bias, omissions, incuriosity = opportunity, aspartame safety evaluation,
Magnuson BA, Burdock GA, Williams GM, 7 more, 2007 Sept, Ajinomoto
funded 98 pages html [$ 32 781888262_content.pdf]: Murray 2007.09.15
http://groups.yahoo.com/group/aspartameNM/message/1472

bmagnuso@...,info@...,gburdock@...,jdoull@...\
,gmarsh@...,mwpariza@...,spencer@...,
bwaddell@...,R.Walker@...,preston.julian@...,David.Kirkl\
and@...,gary_williams@...,


Herein I give selections from ASE, along with critical comments and
notes in square brackets, along with their 415 references.


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

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

http://groups.yahoo.com/group/aspartameNM/messages
group with 82 members, 1,472 posts in a public,
searchable archive http://RMForAll.blogspot.com
////////////////////////////////////////////////////////////



Unhee Lim 1,
Amy F. Subar 2, subara@...,
Traci Mouw 1,
Patricia Hartge 1,
Lindsay M. Morton 1,
Rachael Stolzenberg-Solomon 1,
David Campbell 3,
Albert R. Hollenbeck 4
and Arthur Schatzkin 1

1 Division of Cancer Epidemiology and Genetics,

2 Division of Cancer Control and Population Sciences, National Cancer
Institute, NIH, Department of Health and Human Services;

3 Information Management Services, Inc., Rockville, Maryland; and

4 AARP, Washington, District of Columbia

Requests for reprints: Amy Subar,
Division of Cancer Control and Population Sciences,
National Cancer Institute,
6130 Executive Boulevard, EPN 4005, Rockville, MD 20852-7344.
Phone: 301-594-0831; Fax: 301-435-3710. E-mail: subara@...

http://cebp.aacrjournals.org/cgi/content/full/15/9/1654 free full text

BACKGROUND:
In a few animal experiments, aspartame has been linked to hematopoietic
and brain cancers.

Most animal studies have found no increase in the risk of these or other
cancers.

Data on humans are sparse for either cancer.

Concern lingers regarding this widely used artificial sweetener.

OBJECTIVE:
We investigated prospectively whether aspartame consumption is
associated with the risk of hematopoietic cancers or gliomas (malignant
brain cancer).

METHODS:
We examined 285,079 men and 188,905 women ages 50 to 71 years in the
NIH-AARP Diet and Health Study cohort

Daily aspartame intake was derived from responses to a baseline
self-administered food frequency questionnaire that queried consumption
of four aspartame-containing beverages (soda, fruit drinks, sweetened
iced tea, and aspartame added to hot coffee and tea) during the past year.

Histologically confirmed incident cancers were identified from eight
state cancer registries.

Multivariable-adjusted relative risks (RR) and 95% confidence intervals
(CI) were estimated using Cox proportional hazards regression that
adjusted for age, sex, ethnicity, body mass index, and history of diabetes.

RESULTS:
During over 5 years of follow-up (1995-2000), 1,888 hematopoietic
cancers and 315 malignant gliomas were ascertained.

Higher levels of aspartame intake were not associated with the risk of
overall hematopoietic cancer
(RR for >/=600 mg/d, 0.98; 95% CI, 0.76-1.27),
glioma (RR for >/=400 mg/d, 0.73; 95% CI, 0.46-1.15;
P for inverse linear trend = 0.05),
or their subtypes in men and women.

CONCLUSIONS:
Our findings do not support the hypothesis that aspartame increases
hematopoietic or brain cancer risk. PMID: 16985027

"We cannot exclude the possibility that higher aspartame consumption
than that observed in this study may be associated with an elevated risk
of hematopoietic or brain cancers.

In the laboratory study with positive findings, animals were fed doses
starting from 4 mg up to 5,000 mg per kg body weight.

Significantly elevated lymphomas and leukemias were observed in female
rats fed 20 mg of aspartame and higher (e.g., 1,200 mg for humans
weighing 60 kg or 132 lb; refs. 13, 14).

The reported aspartame intake in our data ranged from 0 to 3,400 mg/d
with sparse numbers in the upper intake categories
(<1% consuming ≥1,200 mg/d).

However, we did not detect any increase in risk estimates in the highest
categories (>1,200 or 2,000 mg/d, which is equivalent to ~7 to 11 cans
of soft drinks daily) compared with the lowest categories,
and the associations were similarly null in both men and women."

[ This is the first good data about the percentage of aspartame users
who use over 6 cans daily.

About 1% of 473,984 is 4700 people, with a peak intake of 17 cans daily.

It would be worthwhile to investigate a wide variety of symptoms for the
0.1% of highest level users, about 470 people.
////////////////////////////////////////////////////////////


Naturally, I want to cut to the chase with pertinent critical comments,
often giving quotes from the ASE text and its 415 references:

"As aspartame is completely hydrolyzed following intake, studies
employing either intraperitoneal administration or direct exposure of
cells in vitro to intact aspartame do not reflect human exposures and
therefore, must be carefully interpreted." [ Spot on! ]


3.1.1 [ 22 mg ingested aspartame releases 2.4 mg methanol, which is 11%
of the aspartame. Stegink, 1987

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). If
30% of the methanol is turned into formaldehyde, the amount of
formaldehyde, 37 mg, is 18 times the USA EPA limit for daily
formaldehyde in drinking water, 2 mg in 2 L water.

For instance, hangover researchers claim that it is the ~150 mg/L
methanol impurity, about one part in 10,000, twice the level from
aspartame in diet sodas, in dark wines and liquors that, turned into
formaldehyde and then formic acid, is the major cause of the dreadful
symptoms of "morning after" hangover' ]


[reference 254:
J. Nutrition 1973 Oct; 103(10): 1454-1459.
Metabolism of aspartame in monkeys.
Oppermann JA, Muldoon E, Ranney RE.
Dept. of Biochemistry, Searle Laboratories,
Division of G.D. Searle and Co. Box 5110, Chicago, IL 60680

They found that about 70% of the radioactive methanol in aspartame put
into the stomachs of 3 to 7 kg monkeys was eliminated within 8 hours,
with little additional elimination, as carbon dioxide in exhaled air and
as water in the urine.

They did not mention that this meant that about 30% of the methanol must
transform into formaldehyde and then into formic acid, both of which
must remain as toxic products in all parts of the body.

They did not report any studies on the distribution of radioactivity in
body tissues, except that blood plasma proteins after 4 days held 4% of
the initial methanol.

This study did not monitor long-term use of aspartame. ]


3.1.5 " Ilback et al. (2003) surveyed 1120 diabetics and reported that
average daily intakes of aspartame by children and adult diabetics were
below the ADI, for both the top 5% and 10% consumers.

Worst-case intakes were calculated for the 10 diabetic children
[ 0.1 % of adults and children ] consuming the highest amounts of
artificially sweetened foods and using the highest concentrations of
sweetener allowed in food products.

The worst-case calculation was based on the following assumptions:
(1) All sweeteners found in the diet were replaced by aspartame;
(2) the level of aspartame in foods was the maximum allowed by
regulations, not the concentration actually used in the foods; and
(3) the intake was based on the highest reported consumption of 10
diabetic children consuming the highest amounts of artificially
sweetened foods.

These worst-case calculations showed that consumption by this small
subpopulation would exceed the ADI by only a small percentage (about
114% of the ADI), and thus the authors concluded that there is a
sufficient safety margin for aspartame, even in high consuming diabetics
in a single time point (Ilback et al., 2003). "

"Therefore, a short period of exposure slightly above the ADI is not
considered to pose any significant risk (Larsen and Richold, 1999)."

3.5 "Methanol generated from aspartame is estimated to average 33 mg/day
or 0.55 mg/kg bw/day for a 60-kg individual.

For individuals in the 95th percentile the estimated methanol
consumption figures are 93.9 mg/day or 1.57 mg/kg bw/day for a 60-kg
individual." [ three times more than the average ]

[ Well, 114% of the European ADI of 40 mg/kg bw/day for a 60-kg teenager
gives 2,736 mg aspartame daily, with the 11% methanol 301 mg daily, so
30% retention of cumulative durable toxic products of formaldehyde and
formic acid would be 90 mg daily -- 2700 mg a month.

About 150 mg methanol impurity in ordinary liquors produces "morning
after" hangovers, due to the conversion of methanol into formaldehyde
and formic acid.

It is urgent to assess the specific health status of this "small
subpopulation" of teenage diabetics who endure these remarkable toxic
exposures.

Many aspartame reactors used over 12 12-oz cans daily diet soda for
years. At 200 mg each, this is 2400 mg daily aspartame, 264 mg
methanol, and at 30%, 80 mg retained formaldehyde and formic acid
products -- disposition, biochemistry, and concentrations unknown as yet
to world science.

This fundamental crucial ignorance is hardly grounds for ingeniously
dismissing every one of hundreds of mainstream research reports and
thousands of individual cases by informed professionals and affected
people.

Yet, this is all that is offered the ASE review, by a paid expert panel
set up by the Burbank Group, funded by Ajinomoto.

The greatest gift of science to humanity since 1600 is the vision of
unfettered, yet disciplined, curiosity, publicly and respectfully
shared, about all aspects and levels of our infinite reality.

Questions raised generate partial answers, which open up ever expanding
frontiers of more questions, within a overall richness of understanding
and subtlety of communication that is profoundly meaningful, awesomely
beautiful, and of the greatest practical value.

Like all the other previous reviews, invariably committee work at its
worst, the ASE review, fails totally at being curious, at spotting
telling details (where frisky devils cavort), acknowledging fundamental
unknowns, and raising pointed questions to inspire their peers.

Once again, good, intelligent, trained, experienced, dedicated, earnest
scientists have failed themselves and science, by acquiescing to
participation, true, very well paid, in a ceremonial public ritual dance
of groupthink to support vast vested interests.

Once again, the good tidings are immediately proclaimed in a global
media campaign, a Katrina deluge of relentless, repetitious, pious,
preaching -- Our witch doctors have met and burned much incense, the
baleful concerns are all laid to rest, so go ye villagers and consume ye
yet more in peace.

Once again, but with some progress.

Now, there are many times more references, with more on recent studies.

Now, there is explicit discussion, or at least mention, of areas of
ignorance.

Now, at least on the edge of the table of discourse, are closely related
fields of methanol, formaldehyde, formic acid, alcohol hangover, MSG,
additives, tobacco, addiction, vehicle exhaust, diet, genetic variation,
drug interactions, the many new modern diseases, whole population
epidemiology, global data bases mining -- all growing exponentially,
all intimately, instantly linked on the Net.

Now, every match sets off fire.

Now, there are Net groups with permanent, open archives, where citizens
and experts colaborate on every problem, ie opportunity.

AspartameNM-------------------- 82 members--- 1,472 posts
Aspartame------------------- 1,047 members-- 21,510 posts
GlutenFreeCaseinFreeKids--- 11,032 members- 281,148 posts

The ASE review will ignite a new level of creative world collaboration
to finally remove gratuitous toxins.

This is the purpose of this critical review.

Now, corporations are inevitably competing to maximize the health,
energy, clarity, intelligence, sanity, and creative productivity not
only of their staff, but their customers, and all people.

A huge opportunity: find a safe, inexpensive process to remove all
methanol from alcohol beverages, solving the billion-dollar disease of
alcohol hangovers.

A corporate tipping point: whether to continue the mistake of continiung
to bet the ranch on a single highly profitable, but sadly toxic product,
in a radically uncontrolled information environment that can this year
produce a Enron-level meltdown, with tedious, hundred billion dollar
civil and criminal liabilities, a la tobacco, or immediately shut down
the operation, release all secret files and research, and set up hundred
billion dollar funds to recompense individuals and governments, while
funding independent groups to definitively study the health issues --
which can rapidly lead to good business opportunities.

Corporations probably best cooperate to expedite this forward moving
retreat.

ASE members and Burdock Group: The ASE review should be made available
for free on the Net, along with the full text of the references and many
more related studies and a open, public archive, unedited public action
group for civil discussion and collaboration.

All financial and secrecy contracts should be made public, along with
the details of the history of meetings and communications, especially
how dissent and conflict were managed.

The ASE review should best evolve into a on-going public collaboration
like Wikipedia or Citizendium. ]


3.2 "Thus there is no established UL for either aspartic acid or
phenylalanine (Institute of Medicine, 2005)."


3.3 "Taucher et al. (1995) estimated that humans produce approximately
1000 mg of methanol daily from [pectins in] fruits and vegetables."

reference 375:
Alcohol Clin Exp Res. 1995 Oct; 19(5): 1147-50.
Methanol in human breath.
Taucher J, Lagg A, Hansel A, Vogel W, Lindinger W.
Institut fur Ionenphysik, Universitat Innsbruck, Austria.

Using proton transfer reaction-mass spectrometry for trace gas analysis
of the human breath, the concentrations of methanol and ethanol have
been measured for various test persons consuming alcoholic beverages and
various amounts of fruits, respectively.

The methanol concentrations increased from a natural (physiological)
level of approximately 0.4 ppm up to approximately 2 ppm a few hours
after eating about 1/2 kg of fruits,
and about the same concentration was reached after drinking of 100 ml
brandy containing 24% volume of ethanol and 0.19% volume of methanol.
[ 24 ml = 61 g ethanol, and 0.19 ml = 0.34 g = 340 mg methanol ]
PMID: 8561283 ]

[ This important, substantial result demands thorough, immediate,
definitive research into the disposition of this apparently highly toxic
dose of methanol in various vulnerable groups of people -- rather than
be misused as an insipid ploy to insinuate that any methanol from
aspartame is a trivial dose. ]


" 3.3.1. Exposure to Formaldehyde From Methanol in Aspartame

As is described later, methanol is metabolized to formaldehyde, which is
rapidly further metabolized."

"For example, the demethylation of the caffeine found in one cup of
coffee produces 30 mg of formaldehyde (Imbus, 1988)."

