: (212) 269-7510
www.ncadd.org Email: president@ncadd.org; ]

"Robert Swift, an alcohol researcher who teaches at Brown University"
[ http://groups.yahoo.com/group/aspartameNM/message/1047
Avoiding Hangover Hell 2003.12.31 Mark Sherman, AP writer:
Robert Swift, MD [ formaldehyde from methanol in aspartame ]:
Murray 2004.01.16

Robert_Swift_MD@Brown.EDU; joe.schwarcz@mcgill.ca; ]

"Genevieve Ames and her research team at the Prevention Research Center, in
Berkeley"
[ http://sph.berkeley.edu/faculty/ames.html
Genevieve Ames, Ph.D., Adjunct Professor of Medical Anthropology
PHONE: (510) 883-5726 FAX: (510) 644-0594
LOCATION:1995 University Ave., #450, Berkeley, CA 94704
E-MAIL: ames@prev.org;
Research Interests
Anthropology of Health and Healing
Environmental approaches to prevention of substance abuse
Integrating Quantitative and Qualitative Methods
Workplace and alcohol problem prevention ]
____________________________________________________

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: 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.....

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.

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____________________________________________________

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

#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

British Columbia guidelines against "any drinks with artificial sweeteners"
in January 2008 in school vending machines, stores, cafeterias or
fundraisers – also recently in Ontario and Quebec, Janet Steffenhagen
2007.12.28 Vancouver Sun: Murray 2008.04.10 http://rmforall.blogspot.com/2008_04_01_archive.htm
Thursday, April 10, 2008 http://groups.yahoo.com/group/aspartameNM/message/1537

stevia herbal sweetener to be sold as Truvia (rebiana) by Cargill and
Coca-Cola, if blitz of 12 studies wins FDA approval in 30-90 days: Murray
2008.05.24 http://rmforall.blogspot.com/2008_05_01_archive.htm
Saturday, May 24, 2008 http://groups.yahoo.com/group/aspartameNM/message/1540
____________________________________________________

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."
____________________________________________________

Avoiding formaldehyde allergic reactions in children, aspartame, vitamins,
shampoo, conditioners, hair gel, baby wipes, Sharon E Jacob, MD, Tace
Steele, U. Miami, Pediatric Annals 2007 Jan.: eyelid contact dermatitis, AM
Hill, DV Belsito, 2003 Nov.: Murray 2008.03.27 http://rmforall.blogspot.com/2008_03_01_archive.htm
Thursday, March 27, 2008 http://groups.yahoo.com/group/aspartameNM/message/1532

"It is generally recommended that exposure to products containing
formaldehyde, FRP's, and aspartame (NutraSweet) be avoided
in children."

"Through metabolism, aspartame is converted metabolically
in the liver to methanol,
which is in turn metabolized to formaldehyde. 8"

www.pediatricannalsonline.com/showPdf.asp?rID=21306

Avoiding formaldehyde allergic reactions in children
Pediatric Annals. 2007 Jan.; 36(1): 55-6. PMID: 17269284
Sharon E. Jacob, MD, Director, Contact Dermatitis Clinic,
Dept. of Dermatology and Cutaneous Surgery, U. of Miami,
1295 NW 14th St., Miami, FL 33125, fax 305-243-6191

formaldehyde from many sources, including aspartame, is major cause of
Allergic Contact Dermatitis, SE Jacob, T Steele, G Rodriguez, Skin and Aging
2005 Dec.: Murray 2008.03.27 http://rmforall.blogspot.com/2008_03_01_archive.htm
Thursday, March 27, 2008 http://groups.yahoo.com/group/aspartameNM/message/1533

Sharon E. Jacob, MD
Assistant Professor of Medicine (Dermatology)
University of California, San Diego 200 W. Arbor Drive #8420
San Diego, CA 92103-8420
Tel: 858-552-8585 ×3504 Fax: 305-675-8317
sjacob@contactderm.net;

"For example, diet soda and yogurt containing aspartame
(Nutrasweet), release formaldehyde in their natural biological
degradation.

One of aspartame's metabolites, aspartic acid methyl ester,
is converted to methanol in the body, which is oxidized to
formaldehyde in all organs, including the liver and eyes. 22

Patients with a contact dermatitis to formaldehyde have been seen
to improve once aspartame is avoided. 22

Notably, the case that Hill and Belsito reported had a 6-month
history of eyelid dermatitis that subsided after 1 week of avoiding
diet soda. 22"

"We present a case of a medical student who presented with
erythematous eczematoid plaques on her trunk and legs and
fine vesiculation of her scalp, 3 weeks after starting anatomy class.

Of note, she routinely washed her face and arms after leaving the
anatomy lab, but remained in her scrubs for the rest of the day.

