aspartame induces lymphomas and leukaemias in rats, full plain text, M Soffritti, F Belpoggi, DD Esposti, L Lambertini: Ramazzini Foundation study 2005.07.14: main results agree with their previous methanol and formaldehyde studies: Murray 2005.08.31

************************************************** *************

http://groups.yahoo.com/group/aspartameNM/message/1186
aspartame induces lymphomas and leukaemias in rats, full plain text,
M Soffritti, F Belpoggi, DD Esposti, L Lambertini: Ramazzini Foundation
study 2005.07.14: main results agree with their previous methanol and
formaldehyde studies: Murray 2005.08.31

http://www.ramazzini.it/fondazione/d...ameGEO2005.pdf

" In rodents and humans,
APM is metabolised in the gastrointestinal tract
into three constituents:
aspartic acid, phenylalanine and methanol 3. "

" These experiments demonstrate that the increase in
lymphomas and leukaemias,
observed in the APM study,
could be related to methanol, a metabolite of APM,
which is metabolised to formaldehyde and then to formic acid,
both in humans and rats 3. "

" 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. "


1. The total number of rats was 1800. 1500 were given aspartame.

2. 44 [ 14.7 % ] of the 300 control rats, given no aspartame, developed
lymphomas and leukemias (hemolymphoreticular neoplasias ), and none had
malignant brain tumors.

Of 1500 rats given aspartame, 294 [ 19.6 % ] had lymphomas and leukemias
(hemolymphoreticular neoplasias), and 12 [ 0.8 % ] had malignant brain
tumors.

In their previous methanol study, reported Dec 2002, of 200 + 100 = 300
control rats, given no methanol, there were 41+ 15 = 56 [ 18.7% ]
lymphomas and leukemias (hemolymphoreticular neoplasias), while of 600 +
100 = 700 rats given methanol, there were 187 + 15 = 202 with the same
cancers [ 28.9 % ]. They added 100 rats given 15 ppm methanol to their
Table 3 summarizing the formaldehyde data in their formaldehyde study, in
which their 200 control rats had 15 of these cancers.

In their previous formaldehyde study, reported Dec 2002, 200 control rats,
given no formaldehyde, had 15 [ 7.5 %] lymphomas and leukemias
(hemolymphoreticular neoplasias), while of the 600 rats given formaldehyde,
121 [ 20.3 % ] had these cancers.

Probably, other factors, such as viruses, bacteria, molds, or toxic
chemicals in the air, water, and food, also facilitate these cancers.


http://www.ramazzini.it/eng/fondazio...gli.asp?id=210

News and events
Istituto Ramazzini
Collegium Ramazzini
NEWS AND EVENTS 14 July 2005 Press Release

Results of study on the carcinogenicity of the artificial sweetener
aspartame

CRC/ERF

Results of study on the carcinogenicity of the artificial sweetener
aspartame

Summary.

A long-term study to evaluate the potential carcinogenic effects of
aspartame,
an artificial sweetener used in more than 6,000 food and pharmaceutical
products has recently been completed in the experimental laboratories of its
Cancer Research Center of the European Foundation of Oncology and
Environmental Sciences "B. Ramazzini" in Bologna, Italy.

The first results of the experiment were reported to the Ministry of Health
and to the Superior Institute of Health of the Italian government in April
2005.

In mid-June, these findings were then communicated to
the European Food Safety Authority,
the Herbert Irving Comprehensive Cancer Center of Columbia University,
the National Cancer Institute of the US government,
and the National Toxicology Program of the US National Institutes of Health.

First results demonstrate that aspartame,
when administered to rats for the entire life span,
induces an increase of lymphomas and leukaemias in female rats.

The study is currently being published in the European Journal of Oncology
(available at:
http://www.ramazzini.it/fondazione/d...ameGEO2005.pdf ) and final
results will be presented
at the 3rd international scientific conference of the Collegium Ramazzini,
"Framing the Future in Light of the Past: Living in a Chemical World",
to be held in Bologna, Italy from September 18-21, 2005,
the proceedings of which will be published in
the Annals of the New York Academy of Sciences.

Communication.

Aspartame is an artificial sweetener consumed by hundreds of millions of
people worldwide.

It is used in over 6,000 diet products including soft drinks, chewing gum,
candy, desserts, yogurt as well as in pharmaceuticals, in particular, syrups
and antibiotics for children.

The average daily intake of aspartame is calculated to be about 2-3 mg/Kg of
body weight, a figure which increases for children and women of childbearing
age.

Current daily intake allowed by regulatory bodies is 50 mg/Kg of body weight
in the US and
40 mg/Kg of body weight in the European Union.

Prior to the commercialization of aspartame in the 1970s,
the manufacturers of the compound conducted various experimental studies on
rats and mice to test its carcinogenicity.

When taken together, the results of these studies were considered negative
with regard to the carcinogenicity of aspartame.

Doubts were however raised by some in the scientific community about the
conduct of the experiments and
the fact that some cases of malignant brain tumors were found among animals
treated with aspartame
while none were found among the control group.

Given the limitations of these studies and
the ever growing use of aspartame throughout the years,
the European Ramazzini Foundation decided in the late 1990s
to plan and perform an experiment that would,
based on the total number of animals used,
the number of dose levels studied,
and the conduct of the experiment according to Good Laboratory Practices,
provide an adequate evaluation of the potential carcinogenic effects of
aspartame.

The CRC/ERF study was conducted on 1800 rats (900 males, 900 females)
of the colony used for over 30 years by the Foundation.

In order to simulate daily human intake,
aspartame was added to the standard rat diet in quantities of
5000, 2500, 100, 500, 20, 4, and 0 mg/Kg of body weight.
[ This asserts that humans are twenty times more vulnerable to aspartame
(methanol, formaldehyde, formic acid) toxicity than rats. ]

Treatment of the animals began at 8 weeks of age and
continued until spontaneous death.

A complete necropsy and histopathological evaluation of tissues and organs
was then performed on each deceased animal,
for a total of over 30,000 slides examined by microscope.

The first results of the experiment show:

1) a dose-related statistically significant increase of lymphomas and
leukemias in female rats.
This statistically significant increase was also observed at a dose level of
20 mg/Kg of body weight,
a dose inferior to the accepted daily intake permitted by current
regulations (50-40 mg/Kg of body weight);

2) that the addition of aspartame to the diet induces
a dose-related reduction in food consumption,
without however causing a difference in body weight
between treated and untreated animals.