[ This important, surprising result demands thorough, immediate,
definitive research into the disposition of this apparently highly toxic
source of methanol in various vulnerable groups of people -- rather than
be used as an insipid ploy to insinuate that any methanol from aspartame
is a silly issue. ]


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

"The EPA recommends that an adult should not drink water containing more
than 1 milligram of formaldehyde per liter of water (1 mg/L) for a
lifetime exposure, and a child should not drink water
containing more than 10 mg/L for 1 day or 5 mg/L for 10 days."

http://groups.yahoo.com/group/aspartameNM/message/1140
EPA Preliminary Remedial Goals, PRGs, 2003 Oct, air and tap water --
methanol, formaldehyde, formic acid -- not mentioned is methanol from
aspartame, dark wines and liquors: Murray 2004.11.20

http://groups.yahoo.com/group/aspartameNM/message/1108
faults in 1999 July EPA 468-page formaldehyde profile:
Elzbieta Skrzydlewska PhD, Assc. Prof., Medical U. of Bialystok, Poland,
abstracts -- ethanol, methanol, formaldehyde, formic acid, acetaldehyde,
lipid peroxidation, green tea, aging: Murray 2004.08.08 2005.07.11 ]

"The 1999 July EPA 468-page formaldehyde profile admits that four states
substantially exceed the federal EPA limit:

Environmental Protection Agency 2.00 mg in 2 L daily drinking water
California and Maine ---------- 0.06 mg
Maryland ---------------------- 0.02 mg
New Jersey -------------------- 0.20 mg"

Maryland's limit is ten times more stringent than the EPA's." ]



[ 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@...
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


http://groups.yahoo.com/group/aspartameNM/message/1052
DMDC: Dimethyl dicarbonate 200mg/L in drinks adds methanol 98 mg/L
( becomes formaldehyde in body ): EU Scientific Committee on Foods
2001.07.12: Murray 2004.01.22

[ DMDC would be a very useful alternative source of methanol for human
studies, as controls to compare with aspartame and wines. ] ]


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@...
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.] ]



"Formic acid is ultimately converted to CO2 and water, via the formation
of 10-formyl tetrahydrofolate (Barceloux et al., 2002)."

6.9.2.1 "Formic acid accumulates in the blood because its half-life
(t1/2 = 3.4-6 h) is very much longer than is that of formaldehyde (t1/2
= 1.5 min) (Hantson et al., 2005).

Formic acid accumulation is considered the mechanism of toxicity of high
doses of methanol, which induces metabolic acidosis, ophthalmic toxicity
and central nervous system depression (Barceloux et al., 2002)."

[ In a few pages, I give many details about formaldehyde and formic acid
disositions in blood and tissues of 4 small monkeys from a single damage
level methanol dose:

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
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."

"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." ]


Toxicol Sci. 2005 Nov; 88(1): 30-8. Epub 2005 Aug 10.
Uptake and disposition of inhaled methanol vapor in humans.
Ernstgård L, Shibata E, Johanson G.
Work Environment Toxicology, Institute of Environmental Medicine,
Karolinska Institutet, Stockholm, Sweden. Lena.Ernstgard@...
http://toxsci.oxfordjournals.org/cgi/content/full/88/1/30
free full text

Methanol is a widely used solvent and a potential fuel for motor vehicles.

Human kinetic data of methanol are sparse.

As a basis for biological exposure monitoring and risk assessment, we
studied the inhalation toxicokinetics of methanol vapor in four female
and four male human volunteers during light physical exercise (50 W) in
an exposure chamber.

The relative uptake of methanol was about 50% (range 47-53%).

Methanol in blood
increased from a background level
of about 20 to 116 and 244 microM/L after 2 h exposure
at 0, 100 ppm (131 mg/m3), and 200 ppm (262 mg/m3), respectively.

Saliva showed substantially higher levels than blood immediately after
exposure.

This difference disappeared in a few minutes;
thereafter the concentrations and time courses
in blood, urine, and saliva were similar,
with half times of 1.4, 1.7, and 1.3 h, respectively.

The postexposure decrease of methanol in exhaled air was faster,
with a half time of 0.8 h.

The methanol concentrations were approximately twice as high
in all four types of biological samples at 200 compared to 100 ppm.

No increase in urinary formic acid was seen in exposed subjects.

Our study indicates non-saturated, dose-proportional kinetics of
methanol up to 200 ppm for 2 h.

No gender differences were detected.

Similar, parallel patterns were seen with regard to the methanol time
courses in blood, urine, and saliva, whereas the concentration in
exhaled air decreased markedly faster.

Thus, apart from blood and urine, saliva also seems suitable for
biomonitoring of methanol exposure. PMID: 16093526


"Symptom ratings.
The subjects rated the level of perceived discomfort immediately before,
during (10, 50, 80, and 104 min), and after (126 and 210 min) each
exposure session.

Ten questions were answered,
related to irritative symptoms (eyes, nose, and throat or airways),
the central nervous system (headache, fatigue, nausea, dizziness,
feeling of intoxication),
difficulty in breathing,
and smell of solvent.

The ratings were performed using a 100-mm visual analogue scale
(Kjellberg et al., 1988Go)
graded from "not at all" (corresponding to 0 mm) through "hardly,"
"slightly," "fairly," "much," to "almost unbearable" (100 mm).

The same questionnaire has been used in several chamber inhalation
studies performed with organic solvent vapors in our laboratory (see for
example, Ernstgård et al., 1999Go, 2002Go; Järnberg et al., 1996Go;
Nihlén et al., 1998bGo)."

"Background levels of methanol were detected in all samples during
control exposure:
blood range 9–76 µM/L, saliva 4–76 µM/L, urine 13–86 µM/L, and exhaled
air 0.0005–0.01 µM/L.

For each individual and time point, the exposure-related methanol
concentrations were calculated as the difference between the
concentrations measured at methanol exposure
and that measured at clean air exposure.

No difference between genders was seen with respect to background
methanol in blood.

Background methanol levels in urine were higher in men than in women
(35.9 vs. 21.5 µM/L, p = 0.03 in t-test).

Similar difference was seen for saliva (39.3 vs. 19.0 µM/L, p = 0.008).

Methanol was rapidly absorbed by inhalation.

The relative uptake remained stable throughout the exposure and was
approximately 50% at both exposure levels (range 47–53%) (Table 1).

The blood methanol concentrations reached 116 (94–144) µM/L after 2 h
exposure at 100 ppm and 244 (228–260) µM/L at 200 ppm methanol.

These levels are consistent with linear, nonsaturated metabolism of
methanol.

Linear (i.e., dose-proportional) kinetics is also indicated when
comparing the AUC (0–6 h) of blood methanol at the different exposure
levels (Fig. 1A).

Blood methanol increased in a monoexponential fashion during exposure
(Fig. 2A).

The postexposure decline was also monoexponential, considering the
background methanol.

According to the toxicokinetic model, the elimination half time in blood
was about 1.4 h, and the apparent total clearance 0.2–0.3 l/min.

The steady-state level of methanol at continuous exposure to methanol
was calculated to be 186 and 394 µM at 100 and 200 ppm, respectively
(Table 1), again an indication of linear kinetics."


reference 133:
Bull Mem Acad R Med Belg. 2006;161(6):425-34; discussion 434-6.Links
[Acute methanol intoxication: physiopathology, prognosis and treatment]
[Article in French]
Hantson PE.
Département des Soins Intensifs, Cliniques St-Luc-U.C.L.

Acute methanol poisoning is mainly the consequence of voluntary or
accidental ingestion.

The mortality and morbidity rates remain very high despite intensive
care therapy.

Methanol by itself is poorly toxic.

Methanol is transformed in the liver into formaldehyde and thereafter
formic acid

Metabolic acidosis is the main biological feature of poisoning.

Acidosis is related to formic acid accumulation, and also to a less
extent to lactate production.

In contrast to rodents, primates are relatively deficient in
tetrahydrofolate reductase and therefore formic acid is usually the
final metabolite.

Formic acid is able to inhibit cytochrome oxidase activity in the
mitochondria, leading to histotoxic hypoxia.

The most sensitive organs to the effects of formic acid are the brain
and the visual pathway, while other organs may also be seriously damaged
according to the severity of metabolic acidosis.

Hemodialysis remains indicated for the removal of both methanol and
formic acid.

Fomepizole is a recently approved antidote. It appears safe and effective.

Analysis of its cost-effectiveness ratio is still ongoing in methanol
poisoning. PMID: 17288275


Am J Emerg Med. 2006 Oct; 24(6): 725-8.
Inhalational abuse of methanol products: elevated methanol and formate
levels without vision loss.
Bebarta VS, Heard K, Dart RC.
Department of Emergency Medicine, Wilford Hall Medical Center, San
Antonio, TX, USA. vikbebarta@...

Inhalant abuse of methanol-containing products has increased over the
last decade.

We performed a prospective observational study of 7 subjects who
presented to an ED after inhalant abuse of methanol-containing
hydrocarbon products.

Four patients had a methanol level greater than 24 mg/dL and 2 had an
anion gap greater than 17 mEq/L. [ 240 mg/L blood ]

The mean formic acid level was 71 microg/mL, and 1 patient had a level
considered high enough to induce retinal toxicity (>200 microg/mL).
[ 71 mg/L blood]

No patient had an abnormal ophthalmologic examination.

All patients were treated with intravenous folate,
2 received alcohol dehydrogenase blockade,
and no patient received hemodialysis or intravenous bicarbonate.

All patients' acidosis resolved within 4 hours.

The methanol and formic acid levels are lower than those reported after
methanol ingestion.

These preliminary data suggest that inhalant abusers of methanol
products may have significantly elevated methanol and formic acid
levels, but are at low risk for methanol induced complications of visual
dysfunction and refractory acidosis. PMID: 16984844


J Anal Toxicol. 2005 Sep; 29(6): 586-8.
Increased serum formate in the diagnosis of methanol poisoning.
Hovda KE, Urdal P, Jacobsen D.
Department of Acute Medicine, Ullevaal University Hospital,
NO-0407 Oslo, Norway. knov@...

Early diagnosis is essential for successful treatment in methanol poisoning.

Methanol detection by gas chromatography is not available in most hospitals.

Methanol increases the osmolal gap in serum and its metabolite formate
increases the anion gap.

The sensitivity of these indirect diagnostic methods is not good at low
concentrations of methanol or formate.

We therefore studied the usefulness of formate measurement in diagnosing
methanol poisoning.

In 15 patients poisoned with methanol, serum formate was measured
enzymatically on a Cobas Mira analyzer using formate dehydrogenase and
nicotinamid adenine dinucleotid.

Day-to-day coefficient of variation was 5%, and the upper reference
limit was 2 mg/dL (0.4 mmol/L). [ 20 mg/L blood serum = 0.4 mmol/L ]

Methanol was detected in all 15 patients of whom 14 had elevated serum
formate concentrations.

Anion gap was increased in 11 of 11, and osmolal gap in 11 patients of
15 examined.

Metabolic acidosis was present in 12 of 15 patients, but pH was below
7.30 in only 9 of them.

Four patients with no symptoms had formate concentrations in the range
2-38 mg/dL (0.5-8.3 mmol/L), indicating that increased serum formate was
a sensitive indicator of methanol poisoning.
[ 20-380 mg/L blood serum = 0.5-8.3 mmol/L]

Our results proved formate analyzes to be a simple, sensitive, and
specific way of diagnosing methanol poisoning.

Confounders are patients admitted early, or concomitant ethanol
ingestion, and therefore no acidosis.

This problem may, however, be omitted by repeated formate analysis in
patients developing metabolic acidosis. PMID: 16168185


[ It is remarkable how little is known about the disposition of
formaldehyde and formic acid in human tissues, according to a sober
review by Bouchard M, 2001:

"Exposure to methanol also results from the consumption of certain
foodstuffs (fruits, fruit juices, certain vegetables, aspartame
sweetener, roasted coffee, honey) and alcoholic beverages (Health
Effects Institute, 1987; Jacobsen et al., 1988)."

"A biologically based dynamic model was developed to simulate the uptake
and disposition of methanol and its metabolites (formaldehyde, formate,
CO2) in animals and humans."

"Systemic methanol is extensively metabolized by liver alcohol
dehydrogenase and catalase-peroxidase enzymes to formaldehyde, which is
in turn rapidly oxidized to formic acid by formaldehyde dehydrogenase
enzymes (Goodman and Tephly, 1968; Heck et al., 1983; Røe, 1982; Tephly
and McMartin, 1984)."

"Formaldehyde, as it is highly reactive, forms relatively stable adducts
with cellular constituents (Heck et al., 1983; Røe, 1982)."

"Thus, in monkeys and plausibly humans, a much larger fraction of body
formaldehyde is rapidly converted to unobserved forms rather than passed
on to formate and eventually CO2."

"Inversely, in monkeys and in humans, a larger fraction of body burden
of formaldehyde is rapidly transferred to a long-term component.
The latter represents the formaldehyde that (directly or after oxidation
to formate) binds to various endogenous molecules..."

"However, fits to the available data in rats and monkeys of Horton et
al. 1992) and Dorman et al. (1994) show that, once formed, a substantial
fraction of formaldehyde is converted to unobserved forms.

This pathway contributes to a long-term unobserved compartment.

The latter, most plausibly, represents either the formaldehyde that
(directly or after oxidation to formate) binds to various endogenous
molecules (Heck et al., 1983; Røe, 1982)...

That substantial amounts of methanol metabolites or by-products are
retained for a long time is verified by Horton et al. (1992) who
estimated that 18 h following an iv injection of 100 mg/kg of
14C-methanol in male Fischer-344 rats, only 57% of the dose was
eliminated from the body.

From the data of Dorman et al. (1994) and Medinsky et al. (1997), it can
further be calculated that 48 h following the start of a 2-h inhalation
exposure to 900 ppm of 14C-methanol vapors in female cynomolgus monkeys,
only 23% of the absorbed 14C-methanol was eliminated from the body.