Formaldehyde and Quaternium-15 positive reactions
in the same patient."

"Our patient underscores the importance of appropriate patch
testing and education.
Once we identified the allergy to formaldehyde and quaternium-15,
we provided patient education materials regarding the common and
not-so-common locations of these chemicals and cross-reactors.
We also gave the patient information on avoidance

and safe alternatives (see Table 5).

Fortunately, with technical advances, this student completed the
anatomy section via electronic learning tools.

By avoiding formaldehyde, including anatomy lab, FRP
in her shampoo and cosmetics,
and aspartame in her diet, this patient dramatically improved.

As with all contact dermatitides, the mainstay of treatment for
allergic contact dermatitis is avoidance."

http://www.skinandaging.com/article/5158
Allergen Focus:
Focus on T.R.U.E. Test Allergens #21, 13 and 18:
Formaldehyde and Formaldehyde-Releasing Preservatives
Skin & Aging, ISSN 1096-0120; 13(12) 2005 Dec.: 22-27.
Sharon E. Jacob, M.D.,
Tace Steele, B.A.,
and Georgette Rodriguez, M.D., M.P.H. ]
____________________________________________________

two aspartame (methanol, formaldehyde, formic acid) toxicity research
studies by Resia Pretorius, U. Pretoria, South Africa, debate with JD
Fernstrom: Murray 2008.04.04 2008.05.29 http://rmforall.blogspot.com/2008_04_01_archive.htm
Friday, April 4, 2008 http://groups.yahoo.com/group/aspartameNM/message/1536

http://foodqualitynews.com/news/ng.asp?n=84424-aspartame-sweetener
recent news re E Pretorius aspartame and brain review

Direct and indirect cellular effects of aspartame on the brain.
Humphries P, Pretorius E, Naude H, U. Pretoria, South Africa,
Eur J Clin Nutr. 2007 Aug 8: Murray 2007.08.12 http://groups.yahoo.com/group/aspartameNM/message/1463

"The aim of this study was to discuss the direct and indirect
cellular effects of aspartame on the brain,
and we propose that excessive aspartame ingestion
might be involved in the pathogenesis
of certain mental disorders (DSM-IV-TR 2000)
and also in compromised learning and emotional functioning."

Eur J Clin Nutr. 2007 Aug 8; ( Epub ahead of print )
Direct and indirect cellular effects of aspartame on the brain.
Humphries P,
Pretorius E, resia.pretorius@up.ac.za;
Naude H.
[1] Department of Anatomy, University of Pretoria,
Pretoria, Gauteng, South Africa
[2] Department of Anatomy, University of the Limpopo,
South Africa.

The use of the artificial sweetener, aspartame, has long been
contemplated and studied by various researchers, and people are
concerned about its negative effects.

Aspartame is composed of phenylalanine (50%),
aspartic acid (40%) and methanol (10%).

Phenylalanine plays an important role in neurotransmitter regulation,
whereas aspartic acid is also thought to play a role as an excitatory
neurotransmitter in the central nervous system.

Glutamate, asparagines and glutamine are formed from their
precursor, aspartic acid.

Methanol, which forms 10% of the broken down product,
is converted in the body to formate,
which can either be excreted or can give rise to formaldehyde,
diketopiperazine (a carcinogen) and a number of other highly toxic
derivatives.

Previously, it has been reported that consumption of aspartame
could cause neurological and behavioural disturbances in sensitive
individuals.

Headaches, insomnia and seizures are also some of the neurological
effects that have been encountered, and these may be accredited to
changes in regional brain concentrations of catecholamines,
which include norepinephrine, epinephrine and dopamine.

The aim of this study was to discuss the direct and indirect
cellular effects of aspartame on the brain,
and we propose that excessive aspartame ingestion
might be involved in the pathogenesis
of certain mental disorders (DSM-IV-TR 2000)
and also in compromised learning and emotional functioning.

European Journal of Clinical Nutrition advance online publication,
8 August 2007; doi:10.1038/sj.ejcn.1602866.
PMID: 17684524

Keywords: astrocytes; aspartame; neurotransmitters; glutamate;
GABA; serotonin; dopamine; acetylcholine

Received 25 October 2006; revised 26 April 2007;
accepted 27 April 2007
Correspondence: Professor E Pretorius, Department of Anatomy,
University of Pretoria, BMW Building, Dr Savage Street,
PO Box 2034, Pretoria 0001,
Gauteng, South Africa. E-mail: resia.pretorius@up.ac.za

c 2007 Nature Publishing Group,
All rights reserved 0954-3007/07
$30.00 www.nature.com/ejcn

http://groups.yahoo.com/group/aspartameNMmessage/1452
phenylalanine and aspartic acid from low dose aspartame in rabbits
interfere with blood coagulation, Pretorius E and Humphries P,
U. of Pretoria, Ultrastruct Pathol 2007 March: Murray 2007.07.14

" The authors conclude by suggesting that aspartame usage
may interfere with the coagulation process
and might cause delayed fibrin breakup after clot formation.