The above results demonstrate
for the first time
that aspartame is a carcinogenic agent,
capable of inducing lymphomas and leukaemias in female rats,
including when administered at dose levels very close
to the acceptable daily intake for humans.

In addition, the data demonstrate that the integration of aspartame into the
diet did not affect the body weight of treated animals
compared with untreated animals.

As recognized by the International Agency for Research on Cancer (IARC) of
the World Health Organization,
results of long-term bioassays conducted on rodents (rats and mice)
are highly predictive of carcinogenic risk for humans.

In light of this fact,
the results of the CRC/ERF study on aspartame call
for urgent reconsideration of regulations
governing its use as an artificial sweetener
in order to better protect public health,
in particular that of children.

Websites

European Foundation for Oncology and Environmental Sciences "B. Ramazzini"
www.ramazzini.it/fondazione/eng

3rd international scientific conference of the Collegium Ramazzini
www.ramazzini.it/living2005

Contact Kathryn Knowles Director of Resource Development
European Foundation of Oncology and Environmental Sciences "B. Ramazzini"
+39 0516640460

Home | About us | Research and Activities | Publications | News and Events
Support the Foundation | Partners | Contact Us | Credits | Versione Italiana

FONDAZIONE "B. RAMAZZINI" - Via Guerrazzi, 18 - 40125 Bologna - tel. 051
237286 - fax 051 2911679 -

CENTRO DI RICERCA SUL CANCRO - Castello di Bentivoglio, Via Saliceto,
3-40010 Bentivoglio (BO) - tel. 051.66.40.460 - fax 051 6640223

************************************************** *************

http://www.ramazzini.it/fondazione/d...ameGEO2005.pdf

" Conclusions
In our experimental conditions, it has been demonstrated,
for the first time, that APM causes a dose-related
statistically significant increase in lymphomas and leukaemias in females
at dose levels very near
those to which humans can be exposed.

Moreover, it can hardly
be overlooked that at the lowest exposure of 80 ppm,
there was a 62% increase in lymphomas and leukaemias
compared to controls, even though this was not statistically significant.

When compared to the concurrent control
group, an increase in the incidence of these neoplasias
was also observed in males exposed to the highest dose;
even though not statistically significant,
this observation confirms and extends the result in females.

The significance of the increase in haemolymphoreticular
neoplasias is further reinforced by the following considerations,
based on the results of experiments performed in the CRC laboratory.

These experiments demonstrate that the increase in
lymphomas and leukaemias, observed in the APM study,
could be related to methanol,
a metabolite of APM, which
is metabolised to formaldehyde and then to formic acid,
both in humans and rats 3.

In fact we have shown that:

1) methanol administered in drinking water increased the incidence
of lymphomas and leukaemias in female rats 11;

2) the same effect was induced in females treated with the
gasoline oxygenated additive methyl-tert-butyl-ether
(MTBE), which is also metabolised to methanol 12 ;

and finally
3) an increase in the incidence of lymphomas and
leukaemias was also observed in females treated with
formaldehyde 9, 13.

These results further highlight the important role that
formaldehyde has on the induction of haematological
malignancies in rodents.

Moreover, in a recent reevaluation
of the carcinogenicity of formaldehyde by the International
Agency for Research on Cancer (IARC),
strong, although not considered sufficient,
evidence of an association with leukaemias in humans was found 14.

Since the results of carcinogenicity bioassays in rodents,
mainly rats and mice, have been shown to be a consistent
predictor of human cancer risk 15-17,
the first results of our study
call for urgent re-examination of permissible exposure
levels of APM in both food and beverages,
especially to protect children. "

" 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.

10. Harris NL, Jaffe ES, Vardiman JW, et al.
WHO Classification of tumours of haematopoietic and lymphoid tissues:
Introduction.
In
Jaffe ES, Harris NL, Stein H, et al.
Tumours of haematopoietic and lymphoid tissues.
Lyon: IARC Press, 2001, 12-3.

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.

12. Belpoggi F, Soffritti M, Maltoni C.
Methyl-tertiary-butyl ether (MTBE), a gasoline additive, causes testicular
and lymphohaematopoietic cancers in rats.
Toxicol Ind Health 1995; 11: 119-49.

13. Soffritti M, Maltoni C, Maffei F, et al.
Formaldehyde: an experimental multipotent carcinogen.
Toxicol Ind Health 1989; 5: 699-730.

14. International Agency for Research on Cancer.
Monographs on the evaluation of the carcinogenic risk of chemicals to
humans.
Formaldehyde, 2-Butoxyethanol and 1-tert-Butoxy-2-Propanol.
Vol. 88 (in press). Available on http://www.iarc.fr.

15. Huff J. Long-term chemical carcinogenesis bioassays predict
human cancer hazards. Issues, controversies, and uncertainties.
In
Bailer JA, Maltoni C, Bailar III JC, et al.
Uncertainty in the risk assessment of environmental and occupational
hazards.
Ann NY Acad Sci 1999; 895: 56-79.

16. Tomatis L, Aitio A, Wilbourn J, et al.
Human carcinogens so far identified.
Jpn J Cancer Res 1989; 80: 795-807.

17. Rall DP.
Can laboratory animal carcinogenicity studies predict cancer in exposed
children?
Environ Health Perspect 1995; 103 suppl 6: 173-5. "


http://www.ramazzini.it/fondazione/d...ameGEO2005.pdf

page 1

Original studies/Studi originali
General topics/Argomenti generali

Eur. J. Oncol., vol. 10, n. 2, pp. 00-00, 2005 IN PRESS

Aspartame induces lymphomas and leukaemias in rats *
L'aspartame induce linfomi e leucemie nei ratti
Morando Soffritti,
Fiorella Belpoggi,
Davide Degli Esposti,
Luca Lambertini

Cancer Research Centre, European Ramazzini Foundation of Oncology and
Environmental Sciences, Bologna, Italy
Received/Pervenuto 15.3.2005 - Accepted/Accettato 11.4.2005

Address/Indirizzo: Dr. Morando Soffritti, Centro di Ricerca sul Cancro,
Fondazione Europea di Oncologia e Scienze Ambientali "B. Ramazzini",
Castello di Bentivoglio, 40010 Bentivoglio (BO), Italia - Tel.
+39/051/6640460 - Fax +39/051/6640223 - E-mail:

* Research supported by European Ramazzini Foundation of Oncology and
Environmental Sciences, Bologna, Italy


Summary

Aspartame, a widely used artificial sweetener, was administered
with feed to male and female Sprague-Dawley rats
(100-150/sex/group),
8 weeks-old at the start of the experiment,
at concentrations of 100,000; 50,000; 10,000; 2,000; 400; 80 and 0 ppm.
Treatment lasted until spontaneous death of the animals.