These findings are corroborated by the data of Heck et al. (1983)
showing that 40% of a 14C-formaldehyde inhalation dose remained in the
body 70 h postexposure."

http://www.toxsci.oupjournals.org/cgi/content/full/64/2/169
free full text
Toxicological Sciences 64, 169-184 (2001)
A Biologically Based Dynamic Model for Predicting the Disposition of
Methanol and Its Metabolites in Animals and Humans
Michèle Bouchard *, #,1, bouchmic@...
Robert C. Brunet, # brunet@...
Pierre-Olivier Droz, #
Gaétan Carrier* gaetan.carrier@...

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 2005.01.05 ]



" ACKNOWLEDGMENTS

This safety review was conducted by the Burdock Group, at the request of
the sponsor.

Dr. William Waddell was selected as the chair of the review expert
panel, and had a free hand in selection of the panelists.

As described in the introduction, panelists were chosen to achieve
representation of the complete spectrum of toxicological expertise
relevant to aspartame

The identity of the sponsor of the review was unknown to the chair and
the expert panelists throughout the conduct and completion of the review.

The identity of panelists also remained unknown to the sponsor.

The Burdock Group managed reimbursement of panelists.

There were no known conflicts of interest or potential biases of the
authors.

This review represents the professional views of the authors.

The authors dedicate this article to Dr. Robert M. Kroes, who actively
participated in the preparation of this paper and approved the final
version before he died from rapidly progressing lung cancer. "

[ I did a quick survey on the distinguished panel, which shows an
obvious predominant pro-industry bias, in terms of close association
with aspartame firms, their experts and their groups:

BA Magnuson works for the Burdock Group, while GA Burdock wrote a text
chapter with Frank Krukonis, a NutraSweet Co. VP.

Clearly, GA Burdock's career is entirely about serving industry in its
dealings with government health regulation.

J Doull took the industry side re mercury amalgam safety in 2005.

GM Marsh published 9 large formaldehyde studies, which guarantees his
expertise, but runs the risk that an expert may be locked into his
established opinions.

R Walker expressed unqualified approval of aspartame in 1999.

Certainly, no one was selected who already had an unbiased,
sophisticated command of the vast literature of the aspartame controversy.

Many central, highly scientific reviews and studies are not in the 415
references:

http://groups.yahoo.com/group/aspartameNM/message/870
Aspartame: Methanol and the Public Interest 1984: Monte:
Murray 2002.09.23 rmforall

Dr. Woodrow C. Monte Aspartame: methanol, and the public health.
Journal of Applied Nutrition 1984; 36 (1): 42-54.
(62 references) Professsor of Food Science [retired 1992]
Arizona State University, Tempe, Arizona 85287


http://groups.yahoo.com/group/aspartameNM/message/1067
eyelid contact dermatitis by formaldehyde from aspartame,
AM Hill & DV Belsito, Nov 2003: Murray 2004.03.30 [150 KB]

[ In this 2003 critique, I added a long critique of industry research,
including this study, not in the ASE review:

Biochemical Pharmcacology 1979: 28; 645-649. [ No abstract in PubMed ]
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
The Toxicology Center, Dept. of Pharmacology,
University of Iowa, Iowa City, Iowa 52242 PMID: 109089
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 the 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]


Appendix F:

The exponential fragmentation of science into a fractile structure of
ever more atomized specialties ensures that every expert is a layman
outside his own specialty.

Capable laymen play an essential role by summarizing and integrating
scattered lines of inquiry that certain vested interests have long-term
campaigns for obscuring, since outright opposition would tend to attract
discussion and scrutiny that would soon vitiate billion dollar products.
Most professionals simply do not have the free time to investigate such
arcane, but possibly crucial, details. Capable laymen now join together
on the Net to establish credibility by common sense, polite mobilization
of specialized research, backed by support from informed specialists.

For instance, I started investigating aspartame in early January 1999
and within two months was being given papers by Woodrow C. Monte and
Ralph G.Walton.

The route of aspartame to methanol to formaldehyde to formic acid is a
classic example. Were this line of inquiry already suspected to be sure
to establish the harmlessness of aspartame, then the industry would have
every motive to spend a few paltry millions to both complete the
research in humans and widely publicize the results.

The fact that on the contrary, there is no industry funded research in
humans at all in the public domain on the specific biochemical and
tissue outcomes of formaldehyde and formic acid from aspartame leads to
a reasonable surmise that the industry has reason to fear, obscure, and
derail this inquiry. Following the crooked but unmistakable trail of
missing research, i.e., avoided, ignored, misstated, discounted,
obscured, explained away, or simply never mentioned, is an excellent
strategy for uncovering the lurking secret.

In spring 1999, an eminent pro-aspartame scientist Christian Tschanz had
NutraSweet Co. give me their $ 130 review text of their research, "The
Clinical Evaluation of a Food Additive: Assessment of Aspartame" (1996),
by Christian Tschanz, Harriett H. Butchko, W. Wayne Stargel, and Frank
N. Kotsonis, all apartame stalwarts.

Chapter 5: "Metabolism and Pharmacokinetics of Radiolabeled Aspartame in
Normal Subjects", by Aziz Karim and Thomas Burns, has 10 pages and 10
citations. Page 63, Figure 4, Metabolic products derived from aspartame,
beta-aspartame, and DKP, does not list formaldehyde or formic acid.

The tangle of black arrows includes two paths from Aspartame to Methanol
to "CO2 + Body Constituents". Now, that's pretty good public relations
spin, eh?

"Body Constituents", indeed? This is systematic and persistent deceit,
as pernicious as it is profitable.

Aziz Karim, PhD is a "Distinguished Research Fellow and Sr. Director,
Clinical Research, G.D. Searle and Company, Skokie, Illinois", where
Thomas Burns, M.S. is a "Clinical Research Manager".

They state that "in monkeys" with methanol or aspartame labelled in the
methyl ester, both with 14C, "...excretion of 14CO2 in the expired air
occured to the same extent (about 70% of the 14C dose) with both
compounds, indicating complete hydrolysis of the methyl ester moiety of
aspartame (Figure 6)."

They said nothing about resulting levels in blood plasma,
urine, feces, or any body tissues.

This is the typical commission by omission strategy
of industry research on aspartame.

reference 254:
J. Nutrition 1973 Oct; 103(10): 1454-1459.
Metabolism of aspartame in monkeys. [ No abstract in PubMed ]
Oppermann JA, Muldoon E, Ranney RE.
Dept. of Biochemistry, Searle Laboratories,
Division of G.D. Searle and Co. Box 5110, Chicago, IL 60680
http://jn.nutrition.org/cgi/reprint/103/10/1454 free full text

They found that 68.67% of the radioactive methanol in aspartame put into
the stomachs of 3 to 7 kg monkeys was eliminated within 8 hours, with
little additional elimination afterwards, as carbon dioxide in exhaled
air and in the urine.

They did not mention that this meant that about 31.33% of the methanol
must transform into formaldehyde and then into formic acid, both of
which must remain as toxic products in all parts of the body.

They did not report any studies on the distribution of radioactivity in
body tissues, nor give the absolute levels for declining blood plasma
proteins.

This study did not monitor long-term use of aspartame, which might
reveal cumulative effects.

Their low oral dose of aspartame and for methanol was 0.068 mmol/kg,
about 1 part per million [ppm] of the acute toxicity level of 2,000
mg/kg, 67,000 mmol/kg, used by McMartin (1979).

Two L daily use of diet soda provides 123 mg methanol, 2 mg/kg for a 60
kg person, a dose of 63 mmole/kg, a thousand times more than the dose in
this study.

By eight hours excretion of the dose in air and urine had leveled off at
67.1 +-2.1% as CO2 in the exhaled air and 1.57+-0.32% in the urine, so
68.7% was excreted, and 31.3% was retained. [This data is the average of
4 monkeys.]
"...the 14C in the feces was negligible."
"That fraction not so excreted (about 31%) was converted to body
constituents through the one-carbon metabolic pool."

"All radioactivity measurements were counted to +-1% accuracy..."
This indicates that the results could not be claimed to have a precision
of a tenth of a percent. OK, so this is a nit-pick-- but I believe
espousing spurious accuracy is a sign of scientific insecurity.

The abstract ends, "It was concluded that aspartame was digested to its
three constituents that were then absorbed as natural constituents of
the diet."
Thus, the concept is very subtly insinuated that methanol, as a
constituent of aspartame, is absorbed as a natural constituent of the
diet. "Dietary methanol is derived in large part from fresh fruits and
vetetables."
Nowhere in this report, or in the book chapter are mentioned the dread
words, "formaldehyde" and "formic acid".


Searle Laboratories team in 1976 reported that in 4 monkeys
fed aspartame, by 12 hours: "...the major fraction (70 %)
of the [aspartate] label appeared in the expired air (Fig.6)...
Urinary and fecal 14C [ aspartate derived ]
amounted to 4--6 % of the administered [ aspartate ] label."

This gives a total of a maximum 76 % excreted aspartate
from the aspartame, indicating that 24 % of this excitotoxin
was retained in the body. It is reasonable to conclude
that daily use of aspartame must lead to substantial
accumulation of this excitotoxin, aspartate, in body tissues.

Their 1979 review said: "Aspartame... is hydrolyzed in the gut
to yield aspartic acid, phenylalanine, and methanol....
Aspartate may also be incorporated into body constitutents
such as other amino acids, proteins, pyrimidines, asparagine,
and N-acetylaspartic acid."

276: J Environ Pathol Toxicol. 1979 Mar-Apr; 2(4): 979-85.
A review of the metabolism of the aspartyl moiety of aspartame in
experimental animals and man.
Ranney RE, Oppermann JA.
Department of Drug Metabolism and Radiochemistry,
Searle Laboratories, Skokie, Illinois.
Division of G.D. Searle and Co. Box 5110, Chicago, IL 60680

Aspartame (3-amino-N-(alpha-carboxyphenethyl) succinamic acid,
methyl ester; the methyl ester of aspartylphenylalanine, SC-18862) is
hydrolyzed in the gut to yield aspartic acid, phenylalanine, and methanol.
This review of the literature describes the metabolic paths
followed by aspartate in its conversion to CO2
or its incorporation into body constituents.

About 70 percent of 14C from [asp-14C]-aspartame is converted
in the monkey to 14CO2.

Some of the aspartate is converted at the intestinal mucosal level to
alanine by decarboxylation.
This amino acid may be oxidized to CO2 by entering
the tricarboxylic acid cycle via pyruvate and acetyl CoA.
In addition, transamination of aspartate to oxaloacetate permits
this product also to enter the tricarboxylic acid cycle.

Aspartate may also be incorporated into body constitutents
such as other amino acids, proteins, pyrimidines, asparagine,
and N-acetylaspartic acid.

It is concluded that the aspartate moiety of aspartame
is metabolized in a manner similar to that of dietary aspartic acid.
Publication Types: Review PMID: 376770


J. A. Oppermann et al, plus F.G. McMahon of Tulane University Medical
School, published a follow-up study, "Comparative metabolism of
aspartame in experimental animals and humans", J. Toxicology and
Environmental Health 2: 441-451, 1976.

The abstract says, "Hydrolysis of the methyl group by intestinal
esterases yielded methanol, which was oxidized in the one-carbon
metabolic pool to CO2."

"The hypothetical pathways of metabolism, which aspartame was expected
to follow, are diagrammed in Fig. 1....The principle used to test the
validity of this hypothetical description of the metabolism of aspartame..."

Figure 1. shows in an nice orderly sequence that:
(a) MeOH ---> one-carbon metabolic pool ---> CO2 + formyl metabolites .

Meanwhile, this sentence jumps from p. 441 to 442 under Figure 1., "The
absorbed methanol would be incorporated into the one-carbon pool and
would be converted [ page jump in sentence ] primarily to CO2 (Makar
et.al., 1968; Tephly et al, 1964), although a small fraction might be
incorporated into body constituents."

The graphs present the same methanol in monkey data as in 1973, but the
nowhere is the specific percentage of exhaled CO2 mentioned.

Methanol and aspartame were also given to a few [ unspecified ] number
of rats: "The major fraction of the 14C was excreted in the expired air
(Fig. 2)...Plasma levels of 14C reached a peak
[ absolute data not given ] at about 3 hr..."

In this follow-up report, for methanol and the methyl group in
aspartame, excretion in urine and feces were not mentioned in either the
former monkey or the new rat studies, the absolute plasma levels were
not given, and, of course, no measures were taken of 14C in body tissues.

The only hint of the possible role of formaldehyde and formic acid was
the rather diffident term "formyl metabolites" in Figure 1.

Overall, we see consistent patterns of avoiding any focus on the actual
disposition of extremely toxic formaldehyde and formic acid, both
persistent and cumulative, products in body tissues.

Subtle equivocation and qualification was expressed by such words as
"hypothetical", "was expected to follow", "would be", "primarily",
"although a small fraction might be incorporated into body
constituents", "major fraction".

Methanol from aspartame was not studied in the other species: rabbits,
dogs, and humans.

It pays to investigate early studies, because then the coverup was less
well organized, more patchy.

The loosely organized world-wide exponential growth of science ensures
that the line of inquiry of methanol to formaldehyde and formic acid
will pop up here and there, but no one is encouraged to make the
connection with aspartame, widely proclaimed as "the most thoroughly
tested food additive in history" -- until the momentous, unheralded
Trocho study established explosive results in June 1998. (Appendix E)


C. Trocho (1998):
"In all, the rats retained, 6 hours after administration, about 5 % of
the label, half of it in the liver."

They used a very low level of aspartame ingestion, 10 mg/kg, for rats,
which have a much greater tolerance for aspartame than humans.

So, the corresponding level for humans would be about 1 or 2 mg/kg.

