http://groups.yahoo.com/group/aspartameNM/message/804

RTM: Hetle & Eltervaag: 2001 thesis
abstract: aspartame brain damage in mice:
Sonnewald 1995 study full text 2.17.2 rmforall

For thesis in Norwegian, mailed by regular mail, contact:
Anne Værnes <anne.varnes@...>

"Cola light, one calorie" men hva med jhernen?
Hovedfagoppgave hosten 2001
Utfort av Arnstein Eltervaag og Elisabeth Hetle
Det medisinske fakultet Institutt for kliniske nevrofag
Trondheim Norway 10.desember 2001

The 48-page thesis has 35 references, and includes an English abstract.
Faculty and helpers listed in the Forword are:
Ursula Sonnewald (with 134 items in PubMed since 1988, showing a
distinguished research career in biochemical studies of neurotoxins--
one of her studies on aspartame, published 1995 with three partners,
Tomm Muller, Geirmund Unsgard, and S.B. Peterson, is given in full at
the end of this post, with 18 references, and obviously presents much
the same laboratory technique as applied in 2001 in the thesis.),
Hong Qu [female qu.hong@... ], and
Bente Urfjell. Obviously, this team has the experience, facilities,
funding, faculty support, and motivation to study the biochemistry of
aspartame toxicity in detail.

ABSTRACT

Introduction: Aspartame (ASM) is a product that was originally made for
diabetics, but today ASM is widely used by healthy people
as an artificial sweetener in many food products.

Purpose: The main goal with this research was to see whether ASM was
harmful to brain cells (cerebellar granule cells).
We wanted to check if the damage to the neurons is connected to
the N-methyl-D-aspartate (NMDA)-receptors on these cells.

Procedure: Brain cells from 7 day old mice were used. They were
cultured in 24 Petri well dishes, and different quantities of ASM were
added.
After 7 days, the cultures were analysed by two different tests:
Lactate dehydrogenases (LDH) test, which gives a picture of cell death
(LDH leakage to the medium in which the cells were cultured).
3-[4,5-dimethylthiazol-2yl]-2,5-diphenyltetrazolium bromid (MTT) test,
which can be used to analyse mitochondrial activity in living cells.
To test whether the NMDA-receptor was involved in the damage done by
ASM, the receptor was blocked by
(±)-2-amino-5 phosphonopentanocid (AP5).

Results: Our results showed damage/cell death from an added quantity
of 0.06 mg/ml ASM each day for 4 days.
As a comparison there is 0.24 mg/ml ASM in Cola Light.
MTT- and LDH-tests showed damage to the neurons at an added
quantity of 1.5 and 3.00 mg/ml ASM after 22 hours of incubation.
The results also show that ASM is in part acting through the
NMDA-receptor because AP5 reduced or
blocked the damage to the granule cells.

Conclusion: In light of these results, our conclusion is that in order
to be on the safe side, it should be warned against use of ASM
as a food additive, maybe especially in products consumed by children,
because NMDA-receptors and the synapses involved
also are connected to learning.
**********************************************************

http://www.dagbladet.no/print/?/dinside/2001/12/17/301529.html
[A major newspaper in Norway]

[Photo caption]
FARLIG FOR HJERNEN?
Medisinstudent Elisabeth Hetle (32) har sluttet å drikke lettbrus, mens
medstudent Arnstein Eltervaag (40) aldri har drukket lettbrus.

I edited this into a fairly accurate English version:

[Caption for photo]
DANGER FOR BRAIN?
Medical student Elisabeth Hetle (32) has stopped using aspartame diet
sodas, while fellow student Arnstein Eltervaag (40) has never used them.

You can also read this article at: [article on newspage]
http://www.dagbladet.no/dinside/2001/12/17/301529.html
Dagbladet © 2001:

hetle@... eltervaa@...
**********************************************************

NTNU: Norges teknisk-naturvitenskapelige universitet:
[English] Norwegian University of Science and Technology
NO-7491 Trondheim, Norway
Phone.: +47 73595000, Fax: +47 73595310

http://www.ntnu.no/indexe.php
Creative, Constructive, Critical:
These are the keywords in our
strategy. As the name states the
Norwegian University of Science and
Technology, NTNU, is a centre for
technological education and research
in Norway, with a solid foundation in
the natural sciences.
This tradition is interwoven with
broadly based expertise in the
classical university disciplines of the
humanities, medicine and the social sciences.
At the same time, NTNU offers the
widest range of education in art
subjects; music, the visual arts and
architecture, of all the universities in Norway.
At NTNU we strive to encourage soaring imagination and restless
curiosity. Our ambition is to promote a creative interplay between all
forms of human intellectual activities, the arts,
the natural and social sciences, and technology.
Our interdisciplinary efforts are inspired by the consummate
Renaissance man Leonardo da Vinci,
who unified areas of study previously seen as distinct.
NTNU is an institution that provides stimulating challenges for those
who want to explore new approaches. NTNU is about leading the field.