In this report we present the first results showing that aspartame,
in our experimental conditions,
causes a statistically significant, dose-related increase
in lymphomas and leukaemias in females.

No statistically significant increase in malignant brain tumours was
observed among animals from the treated groups
as compared to controls.
Eur. J. Oncol., 10 (2), 00-00, 2005

Key words: aspartame, artificial sweetener, carcinogenesis,
rats, lymphoma, leukaemia

Riassunto

L'aspartame, un dolcificante artificiale largamente
diffuso, è stato somministrato con il mangime a ratti
Sprague-Dawley, maschi e femmine (100-150/sesso/
gruppo), di 8 settimane di età all'inizio dell'esperimento,
a concentrazioni di 100.000; 50.000; 10.000;
2.000; 400; 80 e 0 ppm.
Il trattamento è durato fino alla
morte spontanea degli animali. In questo articolo
vengono presentati i primi risultati che dimostrano
come l'aspartame, nelle nostre condizioni sperimentali,
causa un incremento statisticamente significativo,
dose-correlato, di linfomi e leucemie nelle femmine.
Nei gruppi trattati rispetto al controllo non è stato osservato
nessun aumento statisticamente significativo
dei tumori maligni del cervello.
Eur. J. Oncol., 10 (2), 00-00, 2005

Parole chiave: aspartame, dolcificante artificiale, cancerogenesi,
ratti, linfoma, leucemia

Introduction

Aspartame (APM) is a widely used artificial sweetener
consumed by hundreds of millions of people around
the world 1, 2.

It is found in more than 6,000 products, including
soft drinks, chewing gum, candy, yoghurt, tabletop
sweeteners and some pharmaceuticals such as vitamins
and sugar-free cough drops 2.

Dietary surveys, performed among APM consumers,
have shown that the average APM daily intake in the general
population ranged from 2 to 3 mg/kg b.w.
and was even more in children and pregnant women 1.

The Acceptable Daily Intake (ADI) both in the US and in Europe is
50 and 40 mg/kg b.w., respectively 1.

In rodents and humans,
APM is metabolised in the gastrointestinal tract
into three constituents:
aspartic acid, phenylalanine and methanol 3.

Three long-term feeding carcinogenicity bioassays on
APM were performed on rats, and one on mice, during the 1970s.

Overall, the carcinogenicity studies were considered negative 4,
but it must be noted that these studies did not comply
with the basic requirements which must nowadays be met when testing the
carcinogenicity potential of a chemical or physical agent.

Because of these limitations,
we decided to perform a mega-experiment following
the currently accepted Good Laboratory Practices.

In the present paper we are reporting our first results on
the incidence of haemolymphoreticular malignancies
(lymphomas and leukaemias) and malignant brain tumours.

Materials and methods

The APM used was produced by Nutrasweet and supplied
by Giusto Faravelli S.p.A., Milan, Italy.

As an active ingredient, its purity was more than 98%.

To simulate an assumed daily intake by humans of
5,000; 2,500; 500; 100; 20; 4; or 0 mg/kg b.w.,
APM was added to the standard Corticella diet,
used for 30 years at the laboratory of
the Cancer Research Centre (CRC) of the European Ramazzini Foundation (ERF),
at concentrations of
100,000; 50,000; 10,000; 2,000; 400; 80; or 0 ppm.
[ This asserts that humans are twenty times more vulnerable to aspartame
(methanol, formaldehyde, formic acid) toxicity than rats. ]

APM-treated feed was administered ad libitum to Sprague-Dawley rats
(100-150/sex/group),
8 weeks old at the start of the experiment,
and the treatment lasted until spontaneous death.

Control animals received the same feed without APM.
The plan of the experiment is shown in Table 1.

page 2 M. Soffritti, F. Belpoggi, D. Degli Esposti, et al. IN PRESS

Table 1 - Long-term carcinogenicity bioassay on aspartame administered with
feed supplied ad libitum to male (M) and female (F) Sprague-Dawley rats from
8 weeks of age until spontaneous death.

Plan of the experiment.
Age at start of treatment was 8 weeks. Duration was then natural life span.
Group Sex (M, F, M+F) Animals (No) Treatment Dose (ppm) mg/kg b.w. a Human
ADI equivalent b (X)

I
---M 100---------100,000-------5,000------100X
---F 100
M+F 200

II
---M 100----------50,000-------2,500-------50X
---F 100
M+F 200

III
---M 100----------10,000---------500-------10X
---F 100
M+F 200

IV
---M 150-----------2,000---------100--------2X
---F 150
M+F 300

V
---M 150-------------400----------20--------0.4X
---F 150
M+F 300

VI
---M 150--------------80-----------4---------0.08X
---F 150
M+F 300

VII
---M 150---------------0------------0---------0
---F 150
M+F 300

a The daily assumption in mg/kg b.w. was calculated considering the average
weight of a rat for the duration of the experiment as 400 g,
and the average consumption of feed as 20 g per day, both for males and
females

b Considering the Acceptable Daily Intake (ADI) of 50 mg/kg b.w. for humans
[ This asserts that humans are twenty times more vulnerable to aspartame
(methanol, formaldehyde, formic acid) toxicity than rats. ]


Male (M) and female (F) rats from the colony of the CRC were used.
This colony of rats has been employed
for various experiments in the CRC Laboratory for nearly 30 years.
Data are available on the tumour incidence among untreated Sprague-Dawley
rats.
These animals were monitored for feed, water consumption, and body
weight, for their life span and, at death, underwent complete necropsy and
histopathological evaluation (historical controls).

The experiment was conducted according to the Italian law regulating use of
animals for scientific purposes 5.

After weaning, at 4-5 weeks of age,
the experimental animals were identified by ear punch,
randomised in order to have no more than one male and one female from
each litter in the same group, and housed in groups of 5 in makrolon cages
(41x25x15 cm), with stainless-steel wire tops and a shallow layer of white
wood shavings as bedding. The animals were kept in one single room, at 23
±2°C and 50-60% relative humidity.

Once a week for the first 13 weeks, then every two weeks until 110 weeks of
age, the mean daily drinking water and feed consumption were measured per
cage, and body weight individually.
Body weight continued to be measured every 8 weeks until the end of the
experiment.