Many headache studies in humans used doses of about 30 mg/kg daily.

http://groups.yahoo.com/group/aspartameNM/message/925
aspartame puts formaldehyde adducts into tissues, Part 1/2
full text, Trocho & Alemany 1998.06.26: Murray 2002.12.22

http://ww.presidiotex.com/barcelona/index.html full text
Formaldehyde derived from dietary aspartame
binds to tissue components in vivo.
Life Sci June 26 1998; 63(5): 337-49.
Departament de Bioquimica i Biologia Molecular,
Facultat de Biologia, Universitat de Barcelona, Spain.
http://www.bq.ub.es/cindex.html Línies de Recerca: Toxicitat de
l'aspartame http://www.bq.ub.es/grupno/grup-no.html
Sra. Carme Trocho, Sra. Rosario Pardo, Dra. Immaculada Rafecas,
Sr. Jordi Virgili, Dr. Xavier Remesar, Dr. Jose Antonio
Fernandez-Lopez, Dr. Marià Alemany [male]
Fac. Biologia Tel.: (93)4021521, FAX: (93)4021559
Sra. Carme Trocho "Trok-ho" Fac. Biologia Tel.: (93)4021544,
FAX: (93)4021559 alemany@... ; bioq@...

Abstract:

Adult male rats were given an oral dose of 10 mg/kg aspartame,
14C-labeled in the methanol carbon.
[ 1.1 mg/kg methanol, with likely 30% retained as durable toxic
cumulative products of formaldehyde and formic acid, 0.3 mg/kg.

At timed intervals of up to 6 hours, the radioactivity in plasma
and several organs was investigated.

Most of the radioactivity found (>98 % in plasma, >75 % in liver)
was bound to protein.

Label present in liver, plasma and kidney was in the range
of 1-2 % of total radioactivity administered per g or mL,
changing little with time.

Other organs (brown and white adipose tissues, muscle, brain,
cornea and retina) contained levels of label
in the range of 1/12th to 1/10th of that of liver.

In all, the rats retained, 6 hours after administration,
about 5% of the label, half of it in the liver.

The specific radioactivity of tissue protein, RNA and DNA
was quite uniform.

The protein label was concentrated in amino acids,
different from methionine, and largely coincident
with the result of protein exposure to labeled formaldehyde.

DNA radioactivity was essentially in a single different adduct base,
different from the normal bases present in DNA.

The nature of the tissue label accumulated was, thus,
a direct consequence of formaldehyde binding to tissue structures.

The administration of labeled aspartame to a group of cirrhotic rats
resulted in comparable label retention by tissue components,
which suggests that liver function (or its defect) has little effect
on formaldehyde formation from aspartame
and binding to biological components.

The chronic treatment of a series of rats with 200 mg/kg of
non-labeled aspartame during 10 days results in the accumulation
of even more label when given the radioactive bolus,
suggesting that the amount of formaldehyde adducts
coming from aspartame in tissue proteins and nucleic acids
may be cumulative.

It is concluded that aspartame consumption may constitute
a hazard because of its contribution
to the formation of formaldehyde adducts. PMID: 9714421

[ Extracts ]
"The high label presence in plasma and liver is in agreement with the
carriage of the label from the intestine to the liver via the portal vein.

The high label levels in kidney and, to a minor extent, in brown adipose
tissue and brain are a consequence of their high blood flows (45).

Even in white adipose tissue, the levels of radioactivity found 6 hours
after oral administration were 1/25th those of liver.

Cornea and retina, both tissues known to metabolize actively methanol
(21,28) showed low levels of retained label.

In any case, the binding of methanol-derived carbon to tissue proteins
was widespread, affecting all systems,
fully reaching even sensitive targets such as the brain and retina....

The amount of label recovered in tissue components was quite high
in all the groups, but especially in the NA rats.

In them, the liver alone retained, for a long time, more than 2 % of the
methanol carbon given in a single oral dose of aspartame,
and the rest of the body stored an additional 2 % or more.

These are indeed extremely high levels for adducts of formaldehyde, a
substance responsible of chronic deleterious effects (33), that has also
been considered carcinogenic (34,47).

The repeated occurrence of claims that aspartame
produces headache and other neurological and psychological
secondary effects --
more often than not challenged by careful analysis -- (5, 9, 10, 15, 48)
may eventually find at least a partial explanation in the permanence
of the formaldehyde label,
since formaldehyde intoxication can induce similar effects (49).

The cumulative effects derived from the incorporation of label in the
chronic administration model suggests that regular intake of aspartame
may result in the progressive accumulation of formaldehyde adducts.

It may be further speculated that the formation of adducts can help to
explain the chronic effects aspartame consumption may induce on
sensitive tissues such as brain (6, 9, 19, 50).

In any case, the possible negative effects that the accumulation of
formaldehyde adducts can induce is, obviously, long-term.

The alteration of protein integrity and function may needs some time to
induce substantial effects.

The damage to nucleic acids, mainly to DNA,
may eventually induce cell death and/or mutations.

The results presented suggest that the conversion of aspartame methanol
into formaldehyde adducts in significant amounts in vivo should
to be taken into account because of the widespread utilization
of this sweetener.

Further epidemiological and long-term studies are needed to determine
the extent of the hazard that aspartame consumption poses for humans."


http://www.drthrasher.org/formaldehyde_1990.html full text
Jack Dwayne Thrasher, Alan Broughton, Roberta Madison.
Immune activation and autoantibodies in humans
with long-term inhalation exposure to formaldehyde.
Archives of Environmental Health. 1990; 45: 217-223.
"Immune activation, autoantibodies, and anti-HCHO-HSA antibodies
are associated with long-term formaldehyde inhalation."
PMID: 2400243

"Inhalation exposure to formaldehyde (HCHO)
is associated with symptoms of irritation to mucous membranes, (1,2)
chronic health problems (e.g., asthma, (2) nasopharyngeal cancer, (3)
and multiple subjective health complaints. (4,5))
Recent observations have shown that both humoral-and cell-mediated
immunologic mechanisms occur in humans with long-term HCHO exposure.

Antibodies of all isotypes to HCHO conjugated human serum albumin
(HCHO-HSA) are demonstrable in HCHO anaphylaxis, (6) hemodialysis
patients, (7) mobile home residents, (4) persons with occupational
exposures, (5,8) office workers, (9) and in persons in other
environments. (4)

In addition, changes in cell-mediated immunity include increases in
eosinophils, basophils, and T-suppressor cells following acute exposure
of patients with HCHO asthma. (10)

Moreover, individuals with multiple subjective health complaints
associated with long-term HCHO inhalation have evidence of immune
activation and the presence of autoantibodies. (4,5)

The patients in our study had symptoms and complaints related to several
organs, as described previously, (4,5,9) which were similar to symptoms
of workers with multiple chemical sensitivity,(11) cacosmia,(12) and
other chemical exposures. (13-15)

We report on the differences in humoral and cell-mediated immunity in
humans with long-term inhalation exposure to HCHO
vs. asymptomatic students (controls) who experienced short-term,
periodic exposure to the chemical."

[ http://lassesen.com/cfids/cacosmia.htm
Cacosmia (a.k.a. Multiple Chemical Sensitivity) Details:
* Chemical odour intolerance induced headache, itching eyes, irritated
or congested nose, dry and/or sore throat, cough, dizziness, and itching
or rash.
* Cacosmics reported increased prevalence of physician-diagnosed nasal
allergies, breast cysts, hypothyroidism, sinusitis, food sensitivities,
irritable bowel, and migraine headache. Resource: http://www.mcsrr.org ]

"Symptoms.
All patients in this study had sought continuous medical
attention because of multiple organ symptoms involving the central
nervous system (CNS) (headaches, memory loss, difficulty completing
tasks, dizziness), upper- and lower-respiratory symptoms,
skeletal-muscle complaints, and gastroenteritis.

Three common symptoms were expressed:
[1.] and initial flu-like illness from which they had not fully
recovered;
[2.] chronic fatigue; and
[3.] an olfactory sensitivity to ambient conditions containing low
concentrations of chemicals. (4,9,11)"

"It is recognized that chemicals and therapeutic drugs are associated
with a Lupus-like syndrome. (28,29 )
The observations made on the patients in this study support this concept."

"Five groups of subjects exposed to HCHO,
who gave informed consent, were included in this study.
[1.] Controls consisted of students of chiropractic medicine
(16 males, 12 females), mean age = 29 +- 9 y) exposed to HCHO
for 13 h/wk for 28 wk while studying human anatomy.
Immunologic tests were performed 12 mo following the last classroom
exposure.
No measurements of HCHO concentrations were made.

It is assumed that classroom ambient concentrations were at least
0.43 ppm. (1)

The students stated that during exposure they experienced
eye, nose and throat irritation and that there was a pungent odor of
HCHO.

They did not have residual health complaints (symptoms), and
they were asymptomatic at the time blood was taken.

[2.] Mobile home residents consisted of 19 patients (6 males, 13
females), mean age 41+-20 y) who currently lived in mobile homes.
The patients had lived in their environments for 2-7 y and reported
multiple symptoms. (4,9)
Measured HCHO concentrations ranged from 0.05 to 0.5 ppm at the time
blood samples were taken.

[3.] Office workers included 21 patients
(5 males, 16 females, mean age of 40 +-10 y)
who worked in new office buildings where there was inadequate
ventilation (closed buildings).
The patients had multiple health complaints. (9)

It was determined from medical histories that their symptoms commenced
with employment, waned when away from work (i.e., weekends, holidays,
vacations) and became worse upon return to work.

No HCO measurements were done; however, closed buildings have ambient
concentrations ranging from 0.01 to 0.77 ppm. (1,16)

[4.] This group included 21 patients (10 males, 11 females,
mean age of 35 + -17 y) who had multiple symptoms and who had been
removed from their original sources of HCHO exposure (mobile homes
and/or particleboard subflooring) for at least 1 y.

The HCHO concentrations measured during their exposures ranged from 0.14
to 0.81 ppm.

[5.] Ocupationally exposed patients
(6 males, 2 females, mean age of 45 + -11 y)
had HCHO exposures from the following: biology and human
anatomy classes, mortuary, pathology, physical therapy, formica
furniture (particleboard), and carbonless copy paper.
Information on six of these patients was previously published. (5)"

"In conclusion, measurements of changes in WBCs, T cells, and H/S
ratios in individuals with apparent chemical sensitivities appear to be
inadequate immune parameters to examine.

If one assumes that these individuals respond immunogically to
environmental chemicals,
investigations into autoimmunity and immune activation and
perturbations in the interleukins, luekotreines, prostglandins, and
other immunologic mediators appear to be fruitful areas for further
research. (29-32)

Thus, it appears that HCHO sensitivity is a real
phenomenon and requires further research. (4,27-32 )" ]


6.5 "Therefore, aspartame is considered to have no reproductive or
teratogenic activity, and no effect on lactation.

In these studies, effects have been observed at exceedingly high doses,
and were secondary to reduced body weights."

[ Arch Environ Health 2001 Jul-Aug; 56(4): 300-11.
Embryo toxicity and teratogenicity of formaldehyde. [100 references]
Thrasher JD, Kilburn KH. toxicologist1@...
Sam-1 Trust, Alto, New Mexico, USA.
(702) 987-4590 (505) 937-1150
http://www.drthrasher.org/formaldehyde_embryo_toxicity.html full text

"The major difference is that the Japanese
demonstrated the incorporation of FA and its metabolites
into the placenta and fetus.
The quantity of radioactivity remaining in maternal and fetal tissues
at 48 hours was 26.9 % of the administered dose." [ Ref. 14-16 ]

The DNA fraction contained 20 % and 50% of total incorporated
radioactivity in the maternal and fetal liver at 6 and 24 hours when
compared to the acid insoluble fraction (Fig. 1).

Of primary interest is that the incorporated radioactivity persisted
longer in the fetal liver and brain when compared to the mothers."

Abstract:

C-14 [radioactive labelled] formaldehyde crosses the placenta and
enters fetal tissues.
The incorporated radioactivity is higher in fetal
organs (i.e., brain and liver) than in maternal tissues.
The incorporation mechanism has not been studied fully, but formaldehyde
enters the single-carbon cycle and is incorporated as a methyl group
into nucleic acids and proteins.

Also, formaldehyde reacts chemically with organic compounds (e.g.,
deoxyribonucleic acid, nucleosides, nucleotides, proteins, amino acids)
by addition and condensation reactions, thus forming adducts and
deoxyribonucleic acid-protein crosslinks.

The following questions must be addressed:
What adducts (e.g., N-methyl amino acids) are formed in the blood
following formaldehyde inhalation? What role do N-methyl-amino adducts
play in alkylation of nuclear and mitochondrial deoxyribonucleic acid,
as well as mitochondrial peroxidation?

The fact that the free formaldehyde pool in blood is not affected
following exposure to the chemical does not mean that formaldehyde is
not involved in altering cell and deoxyribonucleic acid characteristics
beyond the nasal cavity.

The teratogenic effect of formaldehyde in the English literature has
been sought, beginning on the 6th day of pregnancy (i.e., rodents)
(Saillenfait AM, et al. Food Chem Toxicol 1989, pp 545-48;
Martin WJ. Reprod Toxicol 1990, pp 237-39;
Ulsamer AG, et al. Hazard Assessment of Chemicals; Academic Press,
1984, pp 337-400;
and U.S. Department of Health and Human Services. Toxicological Profile
of Formaldehyde; ATSDR, 1999
[references 1-4, respectively, herein]).

The exposure regimen is critical and may account for the differences in
outcomes. Pregnant rats were exposed
(a) prior to mating,
(b) during mating,
(c) or during the entire gestation period.
These regimens
(a) increased embryo mortality;
(b) increased fetal anomalies (i.e., cryptochordism and aberrant
ossification centers);
(c) decreased concentrations of ascorbic acid; and
(d) caused abnormalities in enzymes of mitochondria, lysosomes, and the
endoplasmic reticulum.

The alterations in enzymatic activity persisted 4 mo following birth.