They suggest this,
as the fibrin networks from aspartame-exposed rabbits
are more complex and dense,
due to the netlike appearance of the minor, thin fibers.

Aspartame usage should possibly be limited
by people on anti-clotting medicine
or those with prone to clot formation. "

Ultrastruct Pathol. 2007 Mar-Apr; 31(2): 77-83.
Ultrastructural changes to rabbit fibrin and platelets
due to aspartame.
Pretorius E,
Humphries P.
Department of Anatomy, Faculty of Medicine,
University of Pretoria, South Africa.
[ Humphries P also at
Department of Anatomy, University of Limpopo.
Medunsa Campus, Garankuwa. South Africa ]

email: E. Pretorius resia.pretorius@up.ac.za
*Correspondence to E. Pretorius,
BMW Building, PO Box 2034,
Faculty of Health Sciences,
University of Pretoria, Pretoria 0001, South Africa

The coagulation process, including thrombin, fibrin,
as well as platelets,
plays an important role in hemostasis,
contributing to the general well-being of humans.

Fibrin formation and platelet activation are delicate processes
that are under the control of many small physiological events.

Any one of these many processes
may be influenced or changed by external factors,
including pharmaceutical or nutritional products, e.g.,
the sweetener aspartame (L-aspartyl-L-phenylalanine methyl ester).

It is known that phenylalanine is present at position P(9)
and aspartate at position P(10)
of the alpha-chain of human fibrinogen,
and plays an important role in the conversion of fibrinogen to fibrin
by the catalyst alpha-thrombin.

The authors investigate the effect of aspartame
on platelet and fibrin ultrastructure,
by using the rabbit animal model
and the scanning electron microscope.

Animals were exposed to 34 mg/kg of aspartame
26x during a 2-month period.

Aspartame-exposed fibrin networks appeared denser,
with a thick matted fine fiber network
covering thick major fibers.

Also, the platelet aggregates appeared more granular
than the globular control platelet aggregates.

The authors conclude by suggesting that aspartame usage
may interfere with the coagulation process
and might cause delayed fibrin breakup after clot formation.

They suggest this,
as the fibrin networks from aspartame-exposed rabbits
are more complex and dense,
due to the netlike appearance of the minor, thin fibers.

Aspartame usage should possibly be limited
by people on anti-clotting medicine
or those with prone to clot formation.
PMID: 17613990
____________________________________________________

http://groups.yahoo.com/group/aspartameNM/message/1459
third study by expert Greek team of neurotoxicity in infant rats by
aspartame (or its parts, methanol, phenylalanine, aspartic acid), KH
Schulpis et al, Food Chem Toxicol 2007.06.16: Murray 2007.08.05

http://groups.yahoo.com/group/aspartameNMmessage/1447
second study by expert Greek team of neurotoxicity in infant rats by
aspartame (or its parts, methanol, phenylalanine, aspartic acid), KH
Schulpis et al, Toxicology 2007.05.18: Murray 2007.07.04

http://groups.yahoo.com/group/aspartameNMmessage/1444
expert Greek group finds aspartame (or its parts, methanol,
phenylalanine, aspartic acid) harm infant rat brain enzyme activity,
KH Schulpis et al, Pharmacol. Res. 2007.05.13:
Murray 2007.06.23

http://groups.yahoo.com/group/aspartameNM/message/939
aspartame (aspartic acid, phenylalanine) binding to DNA:
Karikas July 1998: Murray 2003.01.05 rmforall
Karikas GA, Schulpis KH, Reclos GJ, Kokotos G
Measurement of molecular interaction of aspartame and
its metabolites with DNA. Clin Biochem 1998 Jul; 31(5): 405-7.
Dept. of Chemistry, University of Athens, Greece http://www.chem.uoa.gr gkokotos@atlas.uoa.gr
K.H. Schulpis inchildh@otenet.gr ; G.J. Reclos reklos@otenet.gr

5 recent aspartame reports by S Tsakiris, KH Schulpis, I Simintzi,
with responses to critiques by AG Renwick and
by EB Abegaz, RG Bursey, 2005-2008 2008.03.05

Pharmacological Research 57 (2008) 89-90
Letter to the Editor
Answer to Letter sent to the Editor by
Drs. E. Abegaz and R. Bursey
(Ajinomoto Corporate Services LLC, Washington, USA)
related to Simintzi et al. report published in
Pharmacol Res 2007; 56: 155-9
Letter to the Editor / Pharmacological Research 57 (2008) 89-90