Status and behaviour of the animals were examined 3 times daily, and
they were clinically examined for gross changes every 2 weeks.
All animals were kept under observation until spontaneous death.

page 3 Aspartame, a leukaemogenic compound IN PRESS


Fig. 1. Mean daily water consumption in male Sprague-Dawley rats

Fig. 2. Mean daily water consumption in female Sprague-Dawley rats


The biophase of the experiment terminated after 151 weeks,
with the death of the last animal at the age of 159 weeks.

Upon death, the animals underwent complete necropsy.
Histopathology was routinely performed on the following
organs and tissues of all animals from each group:
skin and subcutaneous tissue, mammary gland, the brain (3 sagittal
sections), pituitary gland, Zymbal glands, salivary glands, Harderian
glands, cranium (five sections, with oral and nasal cavities and external
and internal ear ducts), tongue, thyroid, parathyroid, pharynx, larynx,
thymus and mediastinal lymph nodes, trachea, lung and mainstem bronchi,
heart, diaphragm, liver, spleen, pancreas, kidneys, adrenal glands,
oesophagus, stomach (fore and glandular), intestine (four levels), urinary
bladder, prostate, gonads, interscapular brown fat pad, subcutaneous
and mesenteric lymph nodes and other organs or tissues with pathological
lesions.

All organs and tissues were preserved in 70% ethyl alcohol, except for bones
which were fixed in 10% formalin and then decalcified with 10% formaldehyde
and 20% formic acid in water solution.
The normal specimens were trimmed,
following the Standard Operating Procedures at the CRC Laboratory: i.e.
parenchymal organs were dissected through the hilus to expose the widest
surface, and hollow organs were sectioned across the greatest diameter.

page 4 M. Soffritti, F. Belpoggi, D. Degli Esposti, et al. IN PRESS


Fig. 3. Mean daily feed consumption in male Sprague-Dawley rats

Fig. 4. Mean daily feed consumption in female Sprague-Dawley rats


Any pathological tissue was trimmed through the largest surface, including
normal adjacent tissue. Trimmed specimens were processed as paraffin blocks,
and 3-5 micron sections of every specimen were obtained.
Sections were routinely stained with haematoxylin-eosin.

Statistical analyses were performed using the poly-k test (k = 3). This test
is a survival-adjusted quantal-response procedure that modifies the
Cochran-Armitage linear trend test to take survival differences into account
6-8.

Results

During the experiment no differences were observed among the various groups
in mean daily water consumption (figs. 1 and 2).

A dose-related difference in feed consumption was observed between the
various treated groups and the control group in both males and females
(figs. 3 and 4).

No differences in mean body weight were observed among treated and control
groups in either males or females (figs. 5 and 6).

No substantial difference in survival was observed among treated and control
groups, males or females (figs. 7 and 8).

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.

The occurrence of lymphomas and leukaemias among male and female rats in
treated and control groups is shown in Table 2.

page 5 Aspartame, a leukaemogenic compound IN PRESS


Fig. 5. Mean body weights in male Sprague-Dawley rats

Fig. 6. Mean body weights in female Sprague-Dawley rats


The data indicate that APM causes a statistically significant increase in
the incidence of lymphomas and leukaemias in females, at concentrations of
100,000 (p= 0.01);
50,000 (p= 0.01);
10,000 (p= 0.05);
2,000 (p= 0.01) and
400 (p= 0.01) ppm
as compared to untreated controls.
This increase is dose-related (p= 0.05).

Although not statistically significant,
an increase was also observed in females treated with 80 ppm and
in males treated with the highest dose.

The haemolymphoreticular neoplasias observed in the experiment include:
lymphoblastic lymphoma and leukaemia,
lymphocytic lymphoma,
lymphoimmunoblastic lymphoma,
histiocytic sarcoma and monocytic leukaemia,
myeloid leukaemia.

The most frequent type of neoplasia was the lymphoimmunoblastic lymphoma
(figs. 9 and 10).

Lymphomas and leukaemias are considered together,
since both solid and circulating phases are present in many lymphoid
neoplasms, and distinction between them is artificial 10.

The occurrence of brain malignancies is shown in Table 3.

Sparse malignant brain tumours were observed
among males and females in the treated groups and
none in the controls.

page 6 M. Soffritti, F. Belpoggi, D. Degli Esposti, et al. IN PRESS


Fig. 7. Survival in male Sprague-Dawley rats

Fig. 8. Survival in female Sprague-Dawley rats


page 7 Aspartame, a leukaemogenic compound IN PRESS


Fig. 9. Lymphoimmunoblastic lymphoma in a female rat administered
80 ppm aspartame in feed (lung). HE X 25

Fig. 10. A detail of the lymphoimmunoblastic lymphoma shown in fig. 9.
HE X 400


In our historical controls over the last 20 years,
when we consider groups of 100 or more animals per sex
(1934 males and 1957 females),the overall incidence of lymphomas and
leukaemias in males is 21.8% (8.0-30.9) and
in females 13.4% (7.0-18.4) .


The overall incidence of malignant brain tumours is
1.7% (0-5.0) in males and
0.7% (0-2.0) in females respectively.

page 8 M. Soffritti, F. Belpoggi, D. Degli Esposti, et al. IN PRESS


Table 2 - Long-term carcinogenicity bioassay on aspartame administered with
feed supplied ad libitum to male (M) and female (F) Sprague-Dawley rats from
8 weeks of age until spontaneous death.
Incidence of lymphomas and leukaemias


Table 3 - Long-term carcinogenicity bioassay on aspartame administered with
feed supplied ad libitum to male (M) and female (F) Sprague-Dawley rats from
8 weeks of age until spontaneous death. Incidence of malignant brain tumors
b


Group (100-150 rats each) Dose (ppm) Animals with lymphomas and leukaemias
Animals with malignant brain tumours (No, %, No, %) @

I-----100,000-ppm-29--------29.0-------1-------1.0
---------------------25--------25.0**----1-------1.0
---------------------54--------27.0-------2-------1.0

II-----50,000-ppm-20--------20.0-------2-------2.0
---------------------25--------25.0**----1-------1.0
---------------------45--------22.5-------3-------1.5

III----10,000-ppm-15--------15.0-------0--------0
--------------------19--------19.0*------1--------1.0
--------------------34--------17.0-------1--------0.5

IV------2,000-ppm-33-------22.0-------2--------1.3
---------------------28-------18.7*------1--------0.7
---------------------61-------20.3-------3--------1.0

V---------400-ppm-25-------16.7-------0--------0
---------------------30-------20.0**-----0--------0
---------------------55-------18.3--------0--------0

VI---------80-ppm-23 -------15.3-------2-------1.3
---------------------22 -------14.7-------1-------0.7
---------------------45-------15.0--------3-------1.0

VII---------0-ppm-31--------20.7-------0-------0 [control groups]
--------------------13---------8.7--------0-------0
--------------------44--------14.7--------0-------0

@ The malignancies observed we
10 malignant gliomas or mixed gliomas,
1 medulloblastoma, and
1 malignant meningioma

* Statistically significant p= 0.05;
** Statistically significant p= 0.01 using poly-k test (k = 3)


Conclusions

In our experimental conditions,
it has been demonstrated,
for the first time,
that APM causes a dose-related statistically significant increase
in lymphomas and leukaemias in females
at dose levels very near
those to which humans can be exposed.