In addition, formaldehyde caused metabolic acidosis,
which was augmented by iron deficiency.
Furthermore, newborns exposed to formaldehyde in
utero had abnormal performances in open-field tests.

Disparities in teratogenic effects of toxic chemicals are not unusual.
For example, chlorpyrifos has not produced teratogenic effects in rats
when mothers are exposed on days 6-15
(Katakura Y, et al. Br J Ind Med 1993, pp 176-82
[reference 5 herein]) of gestation
(Breslin WJ, et al. Fund Appl Toxicol 1996, pp 119-30;
and Hanley TR, et al. Toxicol Sci 2000, pp 100-08
[references 6 and 7, respectively, herein]).

However, either changing the endpoints for measurement or exposing
neonates during periods of neurogenesis (days 1-14 following birth) and
during subsequent developmental periods produced adverse effects. These
effects included neuroapoptosis, decreased deoxyribonucleic acid and
ribonucleic acid synthesis, abnormalities in adenylyl cyclase cascade,
and neurobehavioral effects
(Johnson DE, et al. Brain Res Bull 1998, pp 143-47;
Lassiter TL, et al. Toxicol Sci 1999, pp 92-100;
Chakraborti TK, et al. Pharmacol Biochem Behav 1993, pp 219-24;
Whitney KD, et al. Toxicol Appl Pharm 1995, pp 53-62;
Chanda SM, et al. Pharmacol Biochem Behav 1996, pp 771-76;
Dam K, et al. Devel Brain Res 1998, pp 39-45;
Campbell CG, et al. Brain Res Bull 1997, pp 179-89;
and Xong X, et al. Toxicol Appl Pharm 1997, pp 158-74
[references 8-15, respectively, herein]).

Furthermore, the terata caused by thalidomide
is a graphic human example in which the animal model and timing of
exposure were key factors
(Parman T, et al. Natl Med 1999, pp 582-85;
and Brenner CA, et al. Mol Human Repro 1998, pp 887-92
[references 16 and 17, respectively, herein]).

Thus, it appears that more sensitive endpoints (e.g., enzyme activity,
generation of reactive oxygen species, timing of exposure) for the
measurement of toxic effects of environmental agents on embryos,
fetuses, and neonates are more coherent than are gross terata
observations.

The perinatal period from the end of organogenesis to the
end of the neonatal period in humans approximates the 28th day of
gestation to 4 wk postpartum. Therefore, researchers must investigate
similar stages of development
(e.g., neurogenesis occurs in the 3rd trimester in humans
and neonatal days occur during days 1-14 in rats and mice, whereas
guinea pigs behave more like humans).

Finally, screening for teratogenic events should also include exposure
of females before mating or shortly following mating.
Such a regimen is fruitful inasmuch as environmental
agents cause adverse effects.
Publication Types: Review Review, Tutorial PMID: 11572272

Discussion and Analysis of the Papers:

FA was distributed to all organs in the adult, the placenta and
fetus (Table 1), which was similar to that reported in male F344 rats,
guinea pigs and monkeys. (25,26).

The major difference is that the Japanese demonstrated the
incorporation of FA and its metabolites into the placenta and fetus.
The quantity of radioactivity remaining in maternal and fetal tissues
at 48 hours was 26.9% of the administered dose.

The DNA fraction contained 20 % and 50% of total incorporated
radioactivity in the maternal and fetal liver at 6 and 24 hours when
compared to the acid insoluble fraction (Fig. 1).

Of primary interest is that the incorporated radioactivity persisted
longer in the fetal liver and brain when compared to the mothers.

Also, since FA is a precursor of a number of biological compounds, it
would have been of prime interest to determine what fraction resulted
from either metabolic incorporation or from chemical reactivity of FA
(e.g. crosslinks, adduction, methylation) with biological molecules
(DNA, proteins, polypeptide, amino acids, etc.).

FA undergoes addition (adducts and alkylation) and condensation
(methene bridges) reactions with proteins and amino acids (27) as well
as nucleic acids and nucleosides/tides. (28)
It is a mutagen, crosslinking agent and an immunogen (28-30).
Free FA concentrations in the blood are 2.24+- 0.07 (rats), 1.84+- 0.15
(Rhesus monkeys) and 2.61+- 0.14 (humans) ug/g of blood [ppm],
which did not change following either acute or subchronic inhalation of
FA. (31,32)

Thus, it appears that additional information is required on addition
and condensation products of amino acids, polypeptides,
nucleoside, etc., of the blood, generated by FA exposure.

An increase of N-methyl amino acids would produce endogenous FA, which
may have a significant role in mitotic and apoptosis processes.

FA generators are responsible for FA formation in tumors and have an
impairment of liver antioxidant mechanisms and functional integrity of
mitochondria. (33-41)

FA had adverse effects on zygotes/embryos and bone marrow cells (Tables
2 and 3). The embryos showed cytological injury and high rate of
mortality, while bone marrow cells had increased rates of chromosome
aberrations and aneuploidy.

Similar observations on chromosomes of peripheral lymphocytes have been
reported for anatomy and mortuary students. (42-44)
Classroom exposure to FA at 1.5 to 3.17 mg/m3 was associated with
increased frequency of sister chromatid exchanges, aberrations and
micronuclei.
Concentrations less 1 mg/m3 had no effect on lymphocyte chromosomes,
but caused micronuclei in nasal and oral exfoliative cells
and changes in lymphocyte subsets (increase in CD19 and decreases in
CD4, CD5 and H/S ratio. (45,46)

With respect to the effect of FA on embryos additional research is
needed.

FA is an alkylating agent. Treatment of C3H transplacentally
with N-ethyl-N-nitrosourea (alkylating agent) has caused
primordial germ cell mutations. (47)

Also, treatment of female mice within hours after mating with ethyl
methanesulfanate, ethyl nitrosourea and ethylene oxide resulted
in fetal deaths and malformations. (48-51)

Thus, further investigation into the zygote/embryonic effects of FA
should follow the protocols established for other alkylating agents
with attention to the role of potential methyl donors,
e.g. N-methyl amino acids.

FA exposure throughout gestation caused a decreased DNA and RNA
concentrations, increased weights of bodies and organs (thymus, heart,
kidneys and adrenals) and decreased in the weights of lung and liver
(Table 3).

Microscopy and histochemical observations revealed other
abnormalities: involution of lymphoid tissue, numerous extra-medullary
hemopoietic centers, decreased glycogen content (myocardium) and liver,
decreased AA content of whole fetus and fetal and maternal liver.

AA is an antioxidant, produced from glucuronate via the uronic acid
pathway, which also is the intermediary route for synthesis of pentoses.

The decreased AA content may have resulted from either the utilization
of AA as an antioxidant or by interference (inhibition?) of the uronic
pathway.

It is difficult to interpret the meaning of the decreased DNA and the
increased RNA contents of the organs.
However, treatment of adult male rats by FA injection was reported to
decrease the DNA content of testis and prostate and a decrease of
protein content of the prostate and epididymus. (52)

Cytopathology of organs and alterations of mitochondria,
ER and lysosome enzymatic were observed in fetuses following FA
inhalation (Table 4).
Organ cytopathology included increased ploidy, micronecrotic loci,
extramedullary hematopoeitec enters, and degeneration of kidney
glomeruli. Concomitant were changes in enzymatic activity of as follows:

mitochondria (MDH, SDH, LDH decreased, while GDH increased);
ER and lysosomes (ATPase increased while inosine diphosphatase and
b-glucorinidase decreased).
The impairment lasted in the organs to 4 months of age.
In addition, N-acetylneuraminic concentration increased in maternal and
fetal tissues. The changes in the enzymatic activity and
N-acetyleneuraminic acid correlated with increased fetal mortality.
Finally, the development of postnatal behavior was also adversely
affected (Table 6).

FA has effects on mitochondrial enzymes, glutathione concentrations and
bile production in the liver of many species, including humans. (53)

FA inhibits the uptake of phosphate by mitochondria, (54,55) and causes
the release of GPT, SDH, GSSG and malondialdehyde into the perfusate of
isolated livers. (56)

Intraperitoneal injection results in a 2-fold increase in bile and a
significant decrease in glutathione of the liver, lungs and brains. (57)

An electron microscopic investigation of the perfused isolated livers
showed destruction of the mitochondria (ruptured membranes, loss of the
cristae) and some damage to the endoplasmic reticulum. (56)

The protection of the liver from FA toxicity appears to be dependent
upon glutathione by formation of the adduct S-hydroxymethylglutathione.
(58)

Thus, the observed effect of FA on mitochondrial and ER functions
during embryo/fetal development is also demonstrable in the adult liver.

FA caused preimplantation, prenatal and postnatal abnormalities.

The prenatal effects were demonstrable as anomalies and aberrancies in
blood buffering capacity with metabolic (formate?) acidosis.

The major anomalies were an increased frequency of cryptochordism, a
decrease/delay in ossification centers of the hyoid, metacarpus and
metatarsal bones, delay in eruption of incisors and a decrease in body
weight.

Blood pH decreased in the fetus, while the pCO2 (hypercapnia) increased
in the fetus and the mother.
The true bicarbonates and CO2 were unaffected by FA alone, but increased
with iron-deficiency in the fetus and mother.

The presence of iron-induced deficiency augmented these abnormalities,
along with increased embryo mortality .

The postnatal effect of FA was tested by maze performance.
Open field tests demonstrated an increase in motor activity, increase in
standing and appearance of emotion.
In sexually mature rats there was an increase in search activity.

FA is metabolized to formate.
Alcohols, particularly methanol and ethanol, are metabolized to formate
and lactate via an aldehyde.

The toxicity of alcohols and formalin in humans and animals includes
metabolic acidosis (59-61).

Alcohol toxicity generates free radicals,
cause an increase in malondialdehyde, and induce lipid peroxidation
resulting in DNA single strand breaks (62-66).

FA and alcohols probably affect embryos and the fetus via mitochondrial
damage.

Ethanol and environmental agents trigger apoptotic neurodegeneration in
the developing brain (67,68).

Oxygen stress, such as that caused by free radical generation, is
associated with apoptotic cell death and fragmentation of mitochondrial
genome (69-71).

Moreover, FA via formaldehyde generators, e.g. alkylating agents,
initiates apoptosis (72-74).
Mitochondria are the suicide organelles and control apoptosis (75-78).

Thus, subtle birth defects (autism, low birth weight, fetal alcohol
syndrome, etc.) are probably best understood by investigating in utero
oxidative stress and mitochondrial damage, rather than by standard FA
teratogenic research (79-83). ]



[ "Rats that had received aspartame (250 mg/kg/day)
[ 27.5 mg methanol,
and at 30%, 8.25 mg formaldehyde and formic acid retention ]
in the drinking water for 3 or 4 months showed a significant increase
in time to reach the reward in the T-maze,
suggesting a possible effect on memory due to the artificial sweetener."

This extraordinary result demands replication, because it implies a much
lower No Observed Effect Level (NOEL), which is customarily divided by
1000 to set an Acceptible Daily Limit for humans.

A formaldehyde NOEL of about 8 mg/kg bw/day would set a human
formaldehyde ADL much lower at .008 mg/kg bw/day,
which for a 60 kg person would be 0.5 mg daily,
if 30% retention of formaldehyde and formic acid durable cumulative
toxic products is allowed.

Recall that 12 cans daily diet soda gives 2400 mg aspartame,
264 mg methanol, and at 30% retention of formaldehyde and formic acid
cumulative durable toxic products, 80 mg daily, 160 times the ADL
suggested by the results of Christian et al, 2004.


http://groups.yahoo.com/group/aspartameNM/message/1088
chronic aspartame in rats affects memory, brain cholinergic receptors,
and brain chemistry, Christian B, McConnaughey M et al, 2004 May,
full plain text & critique: Murray 2004.06.05

reference 52:
Pharmacol Biochem Behav. 2004 May; 78(1): 121-7.
Chronic aspartame affects T-maze performance, brain cholinergic
receptors and Na(+),K(+)-ATPase in rats.
Christian B, McConnaughey K, Bethea E, Brantley S,
Coffey A, Hammond L, Harrell S, Metcalf K, Muehlenbein D,
Spruill W, Brinson L, McConnaughey M.
Department of Pharmacology, Brody School of Medicine,
East Carolina University, Greenville, NC 27858, USA;
North Carolina School of Science and Mathematics,
Durham, NC 27811.
http://www.ecu.edu/pharmacology/faculty/mcconnaughey.html
Mona M. McConnaughey, Ph.D. Research Assistant Professor
Department: PHARMACOLOGY & TOXICOLOGY
Office: Brody Medical Science 6E-120A 252-744-2756
MCCONNAUGHEYM@...

This study demonstrated that chronic aspartame consumption in rats
can lead to altered T-maze performance and increased muscarinic
cholinergic receptor densities in certain brain regions.
Control and treated rats were trained in a T-maze to a particular side
and then periodically tested to see how well
they retained the learned response.

Rats that had received aspartame (250 mg/kg/day)
[ 27.5 mg methanol, and at 30%, 8.25 mg formaldehyde and formic acid
retention ]
in the drinking water for 3 or 4 months showed a significant increase
in time to reach the reward in the T-maze,
suggesting a possible effect on memory due to the artificial sweetener.

Using [(3)H]quinuclidinyl benzilate (QNB) (1 nM) to label muscarinic
cholinergic receptors and atropine (10(-6) M) to determine nonspecific
binding in whole-brain preparations,
aspartame-treated rats showed a 31 % increase in receptor numbers
when compared to controls.
In aspartame-treated rats, there was a significant increase in
muscarinic receptor densities in the
frontal cortex, midcortex, posterior cortex, hippocampus, hypothalamus
and cerebellum of
80 %, 60 %, 61 %, 65 %, 66 % and 60 %, respectively.
The midbrain was the only area where preparations from
aspartame-treated rats showed a significant increase
in Na(+),K(+)-ATPase activity.
It can be concluded from these data that long-term consumption
of aspartame can affect T-maze performance in rats and alter
receptor densities or enzymes in brain. PMID: 15159141

[ The ASE review here concludes with a grudging admission:

"There are two studies that,
when using only one dose and only one measure of learning,
interpreted their findings to indicate an impairment of learning by
aspartame at doses of 250 and 500 mg/kg/day." ]

" 6.4.2.3 Recently, Christian et al. (2004) used a T-maze to test for
memory loss in male Sprague-Dawley rats receiving aspartame (250 mg/kg
bw/day) in drinking water for 120 days.