Stylianos Tsakiris a,? stsakir@cc.uoa.gr;
Kleopatra H. Schulpis b inchildh@otenet.gr;
a Department of Experimental Physiology, Medical School,
Athens University, P.O. Box 65257, GR-15401 Athens, Greece

b Inborn Errors of Metabolism Department, Institute of Child
Health, Research Center, Greece
? Corresponding author.
E-mail addresses:
S. Tsakiris stsakir@cc.uoa.gr;
K.H. Schulpis inchildh@otenet.gr;

Pharmacological Research 57 (2008) 87-88
Response to "The effect of aspartame on the acetylcholinesterase
activity in hippocampal homogenates of suckling rats"
by Simintzi et al.

Eyassu G. Abegaz ?
Robert G. Bursey
Ajinomoto Corporate Services LLC,
Scientific & Regulatory Affairs,
1120 Connecticut Ave., N.W., Suite 1010,
Washington, DC 20036, United States

? Corresponding author. Tel.: +1 202 457 0284;
fax: +1 202 457 0107.
E-mail addresses: abegazee@ajiusa.com; (E.G. Abegaz),
burseyb@ajiusa.com; (R.G. Bursey)

Keywords:
Aspartame; Aspartate; Phenylalanine; Methanol; AChE activity

Tsakiris S, Schulpis KH.
Answer to letter sent by Professor A.G. Renwick
(University of Southampton, UK)
related to Simintzi et al. report published in Food and Chemical
Toxicology 2007; 45(12): 2397-401.
Food Chem Toxicol. 2008 Mar; 46(3): 1208-9.
Epub 2007 Oct 25. No abstract available. PMID: 18054419
doi:10.1016/j.fct.2007.10.016
Copyright © 2007 Elsevier Ltd All rights reserved.

Renwick AG.
The effect of aspartame metabolites on the suckling rat frontal cortex
acetylcholinesterase. An in vitro study. By I. Simintzi, K.H. Schulpis,
P. Angelogianni, C. Liapi and S. Tsakiris.
Food Chem Toxicol. 2008 Mar; 46(3): 1206-7.
Epub 2007 Oct 26. No abstract available. PMID: 18061330

1: Simintzi I, Schulpis KH, Angelogianni P, Liapi C, Tsakiris S.
The effect of aspartame metabolites on the suckling rat frontal cortex
acetylcholinesterase. An in vitro study.
Food Chem Toxicol. 2007 Dec;45(12):2397-401.
Epub 2007 Jun 16. PMID: 17673349

2: Simintzi I, Schulpis KH, Angelogianni P, Liapi C, Tsakiris S.
L-Cysteine and glutathione restore the reduction of rat
hippocampal Na+, K+-ATPase activity
induced by aspartame metabolites.
Toxicology. 2007 Jul 31;237(1-3):177-83.
Epub 2007 May 18. PMID: 17602817

3: Simintzi I, Schulpis KH, Angelogianni P, Liapi C, Tsakiris S.
The effect of aspartame on acetylcholinesterase activity in
hippocampal homogenates of suckling rats.
Pharmacol Res. 2007 Aug;56(2):155-9.
Epub 2007 May 13. PMID: 17580119

4: Schulpis KH, Papassotiriou I, Parthimos T, Tsakiris T, Tsakiris S.
The effect of L-cysteine and glutathione
on inhibition of Na+, K+-ATPase activity by aspartame metabolites
in human erythrocyte membrane.
Eur J Clin Nutr. 2006 May;60(5):593-7. PMID: 16391576

5: Tsakiris S, Giannoulia-Karantana A, Simintzi I, Schulpis KH.
The effect of aspartame metabolites on human erythrocyte
membrane acetylcholinesterase activity.
Pharmacol Res. 2006 Jan;53(1):1-5.
Epub 2005 Aug 29. PMID: 16129618
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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
____________________________________________________

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
____________________________________________________

two detailed critiques of industry affiliations and biased science in 99
page review with 415 references by BA Magnuson, GA Burdock
and 8 more, Critical Reviews in Toxicology, 2007 Sept.: Mark D
Gold 13 page: also Rich Murray 2007.09.15: 2008.03.24 http://rmforall.blogspot.com/2008_03_01_archive.htm
Monday, March 24, 2008 http://groups.yahoo.com/group/aspartameNM/message/1531

"Nearly every section of the Magnuson (2007) review has research
that is misrepresented
and/or crucial pieces of information are left out.

In addition to the misrepresentation of the research,
readers (including medical professionals) are often not told that
this review was funded by the aspartame manufacturer, Ajinomoto,
and the reviewers had enormous conflicts of interest."
____________________________________________________

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
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