Moreover, it can hardly
be overlooked that at the lowest exposure of 80 ppm,
there was a 62% increase in lymphomas and leukaemias
compared to controls,
even though this was not statistically significant.

When compared to the concurrent control group,
an increase in the incidence of these neoplasias
was also observed in males exposed to the highest dose;
even though not statistically significant, this observation
confirms and extends the result in females.

The significance of the increase in haemolymphoreticular neoplasias
is further reinforced by the following considerations,
based on the results of experiments performed in the CRC laboratory.

These experiments demonstrate that the increase in
lymphomas and leukaemias,
observed in the APM study,
could be related to methanol, a metabolite of APM,
which is metabolised to formaldehyde and then to formic acid,
both in humans and rats 3.

In fact we have shown that:

1) methanol administered in drinking water increased
the incidence of lymphomas and leukaemias in female rats 11;

2) the same effect was induced in females treated with the
gasoline oxygenated additive methyl-tert-butyl-ether
(MTBE), which is also metabolised to methanol 12;

and finally
3) an increase in the incidence of lymphomas and leukaemias was also
observed in females treated with formaldehyde 9, 13.

These results further highlight the important role that formaldehyde has on
the induction of haematological malignancies in rodents.

Moreover, in a recent reevaluation of the carcinogenicity of formaldehyde by
the International Agency for Research on Cancer (IARC), strong,
although not considered sufficient,
evidence of an association with leukaemias in humans was found 14.

Since the results of carcinogenicity bioassays in rodents,
mainly rats and mice, have been shown to be
a consistent predictor of human cancer risk 15-17,
the first results of our study call for urgent re-examination
of permissible exposure levels of APM in both food and beverages,
especially to protect children.

References

1. Butchko HH, Stargel WW, Comer CP, et al.
Preclinical safety evaluation of aspartame.
Regul Toxicol Pharmacol 2002; 35: S7-S12.

2. Aspartame Information Center.
Available on http://www.aspartame.org, 2004.

3. Ranney RE, Opperman JA, Maldoon E, et al.
Comparative metabolism of aspartame in experimental animals and humans.
Toxicol Environ Health 1976; 2: 441-51.

4. Food and Drug Administration. Aspartame:
Commissioner's Final Decision; 1981 Fed Regist 46, 38285-308.

5. Repubblica Italiana.
Decreto Legislativo 116.
Attuazione della direttiva n. 86/609/CEE in materia di protezione degli
animali utilizzati a fini sperimentali o ad altri fini scientifici.
Supplemento ordinario alla Gazzetta Ufficiale 1992; 40: 5-25.

page 9 Aspartame, a leukaemogenic compound IN PRESS

6. Bailer AJ, Portier CJ.
Effects of treatment-induced mortality and tumor-induced mortality on tests
for carcinogenicity in small samples.
Biometrics 1988; 44: 417-31.

7. Portier CJ, Bailer AJ.
Testing for increased carcinogenicity using a survival-adjusted quantal
response test. Fundam Appl Toxicol 1989; 12: 731-7.

8. Piergorsh WW, Bailer AJ.
Statistics for environmental biology and toxicology.
London: Chapman, 1997.

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.

10. Harris NL, Jaffe ES, Vardiman JW, et al.
WHO Classification of tumours of haematopoietic and lymphoid tissues:
Introduction.
In
Jaffe ES, Harris NL, Stein H, et al.
Tumours of haematopoietic and lymphoid tissues. Lyon: IARC Press, 2001,
12-3.

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.

12. Belpoggi F, Soffritti M, Maltoni C.
Methyl-tertiary-butyl ether (MTBE), a gasoline additive, causes testicular
and lymphohaematopoietic cancers in rats.
Toxicol Ind Health 1995; 11: 119-49.

13. Soffritti M, Maltoni C, Maffei F, et al.
Formaldehyde: an experimental multipotent carcinogen.
Toxicol Ind Health 1989; 5: 699-730.

14. International Agency for Research on Cancer.
Monographs on the evaluation of the carcinogenic risk of chemicals to
humans. Formaldehyde, 2-Butoxyethanol and 1-tert-Butoxy-2-Propanol.
Vol. 88 (in press). Available on http://www.iarc.fr.

15. Huff J.
Long-term chemical carcinogenesis bioassays predict human cancer hazards.
Issues, controversies, and uncertainties.
In
Bailer JA, Maltoni C, Bailar III JC, et al.
Uncertainty in the risk assessment of environmental and occupational
hazards.
Ann NY Acad Sci 1999; 895: 56-79.

16. Tomatis L, Aitio A, Wilbourn J, et al.
Human carcinogens so far identified.
Jpn J Cancer Res 1989; 80: 795-807.

17. Rall DP.
Can laboratory animal carcinogenicity studies predict cancer in exposed
children?
Environ Health Perspect 1995; 103 suppl 6: 173-5.

page 10 M. Soffritti, F. Belpoggi, D. Degli Esposti, et al. IN PRESS
************************************************** *************

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.

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:

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

J. Nutrition 1973 Oct; 103(10): 1454-1459.
Metabolism of aspartame in monkeys.
Oppermann JA, Muldoon E, Ranney RE.
Dept. of Biochemistry, Searle Laboratories,
Division of G.D. Searle and Co. Box 5110, Chicago, IL 60680
They found that about 70% of the radioactive methanol in aspartame put into
the stomachs of 3 to 7 kg monkeys was eliminated within 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.