Twelve rats per group were tested periodically during the treatment
period for the time required to find a reward (chocolate) in the T-maze.

Rats were tested every 2 weeks. Although no differences were noted
during the measurements occurring approximately every 2 weeks up to
about 72 weeks, rats receiving aspartame took significantly longer to
find the reward after 90 days.

The next measurement shown is at 120 days, when rats again took longer
to find the reward.

Rats were then killed and levels of brain cholinergic receptors and
NaK-ATPase levels were determined.

Muscarinic cholinergic receptor density, measured using radiolabeled
quinuclindinyl benzilate, was significantly higher in several areas of
the brain of rats receiving aspartame compared to rats receiving only water.

NaK-ATPase levels were similar in all areas of the brain, except for the
midbrain where levels were higher in aspartame-treated rats.

The authors (Christian et al., 2004) speculate that the increase in time
to find a reward in the T-maze indicates that chronic aspartame
consumption results in memory loss and this was mediated by the change
in cholinergic receptor density.

However, they also acknowledge that the relationship between memory and
receptor density is not well understood.

The authors suggest that an alternative explanation for the increased
time in the T-maze test is that rats treated with aspartame may have a
reduced desire for the chocolate reward.

As rats do not recognize aspartame as a sweet substance, it is not clear
how this may occur.

One possibility is that as aspartame is also used to modify the flavors
of other substances (when given below the sweetness threshold in
humans), it is possible that while the rats might not taste aspartame as
sweet, aspartame could have modified the taste of the chocolate (e.g.,
making it more bitter).

Major limitations of this study are the use of only one tool or task to
measure memory and the use of only one dose of aspartame.

The majority of studies designed to detect an effect of aspartame on
learning or memory have used multiple doses and multiple tests to
evaluate these parameters.

These studies report no effect, at doses as high as 4% aspartame in the
diet, even when exposed from conception to 90 days postnatally.

Higher doses have been reported to affect various indices of learning
behavior.

There are two studies that,
when using only one dose and only one measure of learning,
interpreted their findings to indicate an impairment of learning by
aspartame at doses of 250 and 500 mg/kg/day.

In summary, well-designed studies using a range of approaches to
evaluate learning and memory consistently demonstrate no effect of
aspartame consumption at levels up to 4000 mg/kg/day, indicating little
likelihood that aspartame will have an effect on memory or learning in
humans. "


6.4.2.4. Effects on Aggression.

Male Long-Evans rats received single intraperitoneal injections of 0,
200, 400, and 800 mg/kg bw aspartame in vehicle (saline plus Tween 80)
and then were exposed to an intruder rat and observed for aggression
(Goerss et al., 2000).

Eleven rats were tested with all aspartame doses delivered in ascending
then descending order with a minimum of 1 day between tests.

Contrary to an expected enhancement of aggressive behavior based on the
hypothesis that aspartame is neuroexcitatory, when rats received
aspartame doses of 200 mg/kg bw or higher,
the time to attack an intruder rat increased,
the number of bites decreased,
and fewer rats engaged in aggressive attacks.

Rats treated with 200 and 400 mg/kg bw aspartame had higher levels of
serotonin (5-hydroxytryptamine) and its metabolite, 5-hydroxyindolacetic
acid, in the brain compared to rats receiving vehicle, but this effect
was not evident in rats treated with 800 mg/kg bw aspartame.

Dopamine and its metabolites, dihydroxyphenylacetic acid and
homovanillic acid, were not affected by aspartame treatments.

The authors (Goerss et al., 2000) were unable to explain how aspartame
administration would result in increased serotonin levels in these groups.

The rationale for intraperitoneal administration of aspartame was not given.

An intraperitoneal exposure is an irrelevant route of administration
when aspartame is extensively metabolized prior to absorption, and the
only human exposure is via oral administration.

In summary, there has been extensive investigation of the possibility of
neurotoxic effects due to consumption of aspartame.

The data from these studies, in general, do not support the hypothesis
that aspartame in the human diet will affect neuronal function, learning
or behavior.

[ This 2000 study demands careful replication, using even lower oral
doses, determination of the actual disposition of toxic metabolites
in specific tissues, and more tests of behavioral changes. Dimethyl
dicarbonate would make an ideal control.

It's possible that the toxic effects of intraperitoneal injection of
aspartame were depressing the rat's overall evergy level.

Anxiety, restlessness, and irritability, or depression, fatigue, and
dulled affect are major symptoms for aspartame reactors and alcohol
hangover.

Behavioral studies may confirm the very low ADL of 4.8 mg formaldehyde
for a 60 kg person, as reasonably suggested by the results from
Christian et al, 2004.

It is not surprising that researchers are not readily funded to do
simple, low cost, fast studies that would compel steep limitations on
billions of dollars annual business of world vested interests for
products ranging from diet and alcohol beverages to buildings and mobile
homes to methanol vehicle fuels, and perhaps an entire new field of
concerns about the otherwise ideal fruits and vegetables. ]


6.3.2 Lifetime Studies

[ The ASE review has devoted section 6.3.2 to many detailed criticisms
of the two Ramazzini lifetime cancer rat studies.

As a medical layman, never educated in biochemistry, I am not qualified
to address these matters, and I welcome comments pro and con, which I
will put on my aspartameNM group public archive:

aspartameNM@yahoogroups.com

I am qualified, however, to remind scientists that effective
collaboration between opposed viewpoints has to based on an ethic of
going out of one's way to present the best expressions by others of
their evidence.

In fact, the Ramazzini Foundation has a large network of eminent
supporters, some of whom have taken an unusual public position in
support of their two cancer studies:

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


In addition, Ramazzini Foundation proved carcinogenity by similar
studies in Dec., 2002 for alcohol, aldehyde, methanol, and formaldehyde,
and the results are very compatible with their first aspartame cancer
study, 2005, and with their second aspartame study, 2007:

http://groups.yahoo.com/group/aspartameNM/message/1186
aspartame induces lymphomas and leukaemias in rats,
free full plain text, M Soffritti, F Belpoggi, DD Esposti, L Lambertini,
2005 April, 2005.07.14: main results agree with their previous methanol
and formaldehyde studies, Murray 2005.07.19

""Yellowing of the coat was observed in animals exposed to APM, mainly
at the highest concentrations.

This change was previously observed in our laboratory in rats exposed
to formaldehyde administered with drinking water 9."

http://groups.yahoo.com/group/aspartameNM/message/1441
Lifetime exposure to low doses of aspartame beginning during prenatal
life increases cancer effects in rats, Morando Soffritti et al, European
Ramazzini Foundation, USA EPA Environmental Health Perspectives
2007.06.13 free full text 24 pages: Murray 2007.06.16
www.ehponline.org/members/2007/10271/10271.pdf free full text 24 pages

9. Soffritti M, Belpoggi F, Lambertini L, et al.
Results of longterm experimental studies on the carcinogenicity of
formaldehyde and acetaldehyde in rats. In
Mehlman MA, Bingham E, Landrigan PJ, et al.
Carcinogenesis bioassays and protecting public health.
Commemorating the lifework of Cesare Maltoni and colleagues.
Ann NY Acad Sci 2002; 982: 87-105.

11. Soffritti M, Belpoggi F, Cevolani D, et al.
Results of long-term experimental studies on the carcinogenicity of
methyl alcohol and ethyl alcohol in rats. In
Mehlman MA, Bingham E, Landrigan PJ, et al.
Carcinogenesis bioassays and protecting public health.
Commemorating the lifework of Cesare Maltoni and colleagues.
Ann NY Acad Sci 2002; 982: 46-69.


Ann N Y Acad Sci. 2002 Dec; 982: 87-105.
Results of long-term experimental studies on the carcinogenicity of
formaldehyde and acetaldehyde in rats.
Soffritti M, Belpoggi F, Lambertin L, Lauriola M,
Padovani M, Maltoni C. crcfr@...
Cancer Research Center, European Ramazzini Foundation
for Oncology and Environmental Sciences, Bologna, Italy.

Formaldehyde was administered for 104 weeks in drinking water
supplied ad libitum at concentrations of
1500, 1000, 500, 100, 50, 10, or 0 mg/L
to groups of 50 male and 50 female Sprague-Dawley rats
beginning at seven weeks of age.
Control animals (100 males and 100 females)
received tap water only.
Acetaldehyde was administered to 50 male and 50 female
Sprague-Dawley rats beginning at six weeks of age
at concentrations of
2,500, 1,500, 500, 250, 50, or 0 mg/L.
Animals were kept under observation until spontaneous death.
Formaldehyde and acetaldehyde were found to produce
an increase in total malignant tumors in the treated groups
and showed specific carcinogenic effects
on various organs and tissues. PMID: 12562630

Ann N Y Acad Sci. 2002 Dec; 982: 46-69.
Results of long-term experimental studies on the carcinogenicity of
methyl alcohol and ethyl alcohol in rats.
Soffritti M, Belpoggi F, Cevolani D,
Guarino M, Padovani M, Maltoni C.
Cancer Research Center,
European Ramazzini Foundation for Oncology and
Environmental Sciences, Bologna, Italy. crcfr@...

Methyl alcohol was administered in drinking water supplied ad libitum
at doses of
20,000, 5,000, 500, or 0 ppm to groups of male and female
Sprague-Dawley rats 8 weeks old at the start of the experiment.
Animals were kept under observation until spontaneous death.
Ethyl alcohol was administered by ingestion in drinking water at a
concentration of 10% or 0% supplied ad libitum to groups of male and
female Sprague-Dawley rats; breeders and offspring were included in the
experiment.
Treatment started at 39 weeks of age (breeders), 7 days before mating,
or from embryo life (offspring) and lasted until their spontaneous death.
Under tested experimental conditions, methyl alcohol and ethyl alcohol
were demonstrated to be carcinogenic for various organs and tissues.
They must also be considered multipotential carcinogenic agents.
In addition to causing other tumors, ethyl alcohol induced malignant
tumors of the oral cavity, tongue, and lips.
These sites have been shown to be target organs in man by epidemiologic
studies. Publication Types: Review Review, Tutorial PMID: 12562628


Here I have combined fairly equivalent data from their aspartame,
methanol, and formaldehyde studies. Aspartame groups were 100-150 rats
each, methanol 100 rats each, and formaldehyde 50 rats each
(formaldehyde control groups 100 rats each).

Aspartame and methanol are directly comparable, since the 11% methanol
component of aspartame upon ingestion is immediately and fully released
into the GI tract, and then much of that quickly turned into
formaldehyde and then formic acid, both of which account for the
toxicity of methanol.


Comparison of two aspartame, one methanol, one formaldehyde studies:

Males
Females
Males + Females

Animals with lymphomas and leukaemias [hemolymphoreticular
neoplasias] % of each group of animals

Group
100 rats each
70 rats each 2nd cancer study 2007

aspartame dose ppm a
[400 ppm in 20 gm feed = 20 mg/kg rat body weight for 0.4 kg rats)

--------equivalent methanol dose (11% of aspartame)
----------------roughly equivalent formaldehyde dose (30% of methanol)

--------------------20,000-40.0
---------------------------28.0 #^
-------------------------- 34.0

I--100,000-29.0
-----------25.0**
-----------27.0

II--50,000-0.0-------5,000-36.0---1,500-46.0 **
-----------25.0**----------24.0---------20.0*
-----------22.5------------30.0---------33.0

----------------------------------1,000-22.0*
----------------------------------------22.0*
----------------------------------------22.0

------------------------------------500-24.0*
----------------------------------------14.0
----------------------------------------19.0

III-10,000-15.0
-----------19.0*
-----------17.0

-----------------------500-35.0
---------------------------24.0
---------------------------29.5

-----------------------100-26.0**
---------------------------16.0
---------------------------21.0

-------------------------------------50-20.0
----------------------------------------14.0
----------------------------------------17.0

IV----2,000-22.0
------------18.7*
------------20.3

------2,000-17.1 70 rats, 2nd study 2007
------------31.4
------------24.3


V-------400-16.7
------------20.0**
------------18.3

--------400-15.7 70 rats, 2nd study 2007
------------17.1
------------16.4


-------------------------------------10--8.0
----------------------------------------10.0
-----------------------------------------9.0

-----------------------15-20.0 [-50 rats ]
--------------------------10.0 [-50 rats ]
--------------------------15.0 [100 rats ]

VI--------80-15.3
-------------14.7
-------------15.0

VII--------0-20.7--------0-28.0--------0--8.0 [ control groups ]
--------------8.7----------13.0-----------7.0
-------------14.7----------20.5-----------7.5

-----------0--9.5 2nd cancer study 2007 95 rats each control group
-------------12.6
-------------11.0 190 rats, combined male and female control groups

a ppm Considering the life-span average weight of a rat (male and
female) as 400 g and the average consumption of food as 20 g per day

* aspartame, statistically significant p= 0.05;
** aspartame, statistically significant p= 0.01
using poly-k test (k = 3)

# methanol, p<0.05 using X2 test
^ methanol, p<0.05 using Cochrane-Armitage test for dose-response
relationship

* formaldemyde, p<0.05 using X2 test
** formaldehyde, p<0.01 using X2 test

The control groups vary widely, with the percentage of rats with these
most common cancers, present at natural death,
ranging from 7.0% to 28.0%.

A layman can only speculate as to the possible causes in a uniform
population of rats in the same huge laboratory facility for decades,
such as various viruses, bacteria, or molds, or variable impurities in
the tap water.