Males
Females
Males + Females

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

Group
100 rats each

aspartame dose a
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

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

a 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, p0.05 using X2 test
^ methanol, p0.05 using Cochrane-Armitage test for dose-response
relationship

* formaldemyde, p0.05 using X2 test
** formaldehyde, p0.01 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:

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

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/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 rmforall


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

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.
************************************************** *************

Rich Murray, MA Room For All 505-501-2298
1943 Otowi Road Santa Fe, New Mexico 87505 USA
http://groups.yahoo.com/group/aspartameNM/messages
group with 146 members, 1,208 posts in a public, searchable archive

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

http://groups.yahoo.com/group/aspartameNM/message/1185
Ramazzini Institute (Italy) lifetime study with 1800 rats shows aspartame at
human use levels causes cancer (methanol, formaldehyde, formic acid), M
Soffritti and F Belpoggi: Felicity Lawrence, The Guardian (UK): Murray
2005.07.15

http://groups.yahoo.com/group/aspartameNM/message/1189
Michael F Jacobson of CSPI now and in 1985 re aspartame toxicity, letter to
FDA Commissioner Lester Crawford; California OEHHA aspartame critique
2004.03.12; Center for Consumer Freedom denounces CSPI: Murray 2004.07.27

http://groups.yahoo.com/group/aspartameNM/message/1045
http://www.holisticmed.com/aspartame...2-response.htm
Mark Gold exhaustively critiques European Commission Scientific
Committee on Food re aspartame ( 2002.12.04 ): 59 pages, 230 references

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

Gold points out that industry methanol assays were too insensitive to
properly measure blood methanol levels. ]

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 is 18
times the USA EPA limit for daily formaldehyde in drinking water, 2 mg in 2
L water.

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

Aspartame is made of phenylalanine (50% by weight) and aspartic acid (39%),
both ordinary amino acids, bound loosely together by methanol (wood alcohol,
11%). The readily released methanol from aspartame is within hours turned
by the liver into formaldehyde and then formic acid, both potent, cumulative
toxins.


http://groups.yahoo.com/group/aspartameNM/message/1182
Joining together: short review: research on aspartame (methanol,
formaldehyde, formic acid) toxicity: Murray 2005.07.08 rmforall

http://groups.yahoo.com/group/aspartameNM/message/1071
research on aspartame (methanol, formaldehyde, formic acid) toxicity: Murray
2004.04.29 rmforall

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

http://groups.yahoo.com/group/aspartameNM/message/1131
genotoxicity of aspartame in human lymphocytes 2004.07.29 full plain text,
Rencuzogullari E et al, Cukurova University, Adana, Turkey 2004 Aug: Murray
2004.11.06 rmforall

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

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 rmforall

Thrasher (2001): "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 ]

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

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

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 rmforall


Clearly, Europe has placed the issue of aspartame toxicity on the table
as a legitimate, urgent issue for evidence-based public discussion.

Perhaps this shift in the climate of opinion is due to:
European Ramazzini Foundation, led by Morando Soffritti, MD.
Cancer Research Center, European Ramazzini Foundation for
Oncology and Environmental Sciences, Bentivoglio Castle, 40010
Bentivoglio (BO), Italy. +39-051-6640460 fax +39-051-6640223

Annuals of the New York Academy of Science. 2002 Dec; Vol. 982:

The RF research program was started in 1966 by the eminent Cesare
Maltoni, (1930-2001):
p. 6 "Maltoni was known for his meticulous and carefully documented
experiments.
He studied 198 chemicals and agents and conducted 394 separate experiments
using 138,281 animals.
Of the 135 agents studied,
68.9% were found to be carcinogenic,
5.92% showed borderline carcinogenicity,
and 25.18% were found to be noncarcinogenic in the animals tested."
Often, the hundreds to thousands of rats in each study were exposed daily
for two years and then thoroughly examined for cancers after their later
natural deaths.

http://www.nyas.org/books/vols/v982.html
Annals of the New York Academy of Sciences
Carcinogenesis Bioassays and Protecting Public Health:
Commemorating the Lifework of Cesare Maltoni and Colleagues
Edited by Myron A. Mehlman (Collegium Ramazzini, Princeton, NJ);
[Dept. of Environmental Medicine, The Mount Sinai Medical Center, New
York City, 609-683-4750]
Eula Bingham (University of Cincinnati College of Medicine, Cincinnati, OH);
Philip J. Landrigan (Mount Sinai School of Medicine, New York, NY);
Morando Soffritti, Fiorella Belpoggi, European Ramazzini Foundation;
Ronald L. Melnick, National Institute of Environmental Health Sciences,
Research Triangle Park, NC

Proceedings of an April 29-30, 2002 Academy conference.
Volume 982 ISBN 1-57331-406-4
231 pages 14 papers 0 posters Price: $135.00
Member Price *: $15.00 December 2002

Long-term experimental carcinogenesis studies are the cornerstone of
human health protection and risk assessment for drugs and chemicals.
Great contributions to the development of bioassay methodology and the
understanding of the mechanisms of carcinogenesis were made by Professor
Cesare Maltoni at the European Foundation of Oncology and Environmental
Sciences "B. Ramazzini," Bologna, Italy.
This volume is based on a conference that was held on the first anniversary
of Professor Maltoni's death to honor him and to celebrate the work on
carcinogenesis bioassays carried out at the Ramazzini Foundation Cancer
Research Center in Italy and at the National Toxicology Program, NIEHS, in
the United States.
Papers include reviews of previously unreported findings and discussion
of the continued utility of such studies for the protection of public
health.
Full text of volume 837 and forward is available at Annals
Online to Academy Members at Members Online and to subscribing
libraries. New York Academy of Sciences 2 East 63rd St., NY, NY 10021

"(3) formaldehyde may produce lymphomas and leukemias..."

Ann N Y Acad Sci. 2002 Dec; 982: 26-45.
Ramazzini Foundation cancer program: history and major projects,
life-span carcinogenicity bioassay design, chemicals studied, and results.
Soffritti M, Belpoggi F, Minardi F, Maltoni C.
Cancer Research Center, European Ramazzini Foundation for Oncology and
Environmental Sciences, Bologna, Italy.

The Ramazzini Foundation research program was started over thirty years
ago. The features of this program a
(1) systematic and integrated project design;
(2) consistency over time;
(3) homogeneity of approach: key members of the team remain unchanged;
and (4) choice to work on new frontiers of scientific research.
The program centers mainly on three projects:
Project 1: experimental carcinogenicity bioassays;
Project 2: experimental anticarcinogenesis assays to identify factors
and active principles (compounds) capable of opposing the onset of
tumors while being suitable for preventive/chemopreventive intervention;
Project 3: epidemiological studies, both descriptive and analytical, on
tumor incidence and mortality in persons professionally and
environmentally exposed to industrial carcinogenic risks.
The project involving experimental carcinogenicity bioassays for the
identification of exogenous carcinogens (environmental and industrial
above all) began in 1966.
This project has included 398 experimental bioassays on 200
compounds/agents using some 148,000 animals monitored until their
spontaneous death.
Among the studies already concluded, 47 agents have shown "clear
evidence" of carcinogenicity.
The results have demonstrated for the first time that
(1) vinyl chloride can cause liver angiosarcoma as well as other tumors;
(2) benzene is carcinogenic in experimental animals for various tissues
and organs;
(3) formaldehyde may produce lymphomas and leukemias; and
(4) methyl-tert-butyl ether (MTBE), the most common oxygenated additive
used in gasolines, can cause lymphomas/leukemias.
Many of the results achieved have led to the introduction of norms and
measures of primary prevention.
Publication Types: Historical Article PMID: 12562627


p. 48 "The sweetening agent aspartame hydrolyzes in the gastrointestinal
tract to become free methyl alcohol. (25)"
"(25) Medinsky MA & Dorman DC. 1994; Assessing risks of low-level
methanol exposure. CIIT Act. 14: 1-7.
(30) Monte WC. 1984; Aspartame, methanol and the public health.
Journal Applied Nutrition. Vol 36: 42-54."

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.

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.
[ Since 11% of aspartame is its methanol component, always quickly released
into the GI tract upon ingestion, the above methanol levels correspond to
about
200,000, 50,000, 5,000, and 0 ppm aspartame levels,
while the 2005 aspartame levels used were 100,000, 50,000, 10,000, 2,000,
400, 80, and 0 ppm.

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


p. 88 "The sweetening agent aspartame hydrolyzes in the gastrointestinal
tract to become free methyl alcohol, which is metabolized in the liver
to formaldehyde, formic acid, and CO2. (11) [Medinsky & Dorman 1994]"

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.
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

Surely the authors deliberately emphasized that aspartame is well-known
to be a source of formaldehyde, which is an extremely potent, cumulative
toxin, with complex, multiple effects on all tissues and organs.

This is even more significant, considering that they have already tested
aspartame, but not yet released the results: [ comment made spring, 2003 ]

p. 29-32 Table 1: The Ramazzinni Foundation Cancer Program
Project of [200] Long-Term Carcinogenicity Bioassays: Agents Studied

No. No. of Bioassays Species No. Route of Exposure
108. "Coca-Cola" 4 Rat 1,999 Ingestion, Transplantal Route
109. "Pepsi-Cola" 1 Rat 400 Ingestion
110. Sucrose 1 Rat 400 Ingestion
111. Caffeine 1 Rat 800 Ingestion
112. Aspartame 1 Rat 1,800 Ingestion

http://members.nyas.org/events/confe...f_02_0429.html
Soffritti said that Coca-Cola showed no carcinogenicity.
************************************************** ***********

http://www.annalsnyas.org/content/vol982/issue1/ Table of Contents
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.
Cancer Research Center, European Ramazzini Foundation for Oncology and
Environmental Sciences, Bologna, Italy.
MORANDO SOFFRITTI, FIORELLA BELPOGGI, LUCA LAMBERTIN, MICHELINA LAURIOLA,
MICHELA PADOVANI, and CESARE MALTONI

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.
MORANDO SOFFRITTI, FIORELLA BELPOGGI, DANIELA CEVOLANI, MARINA GUARINO,
MICHELA PADOVANI, and CESARE MALTONI

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),
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


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/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 rmforall

http://www.epa.gov/iris/subst/0305.htm
also http://www.china-pops.net/enwww/IRIS...subst/0305.htm 1998.05.05

USA Environmental Protection Agency EPA
Integrated Risk Information System IRIS

This site explains that the harmful rat dose of 500 mg/kg body weight per
day was divided
by 10 for "interspecies extrapolation" (the higher vulnerability of
humans than rats),
by 10 for "range of sensitivity" (the variation of individual human
vulnerability), and
by 10 for "subchronic to chronic exposure" (the increased danger from
lifetime as compared to the 3 month exposure in the rat test),
giving a total reduction of 10x10x10 = 1000 for the UF = Uncertainty Factor.

The human Oral RfD is the rat Oral RfD divided by 1000, so
500 mg/kg/day is reduced to 0.5 mg/kg/day , so that the allowed dose for a
60 kg human is 30 mg oral methanol daily.

Moreover, a recent study found that after 4 months of moderate oral
aspartame, 12 rats took four times longer to finish a simple, one-turn
maze-- an alarming level of neurotoxicity:

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

"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) 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."

[ The 2005 Ramazzini study found that 2 years ingested aspartame at
20, 100, 500, 2,500, and 5,000 mg/kg levels all produced a significant,
substantial increase in female rats of lymphomas and leukaemias by the time
of natural death. ]

The 11% methanol component of aspartame is immediately released in the GI
tract, so these rats were being exposed to only 27.5 mg/kg/day methanol.

The EPA IRIS on 1998.05.05 used a 1986 90 day rat study to find a
No-Observed-Effect Level (NOEL) value of 500 mg/kg/day, which, divided by
1000, became their human long-term safe methanol level of 0.5 mg per kg body
weight per day, which for a 60 kg average person is 30 mg methanol daily,
for oral exposure.

However, the rat level is 18 times greater than that for the level of
dramatic memory loss and clear-cut brain changes found by McConnaughey M,
May 2004.

This suggests reducing the human long-term safe level twenty times to
..025 mg/kg/day = 25 micrograms per kg body weight per day,
which for a 60 kg average person is 1.5 mg oral methanol per day.

Since methanol from any source, once in the human blood stream, is always
quickly and largely turned into formaldehyde and then formic acid, resulting
in durable retained cumulative complex toxic products, this implies a
somewhat smaller formaldehyde ingestion limit. A third of the methanol
would lead to a limit of 0.5 mg ingested and inhaled formaldehyde daily for
a 60 kg average person.


http://groups.yahoo.com/group/aspartameNM/message/1141
Nurses Health Study can quickly reveal the extent of aspartame (methanol,
formaldehyde, formic acid) toxicity: Murray 2004.11.21 rmforall

The Nurses Health Study is a bonanza of information about the health of
probably hundreds of nurses who use 6 or more cans daily of diet soft
drinks -- they have also stored blood and tissue samples from their immense
pool of subjects.