Formaldehyde at 50 ppm shows a doubling of the percentage of rats with
these cancers, for groups of just 50 rats.

It is a safe bet that studies using groups of 100 to 200 rats would
establish significance at this 50 ppm level, which in turn would mandate
the reduction of the present USA EPA level (1999) from 1 ppm
for lifetime exposure to formaldehyde in drinking water to 0.05 ppm,
since the human limit is estimated by dividing the lowest harmful animal
level by 1000.

The various standards for methanol and formaldehyde are not in harmony:

We can grasp the main picture by studying the results at a high level of
exposure:

II--50,000-20.0------5,000-36.0-1,500--46.0**
-----------25.0**----------24.0--------20.0*
-----------22.5------------30.0--------33.0

The results amount to 1.3 to 5.75 times their control group levels.
Aspartame, methanol, and formaldehyde results broadly agree.
Unknown factors are causing differences between males and females.



6.6.1 "Mice Male ddY mice 2000 mg/kg bw Negative in Comet
assay Sasaki et al. (2002)"

6.6.2 "Sasaki et al. (2002) assessed the genotoxicity of 39 food
additives using the comet assay of tissues collected from male ddY mice
administered 1 oral dose at up to one half of the LD50 or 2000 mg/kg bw.

For aspartame, a dose of 2000 mg/kg bw was used and mice (n = 4/group)
were killed after 3 and 24 h.

Nuclei were obtained from glandular stomach, colon, liver, kidney,
urinary bladder, lung, brain, and bone marrow.

No changes were found in the comet lengths of nuclear DNA from tissues
from aspartame-treated mice.

Thus, aspartame was found to be nongenotoxic in this assay."

[ The ASE review missed the actual details of this provokative study by
an experienced team that continues to use the sensitive, fast, low cost
automated Comet assay to measure DNA damage.

The team tested 39 food additives, including 7 sweeteners, using
groups of just 4 mice at a uniform oral dose of 2000 mg/kg bw, testing 8
organs at 3 or 24 hours later.

However, each test group had an assigned control group, somehow selected
from a total of 21 4-rat control groups.

Although the aspartame control levels happened to be much higher than
the averages for all 21 control groups, the aspartame values were about
twice the control values, close to statistical significance, 3 hours
after the oral dose, for stomach, colon, liver, bladder, and lung.

Results for DNA damage for Ace-K and stevia were nil, while sucralose,
cyclamate, and saccharin were significant for many tissues.

This result, once again, cries out for thorough replication, with much
larger groups of mice, using a variety of dose levels and test times,
and testing many more tissues.

A separate report confirmed that stevia was not genotoxic. ]

http://groups.yahoo.com/group/aspartameNM/message/935
Comet assay finds DNA damage from sucralose, cyclamate, saccharin
in mice: Sasaki YF & Tsuda S Aug 2002: Murray 2003.01.01
[ Also borderline evidence, in this pilot study of 39 food additives,
using test groups of 4 mice, for DNA damage from for stomach, colon,
liver, bladder, and lung 3 hr after oral dose of 2000 mg/kg aspartame --
a very high dose. Methanol is the only component of aspartame
that can lead to DNA damage. ]


http://groups.yahoo.com/group/aspartameNM/message/961
genotoxins, Comet assay in mice: Ace-K, stevia fine; aspartame poor;
sucralose, cyclamate, saccharin bad: Y.F. Sasaki Aug 2002:
Murray 2003.01.27 [A detailed look at the data] ]

"Comparing the mean control values [average of all 21 control groups] to
the values for the other 7 sweeteners:

Best is acesulfame K, with no significant or high values.

Good is glycyrrhizin (derived from licorice), two 1.4 ratios for Stomach
and Brain.

Next is stevia, with one high value [above ratio 1.4],
9.48+-1.99 for Bladder, 2000 mg 3 hr, ratio 1.8 .

Aspartame has high values for 2000 mg 3 hr for Stomach, Colon, Liver,
Bladder, Lung.

Sucralose has 3 significant values and 13 high values, for Stomach,
Colon, Kidney, Bladder, Lung, Brain.

Sodium cyclamate has 4 significant values and 10 high values for
Stomach, Colon, Liver, Kidney, Bladder, Lung, Brain, Bone.

Saccharin has 3 highly significant values for Colon, and 13 high values
for Stomach, Colon, Kidney, Lung, Brain, Bone.

Sodium saccharin has 5 highly significant values for Stomach and Colon,
and 14 high values for Stomach, Liver, Kidney, Bladder, Lung, Brain,
Bone."


"For Liver, 5 of the 21 control groups, with values 1.67, 1.63, 1.29,
1.06, 1.65 would make some 3 hr aspartame values approach or reach
significance.

Ratios about 2 for different tissues with aspartame that would be close
to significant would exist for many of the 21 control groups:
Stomach 1 Colon 5 Liver 5 Bladder 11 Lung 5 .

The aspartame values at 3 hr are compared with
the mean values for the 21 control groups:

Somach ---- Colon ---- Liver ---- Kidney ---- Bladder ---- Lung

DNA Migration at 3 hr from 2000 mg/kg dose
8.49+-0.48, 9.18+-0.56, 3.26+-0.16, 1.91+-0.26, 10.7+-2.77, 4.13+-1.26

mean of 21 control groups
6.31,------ 5.81,------ 2.15,------ 2.25, ------ 5.40, ----- 2.61,

range of values for 21 control groups
4.3-8.6 --- 4.0-8.1 --- 1.1-3.6 --- 1.2-2.9 ---- 3.6-7.1 --- 1.6-4.7

ratio = DNA Migration/control mean
1.4, ------ 1.6, ------ 1.5, ------ 0.9, ------- 2.0, ------ 1.6

Brain ---------- Bone [marrow]

0.37+-0.70, ---- 1.01+-0.59 DNA Migration at 3 hr from 2000 mg/kg dose


1.48, ---------- 1.12, mean of 21 control groups


0.8-2.6 -------- 0.6-1.9 range of values for 21 control groups


0.3, ----------- 0.9, ratio = DNA Migration/control mean

Wouldn't the average of all the 21 control groups be the best control
values to use?

What would then be the appropriate statistical test?

How many mice would it take to reach significance for the 5 tissues with
ratios over 1.4: Stomach, Colon, Liver, Bladder, Lung?

Aspartame at 24 hours had levels too low to reach significance with any
of the 21 control groups."


reference 306:
Mutat Res 2002 Aug 26; 519(1-2): 103-19
The comet assay with 8 mouse organs:
results with 39 currently used food additives.
Sasaki YF, Kawaguchi S, Kamaya A, Ohshita M,
Kabasawa K, Iwama K, Taniguchi K, Tsuda S.
Laboratory of Genotoxicity, Faculty of Chemical and Biological
Engineering, Hachinohe National College of Technology,
Tamonoki Uwanotai 16-1, Aomori 039-1192, Japan.
yfsasaki-c@... ; s.tsuda@...

We determined the genotoxicity of 39 chemicals
currently in use as food additives.
They fell into six categories-dyes, color fixatives and
preservatives, preservatives, antioxidants, fungicides, and sweeteners.

We tested groups of four male ddY mice once orally with each additive
at up to 0.5xLD(50) or the limit dose (2000 mg/kg) and performed the
comet assay on the glandular stomach, colon, liver, kidney,
urinary bladder, lung, brain, and bone marrow
3 and 24 h after treatment.

Of all the additives, dyes were the most genotoxic.
Amaranth, Allura Red, New Coccine, Tartrazine, Erythrosine,
Phloxine, and Rose Bengal i
nduced dose-related DNA damage in the glandular stomach, colon,
and/or urinary bladder.
All seven dyes induced DNA damage in the gastrointestinal organs
at a low dose (10 or 100 mg/kg).

Among them, Amaranth, Allura Red, New Coccine, and Tartrazine
induced DNA damage in the colon at close
to the acceptable daily intakes (ADIs).

Two antioxidants (butylated hydroxyanisole (BHA)
and butylated hydroxytoluene (BHT)), three fungicides
(biphenyl, sodium o-phenylphenol, and thiabendazole),
and four sweeteners (sodium cyclamate, saccharin, sodium saccharin,
and sucralose) also induced DNA damage in gastrointestinal organs.

Based on these results, we believe that more extensive assessment of
food additives in current use is warranted. PMID: 12160896


A single can of diet soda gives 200 mg aspartame,
22 mg methanol, and toxic products 6.6 mg, 13 times above the possible ADL.

Accordingly, we should give much more respectful attention to the
thousands of case reports of toxic symptoms, especially neurological, by
professionals and citizens:

http://www.dorway.com/tldaddic.html 5-page review full text
Roberts HJ Aspartame (NutraSweet) addiction.
Townsend Letter 2000 Jan; HJRobertsMD@...
http://www.sunsentpress.com/ sunsentpress@...
Sunshine Sentinel Press P.O.Box 17799 West Palm Beach, FL 33416
800-814-9800 561-588-7628 561-547-8008 fax

http://groups.yahoo.com/group/aspartameNM/message/669
1038-page medical text "Aspartame Disease: An Ignored Epidemic"
published May 30 2001 $ 60.00 postpaid data from 1200 cases
available at http://www.amazon.com
over 600 references from standard medical research

http://groups.yahoo.com/group/aspartameNM/message/790
Moseley: review Roberts "Aspartame Disease: An Ignored Epidemic":
Murray 2002.07.02

http://groups.yahoo.com/group/aspartameNM/message/883
three texts by H.J. Roberts, 1958, 1971, 1979: Murray 2002.11.06

http://groups.yahoo.com/group/aspartameNM/message/880
Roberts 45 clinical research reports in mainstream journals:
Murray 2002.10.20 ]



[ The following critiques will be appreciated by the few who have the
tenacity to plow through the tedious mazes of red herrings, omitted
studies, incuriosity, bias, cursory dismissals, unexamined details,
ignored opportunities, spiced with sonorous PR spin:
"...the most tested food additive in history,"
"...used in over 6,000 products worldwide,"
"...approved by FDA, WHO, EU FSA, American Diabetes Assc., ect,"
"...components are part of normal foods and metabolism in much greater
amounts,"
"...finally put to rest all the rumors on the Net, using a tiny minority
of allergic individuals to opportunistically impugn distingished
experts, who are valiantly protecting diabetics and the obese against
the ravages of sugar."

http://groups.yahoo.com/group/aspartameNM/message/1108
faults in 1999 July EPA 468-page formaldehyde profile:
Elzbieta Skrzydlewska PhD, Assc. Prof., Medical U. of Bialystok, Poland,
abstracts -- ethanol, methanol, formaldehyde, formic acid, acetaldehyde,
lipid peroxidation, green tea, aging: Murray 2004.08.08 2005.07.11

http://groups.yahoo.com/group/aspartameNM/message/1070
critique of aspartame review, French Food Safety Agency AFSSA
2002.05.07 aspartamgb.pdf (18 pages, in English), Martin Hirsch:
Murray 2004.04.13

http://groups.yahoo.com/group/aspartameNM/message/957
safety of aspartame Part 1/2 12.4.2: EC HCPD-G SCF:
Murray 2003.01.12 EU Scientific Committee on Food, a whitewash

http://groups.yahoo.com/group/aspartameNM/message/1045
http://www.holisticmed.com/aspartame/scf2002-response.htm
Mark Gold exhaustively critiques European Commission Scientific
Committee on Food re aspartame ( 2002.12.04 ):
59 pages, 230 references

http://www.eatright.org/Nutritive(1).pdf
J Am Diet Assoc. 2004 Feb; 104(2): 255-75.
Position of the American Dietetic Association: use of nutritive and
nonnutritive sweeteners. American Dietetic Association.

http://groups.yahoo.com/group/aspartameNM/message/1068
critique of aspartame review
by American Dietetic Association Feb 2004,
Valerie B. Duffy & Madeleine J. Sigman-Grant: Murray 2004.05.14


[ Here's a useful list... ]

reference 42:
Regul Toxicol Pharmacol. 2002 Apr; 35(2 Pt 2): S1-93.
Aspartame: review of safety.
Butchko HH,
Stargel WW,
Comer CP,
Mayhew DA,
Benninger C,
Blackburn GL,
de Sonneville LM,
Geha RS,
Hertelendy Z,
Koestner A,
Leon AS,
Liepa GU,
McMartin KE,
Mendenhall CL,
Munro IC,
Novotny EJ,
Renwick AG,
Schiffman SS,
Schomer DL,
Shaywitz BA,
Spiers PA,
Tephly TR,
Thomas JA,
Trefz FK.
Medical and Scientific Affairs, The NutraSweet Company, Mt Prospect,
Illinois 60056, USA. harriett.h.butchko@...

Over 20 years have elapsed since aspartame was approved by regulatory
agencies as a sweetener and flavor enhancer.

The safety of aspartame and its metabolic constituents was established
through extensive toxicology studies in laboratory animals, using much
greater doses than people could possibly consume.

Its safety was further confirmed through studies in several human
subpopulations, including healthy infants, children, adolescents, and
adults; obese individuals; diabetics; lactating women; and individuals
heterozygous (PKUH) for the genetic disease phenylketonuria (PKU) who
have a decreased ability to metabolize the essential amino acid,
phenylalanine.

Several scientific issues continued to be raised after approval, largely
as a concern for theoretical toxicity from its metabolic components --
the amino acids, aspartate and phenylalanine, and methanol -- even
though dietary exposure to these components is much greater than from
aspartame.

Nonetheless, additional research, including evaluations of possible
associations between aspartame and headaches, seizures, behavior,
cognition, and mood as well as allergic-type reactions and use by
potentially sensitive subpopulations, has continued after approval.

These findings are reviewed here.

The safety testing of aspartame has gone well beyond that required to
evaluate the safety of a food additive.