Dark wines and liquors, as well as aspartame, provide similar levels of
methanol, above 100 mg daily, for long-term heavy users. Methanol is
inevitably largely turned into formaldehyde, and thence largely into formic
acid.

Both products are toxic, and at this level of use, about 2 L daily,
almost six 12-oz cans of diet drink, are above recent lifetime EPA
safety limits in tap water for methanol and formaldehyde of respectively,
for a 60 kg person, 30 mg and 9 mg daily. The 1999 EPA level for
formaldehyde in drinking water was 1 ppm, while recent WHO levels are 2.6
ppm.

The immediate health effects for dark wines and liquors are the infamous
"morning after" hangover, for which many informed experts cite as the major
cause the conversion of the methanol impurity, over one part in ten thousand
(red wine has 128 mg/L methanol), into formaldehyde and formic acid.
Everyone knows the complex progression of symptoms at this level of
long-term, chronic toxicity.

Aspartame reactors have a very similar progression.

If 1% of all people exposed to aspartame are heavy users with symptoms, then
there would easily be about 2 million cases in the USA alone.

This is a public health emergency.

At the very least, professionals and the public should be alerted to
investigate their own exposure, and be given a chance to try a very safe,
simple, inexpensive treatment for complex, intractable, progressive
symptoms -- reducing or eliminating their intake.

There are as well, many safe substances that prevent or treat the
toxicities -- for example, high folic acid levels expedite the elimination
of formaldehyde.

These toxicities are largely uncontrolled co-factors that affect every
disease and must confuse and impede many health research programs on all
levels.

People in high-pressure, critical occupations, such as pilots, nuclear plant
operators, and national leaders, should certainly be alerted.

Also, two careful studies show substantial methanol release from degradation
of pectins by bacteria in the colon from fruits and vegetables -- a topic
that deserves careful, thorough research.

Due to my bias, based on detailed reviews by Monte WC (1984)
and by Mark D Gold (2003), for months I have been discounting the
startlingly high methanol levels reported in the abstract for Lindinger W
(1997). I had been reducing the values in their abstract from g to mg, an
unwarrented "correction" by a factor of a thousand, only to find that
thefull text study and their many related studies supply expert, robust
results:

Alcohol Clin Exp Res. 1997 Aug; 21(5): 939-43.
Endogenous production of methanol after the consumption of fruit.
Lindinger W, Taucher J, Jordan A, Hansel A, Vogel W.
Institut fur Ionenphysik, Leopold Franzens Universitat Innsbruck, Austria.

After the consumption of fruit, the concentration of methanol in the human
body increases by as much as an order of magnitude.
This is due to the degradation of natural pectin (which is esterified with
methyl alcohol) in the human colon.
In vivo tests performed by means of proton-transfer-reaction mass
spectrometry show that consumed pectin in either a pure form (10 to 15 g)
or a natural form (in 1 kg of apples) induces a significant increase of
methanol in the breath (and by inference in the blood) of humans.
The amount generated from pectin (0.4 to 1.4 g) [ 400 to 1400 mg ]
is approximately equivalent to the total daily endogenous production
(measured to be 0.3 to 0.6 g/day) [ 300 to 600 mg ]
or that obtained from 0.3 liters of 80-proof brandy
(calculated to be 0.5 g). [ 500 mg ]
This dietary pectin may contribute to the development
of nonalcoholic cirrhosis of the liver. PMID: 9267548

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.
PMID: 8561283 [ Corrected 2005.07.11:
24 ml means 19 g ethanol, and 0.19 ml means 0.15 g = 150 mg methanol.
One L diet soda has 61.5 mg methanol in the aspartame molecule, so 100 ml
diet soda has 6.15 mg methanol, so the brandy has 24.4 times more methanol
than diet soda. A pound of fruit gives about as much methanol as 2 L
(nearly 6 cans) diet soda. ]

I urge Channing Laboratory and its participating universities to rapidly
mount an in-house study to study the Nurses Health Study database for the
hundreds of nurses who are long-term users, above 6 cans diet drinks daily,
for correlations with every disease, as well as ubiquitous co-factors like
wine and liquor, cigarette smoke, and fruits and vegetables. It could
vastly serve the world public health to make the initial findings widely
available immediately. The disparaged issue of aspartame toxicity could be
swiftly made legitimate, and the resulting progress on all levels remarkably
accelerated.

A single scientist could do this.

Comments pro and con are welcome. A convenient venue would be
the moderated Usenet group: bionet.toxicology.


http://groups.yahoo.com/group/aspartameNM/message/1184
corporate corruption of health sciences, International Journal of
Occupational and Environmental Health, entire issue, 2005 Oct-Dec: Gary N
Greenburg, OEM-L: aspartame (methanol, formaldehyde, formic acid) toxicity,
Murray 2005.07.14

http://groups.yahoo.com/group/aspartameNM/message/1155
continuing aspartame debate in British Medical Journal, John Biffra, Bob
Dowling, Nick Finer, Ian J Gordon: Murray 2005.02.09 rmforall

http://groups.yahoo.com/group/aspartameNM/message/782
RTM: Smith, Terpening, Schmidt, Gums:
full text: aspartame, MSG, fibromyalgia 2002.01.17 rmforall
Jerry D Smith, Chris M Terpening, Siegfried OF Schmidt, and John G Gums
Relief of Fibromyalgia Symptoms Following
Discontinuation of Dietary Excitotoxins.
The Annals of Pharmacotherapy 2001; 35(6): 702-706.
Malcolm Randall Veterans Affairs Medical Center, Gainesville, FL, USA.
BACKGROUND: Fibromyalgia is a common rheumatologic disorder that is
often difficult to treat effectively.
CASE SUMMARY: Four patients diagnosed with fibromyalgia syndrome
for two to 17 years are described.
All had undergone multiple treatment
modalities with limited success. All had complete, or nearly complete,
resolution of their symptoms within months after eliminating monosodium
glutamate (MSG) or MSG plus aspartame from their diet.
All patients were women with multiple comorbidities
prior to elimination of MSG.
All have had recurrence of symptoms whenever MSG is ingested.

Siegfried O. Schmidt, MD Asst. Clinical Prof.
Community Health and Family Medicine, U. Florida, Gainesville, FL
Shands Hospital West Oak Clinic Gainesville, FL 32608-3629
352-376-5071
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