When all the research on aspartame, including evaluations in both the
premarketing and postmarketing periods, is examined as a whole, it is
clear that aspartame is safe, and there are no unresolved questions
regarding its safety under conditions of intended use.
PMID: 12180494


6.3.1 "6.3.1. Two-Year Bioassay Studies

Aspartame was first approved by FDA as a nonnutritive sweetener in 1974,
based on the toxicity studies that were conducted by the Searle
Laboratories.

Chronic toxicity studies with aspartame, and its decomposition product,
DKP, were conducted in mice, rats, and dogs.

A 46-week study with aspartame was also performed with hamsters.

Table 15 provides a summary of these studies.

The carcinogenic potential of these compounds is discussed in the next
sections.

Following the approval of aspartame, a formal objection was submitted to
the FDA (FDA, 1981) questioning the conclusions from the rodent studies
on aspartame conducted by Searle, and proposing that aspartame may have
the potential to cause brain tumors in humans.

This objection resulted in FDA staying the regulation approving the
marketing of aspartame in 1975, and the establishment of a Public Board
of Inquiry to reexamine the studies submitted by Searle to the FDA.

Prior to the evaluation by the Board, the 15 studies submitted by Searle
were thoroughly audited by the Universities Associated for Research and
Education in Pathology, Inc. (UAREP) and by the FDA.

The findings of the UAREP, the FDA, and the Public Board of Inquiry were
considered and evaluated by the Commissioner of Food and Drugs,
resulting in the issuance of the commissioner's Final Decision that at
projected levels of consumption, aspartame would not pose a risk of
brain damage and will not cause brain tumors (FDA docket, 75F-0355,
1981) (FDA, 1981).

This decision resulted in FDA vacating the stay of the original 1974
regulation.

Objections to the of the use of aspartame were again filed with the FDA
in 1983; however, the regulations approving the use of aspartame was not
stayed following these objections, as the FDA stated that they failed to
create sufficient doubt about the safety of aspartame.

A response to the objections and a denial for a hearing was issued in
1984 by the Acting Commissioner of Food and Drugs (FDA docket 75F-0355
and 82F-0305) (FDA, 1984; Wurtman and Maher, 1987). "

[ In fact the FDA brought suit against Searle for its radically biased,
improper scientific studies.

The industry won by persuading the FDA's two attorneys to let the legal
process languish.

Soon,the attorney's inexplicably found peasant, prosperous futures in
the industry's networks.

Similar adroit exercises in corporate realpolitic were led by the CEO of
Searle, none other than that modest American hero, Donald Rumsfeld,
who used Reagan's victory to immediately manipulate the FDA's approval
by a brand new Commisioner, Arthur Hull Hayes, in July, 1981, of
aspartame in dry foods, and soon in beverages two years later, whereupon
the fortunate Commissioner, troubled by hints of polical corruption,
found a gracious life with the industry's PR agency.

http://www.dorway.com/enclosur.html
http://groups.yahoo.com/group/aspartameNM/message/53
aspartame history Part 1/4 1964-1976: Gold: Murray 1999.11.06

http://www.dorway.com/upipart1.txt
http://groups.yahoo.com/group/aspartameNM/message/262
aspartame expose 96K Oct 1987 Part 1/3: Gregory Gordon,
UPI reporter: Murray 2000.07.10

http://groups.yahoo.com/group/aspartameNM/message/928
revolving door, Monsanto, FDA, EPA: NGIN: Murray 2002.12.23
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About Us
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www.reuters.com/article/healthNews/idUSN1141652720070912

Industry-funded study calls aspartame safe
Tue Sep 11, 2007 10:58PM EDT
By Will Dunham

WASHINGTON (Reuters) - A review of 500 studies conducted over a quarter
century has turned up no credible evidence that the widely used
artificial sweetener aspartame is unsafe, industry-funded research
released on Tuesday showed.

A panel of American, British and Dutch experts rejected the notion that
aspartame causes cancer, seizures, neurological damage or learning
problems, or contributes to obesity. The panel did conclude that some
people might be prone to headaches after consuming it.

The group did not conduct original research but assessed existing
studies on the safety of aspartame, first approved by the U.S. Food and
Drug Administration in 1981. It is widely used in diet soft drinks and
sold in packets for use in coffee, tea or on food.

The panel saw consumption of it increasing, but called it safe for
adults or children and even among the heaviest users.

"Controlled and thorough scientific studies confirm aspartame's safety
and find no credible link between consumption of aspartame at levels
found in the human diet and conditions related to the nervous system and
behavior, nor any other symptom or illness," the researchers said in a
paper published in the journal Critical Reviews in Toxicology.

"There is no credible evidence that aspartame is carcinogenic," they added.

Michael F. Jacobson, PhD, executive director of the consumer group
Center for Science in the Public Interest, said the study grossly
exaggerates the importance of studies doubting health risks while
rejecting studies identifying possible problems.

Jacobson said "clouds of doubt" linger about aspartame's safety and said
it would be wise to minimize consumption of it until the research is
definitive. "This review is totally unreliable and people should simply
ignore it," Jacobson said.

Since its approval, some people have argued that aspartame can cause a
variety of illnesses, and various Web sites such as www.sweetpoison.com
denounce the sweetener.

ITALIAN FINDINGS

The researchers rejected the findings of a study published in June by
Italian scientists that showed aspartame might cause leukemia, lymphoma
and breast cancer in rats. University of Maryland food and nutrition
professor Bernadene Magnuson said that study was undermined by "numerous
methodological and interpretation errors."

The FDA said after that study was published that it saw no reason to
alter its view that aspartame is safe.

Magnuson said one possible area of concern was research indicating some
may get headaches after consuming aspartame. Magnuson cited conflicting
results in headache studies, but said that "a small subset of the
population" may have some sensitivity to aspartame-induced headaches.

The panel reviewed more than 500 human and animal studies, articles and
reports performed in the past 25 years.

"Certainly it is one of the most studied compounds ever," panel chairman
Dr. William Waddell of the University of Louisville told reporters,
saying the panel's 98-page report should put to rest safety concerns
relating to aspartame.

The panel's work was funded by Japanese food and seasonings giant
Ajinomoto Co, a maker of aspartame. Magnuson said the company had no
control over who was on the panel or how their work was done, and panel
members did not know who was funding the work until it was done.

"I have no qualms in terms of who funded the study," Magnuson told
reporters.

Aspartame is used in more than 6,000 food products worldwide. Merisant
Co is another leading aspartame company, with the brands Equal, Canderel
and NutraSweet.
////////////////////////////////////////////////////////////


www.informaworld.com/smpp/section?content=a781888262&fulltext=713240928
$ 32

Aspartame: A Safety Evaluation Based on Current Use Levels, Regulations,
and Toxicological and Epidemiological Studies
1. INTRODUCTION
Preamble
2. DESCRIPTION, SPECIFICATIONS, AND MANUFACTURING PROCESS
3. ESTIMATED DAILY INTAKE
4. ABSORPTION, DISTRIBUTION, METABOLISM, AND ELIMINATION
5. BIOCHEMICAL EFFECTS
6. SAFETY EVALUATION
7. EVALUATION SUMMARY
8. CONCLUSIONS
ADDENDUM
APPENDIX I: ESTIMATED MAXIMUM DAILY INTAKE OF ASPARTAME FROM REQUESTED
FOODS *
APPENDIX II: METHANOL CONTENT OF FOODS AND BEVERAGES
APPENDIX III: FORMALDEHYDE CONTENT OF COMMON FOODS
APPENDIX IV: TRANSGENIC MICE MODELS USED BY THE NATIONAL TOXICOLOGY
PROGRAM FOR THE EVALUATION OF THE CARCINOGENICITY OF ASPARTAME
APPENDIX V: USE OF TRANSGENIC MODELS IN REGULATORY EVALUATIONS
ACKNOWLEDGMENTS
REFERENCES
Notes
List of Figures
List of Tables

Aspartame: A Safety Evaluation Based on Current Use Levels, Regulations,
and Toxicological and Epidemiological Studies

Authors:


Bernadene A. Magnuson a; [ Bernadene A. Magnuson
Assistant Professor, Nutrition and Food Science

Expertise Key Words:
Prevention of colon cancer by food components, including vitamins,
nutrients and non-nutrients, such as plant and spice compounds;
Safety assessment of foods, food ingredients and dietary supplements
using toxicology data and risk assessment principles.

Expertise Credentials:
Over 25 peer-reviewed publications and several patents in cancer
prevention research and food toxicology.
Past-chair, Toxicology and safety evaluation division of Institute of
Food Technology,
Councilor for Food safety subdivision of Society of Toxicology,
Editorial board of Journal of Food Protection
and ad hoc reviewer for numerous journals.
Experience in food toxicology and safety assessment for private
industry, food regulations and FDA compliance issues.
Frequent presentor of food safety issues at national and international
meetings.

www.agnr.umd.edu/AGNRDirectory/Bio.cfm?ID=105509279
Faculty Webpage

Contact Information:
Work phone 301-405-4523
E-mail bmagnuso@...,
Address:
3209 Marie Mount Hall
College Park, MD 20742

Degrees:
Ph.D., Nutrition and Food Science, University of Manitoba
M. Sc, Toxicology, University of Saskatchewan
B.S.H.Ec, Food Science, University of Saskatchewan ]


George A. Burdock b; gburdock@...,

[
www.chpa-info.org/NR/rdonlyres/32E831AF-33CA-4209-B923-19A6332FC052/0/07_11_07_A\
TCH_BurdockCV.pdf
CURRICULUM VITAE

G.A. Burdock, I.G. Carabin and J.C. Griffiths (2007)
Breaking Down the Barriers to Functional Foods, Chapter XX. In:
Nutraceutical And Functional Food Regulation In The United States and
Around The World, a volume of the Foodscience and Technology Series.
D. Bagchi (ed). Elsevier, NY (accepted and in press).

Flavor and Extract Manufactures’ Association (FEMA) 1986-1992
Washington, D.C.
Director of Scientific Affairs
Dr. Burdock managed the FEMA scientific programs, coordinated the
research activities of the testing laboratories, and communicated with
external consultants and allied industry committees working
with FEMA.
As the primary scientific liaison, Dr. Burdock guided member companies
with the preparation of submissions to the FEMA Expert Panel for GRAS
review, alerted Expert Panel and Association members to scientific
developments in the food and flavor industry, and identified
changes in the regulatory policies as a result of these developments.
He authored and edited comprehensive literature reviews on flavor
additives and other topics relevant to the Association’s interests.
EDUCATION
Ph.D. in Toxicology, School of Pharmacy, University of Mississippi, 1980
Master of Combined Sciences, Physiology and Biochemistry, University of
Mississippi, 1973
Bachelor of Science, Biology, University of Mississippi, 1969

F. Kotsonis, and G.A. Burdock (2007) Chapter 30: Food Toxicology.
In: Toxicology: The Basic Science of Poisons. 8th edition C.D. Klaassen
(Ed.) Pergamon Press, New York. (accepted and in press)

http://www.chipsbooks.com/clineval.htm
CLINICAL EVALUATION OF A FOOD ADDITIVE: Assessment of Aspartame
edited by: Christian Tschanz, Harriett Butchko, W.W. Stargel, Frank
Kotsonis 1996 308 pages $134.00 + shipping

http://72.14.253.104/search?q=cache:d6B0UqVkWZIJ:www.reeis.usda.gov/web/areera/R\
eports/2004/AES.WI.pdf+%22Frank+Kotsonis%22,NutraSweet&hl=en&ct=clnk&cd=15&gl=us
Frank Kotsonis –- Dr. Kotsonis was corporate vice president of World
Wide Regulatory Sciences (1995-2000) at the Monsanto Company,
senior vice president of Preclinical and Clinical Research at the
NutraSweet Company,
director of toxicology at G.D.Searle,
and adjunct professor of toxicology at the Philadelphia College of
Pharmacy and Science.
He retired after 23 years at Monsanto in May 2000.

Burdock Group,
888, 17th Street, NW, Suite 810, Washington, DC 20006, USA.

G.A. Burdock, Handbook of Flavour Ingredients, CRC Press, 2002.

G. A. Burdock, Encyclopedia of Food and Color Additives,. Vol. I, CRC,
Boca Raton, FL, 1997. ]


John Doull c; [ Dr. John Doull, MD, PhD
Professor Emeritus
Pharmacology, Toxicology and Therapeutics
Email address: jdoull@...,
Main Phone Number: (913) 588-7508
Mailing Address:
4027 Kansas Life Sciences Innovations Center
Mail Stop 1018
3901 Rainbow Blvd.
Kansas City, KS 66160

Doull's Toxicology: The Basic Science of Poisons (Editors, CD Klaassen,
MO Amdur, J Doull). Fifth Edition.McGraw-Hill, Inc, New York, USA, 1996.

Toxicol Rev. 2005; 24(1): 1-10.
The potential adverse health effects of dental amalgam.
Brownawell AM, Berent S, Brent RL, Bruckner JV, Doull J, Gershwin EM,
Hood RD, Matanoski GM, Rubin R, Weiss B, Karol MH.
"This review has uncovered no convincing evidence pointing to any
adverse health effects that are attributable to dental amalgam
restorations besides hypersensitivity in some individuals.
PMID: 16042501"

Annu Rev Pharmacol Toxicol. 2001; 41: 1-21.
Toxicology comes of age.
Doull J.
Department of Pharmacology, Toxicology, and Therapeutics, University of
Kansas Medical Center, Kansas City, Kansas 66160, USA. jdoull@...

This paper contains recollections of some of the people and events that
influenced the development of toxicology as an academic discipline. It
also describes my experiences in pharmacology at the University of
Chicago and the University of Kansas Medical Center and concludes with
speculation concerning the future of toxicology. Moderation in all
things/Ne quid nimis. --Terence in Andria PMID: 11264448 ]


Robert M. Kroes d; [[deceased] No items in PubMed

1972 -- National Cancer Institute, National Institutes of Health,
Bethesda, Maryland 20014

Contact (for editorial): Bert Brunekreef, Ph.D.,
Institute for Risk Assessment Services, U

(Message over 64k, truncated.)