Faculty of Medicine: http://www.medisin.ntnu.no/eng/
The faculty does research in all the medical disciplines: basal,
paraclinical and clinical subjects. Medical technology is an important
area of cooperation for the faculty.
Academic posts: 146. Doctoral students: 77.

Besøksadresse: Olav Kyrres g. 3, Medisinsk teknisk forskningssenter
Postadresse: Medisinsk teknisk forskningssenter, N-7489 Trondheim
Tlf.: 73 59 88 59, faks: 73 59 88 65, E-post: dmf-post@...
**********************************************************

http://www.sintef.no/units/unimed/mr/Staff/ursula.htm
URSULA SONNEWALD Ph.D., Professor
Dept. of Pharmacology and Toxicology,
Olav Kyrres g.3,
Norwegian University of Science and Technology,
N-7489 Trondheim Norway
phone: (+47) 73 59 04 92, fax: (+47) 73 59 86 55, e-mail:
Ursula.sonnewald@...

http://www.ntnu.no/forskning/publikasjoner98/dmf_farmakologi
NTNU - det medisinske fakultet
Institutt for farmakologi og toksikologi
**********************************************************

http://www.oslo.sintef.no/annual/94/eng/14.html
(photo) First-aid for brain cells
Research at SINTEF UNIMED´s MR Centre has given us a better
understanding of what takes place in brain cells when their oxygen
supply is reduced or cut off. As a result of its work, the Centre has
been awarded a research contract by a leading Japanese drug company.
In SINTEF UNIMED´s laboratory, Ursula Sonnewald has cultivated mouse
brain cells.

The MR Centre is now testing new drugs which are being developed by
Yamanouchi Pharmaceuticals. The manufacturer hopes that these medicines
will enable doctors to reduce brain damage in stroke patients and others

who are suffering from reduced or blocked oxygen supply to the central
nervous system (CNS).
SINTEF UNIMED will test the new drugs by means of in vitro studies and
animal tests.

A lack of oxygen in the brain can lead to lasting damage since brain
cells die, and because the bodily functions that are controlled by these

cells are put out of operation. Damage of this sort can occur in
patients who have suffered strokes, heart failure, in newborn children
with paranatal injuries and people who have been rescued from drowning.

However, cells whose oxygen supply is cut for a short period are capable

of surviving for a day after they are damaged. In theory, this means
that it ought to be possible to save many cells before the damage
becomes permanent, if only we knew just what happens in the cells at
this time.

The photomicrograph below shows a collection of nerve cells that have
built up a neural network in the petri dish. In the background we can
see glial support cells (astrocytes).

Biochemical changes

In collaboration with Professor Geirmund Unsgård of the University of
Trondheim, Dr. Tomm Müller of Trondheim Regional Hospital
and scientists from the Pharmaceutical College of Denmark,
Dr. Ursula Sonnewald at the MR Centre
has been studying cell cultures from mice and rats. Their
studies have provided new understanding of the biochemical changes that
take place during the first few hours after the oxygen supply to the
brain has been cut off.

Dr. Sonnewald and her partners have demonstrated differences in
mechanisms of injury in the nerve cells themselves (the neurones) and
the surrounding glial cells (the astrocytes) that keep the neurones
alive. For this purpose she has used a spectroscopic analysis technique
that utilizes nuclear magnetic resonance (NMR). In SINTEF UNIMED´s cell
experiments for Yamanouchi the effects of the drugs on injured brain
cells in cell cultures are studied by means of the same technique.

Brain images from living animals

A parallel study at SINTEF UNIMED is looking at the effects of drugs on
the brains of living rats with the aid of NMR-based imaging.
The special method that is being used in this part of the study was
developed by Dr. Müller in the course of his doctoral studies, in
collaboration with Dr.
Olav Haraldseth and Dr. Richard Jones, both at SINTEF UNIMED. This
method makes it possible to identify those regions of the CNS that
undergo alterations when the blood supply to these animals' brains
stops. The images also show the size of the areas involved.

When the animals are given medication, the images can show the extent
to which the injuries disappear. In this way the technique can tell us
whether a given drug is capable of crossing the blood-brain barrier, as
it must do if it is to have its intended effect on the central nervous
system.

Contact persons: Ursula Sonnewald Olav Haraldseth
**********************************************************
[ 13C and 45Ca are radioactive isotopes of carbon and calcium, used to
trace biochemical reactions.]

1
Bakken, Ingen Johanne; White, Linda R.; Aasly, Jan; Unsgård, Geirmund;
Sonnewald, Ursula;
<U-13C> aspartate metabolism in cultured cortical astrocytes and
cerebellar granule neurons studied by NMR spectroscopy.
GLIA. Wiley-Liss, Inc. 23, 271-277 1998

2
Bakken, Inger Johanne; White, Linda R.; Unsgård, Geirmund; Aasly, Jan;
Sonnewald, Ursula
<U-13C>-glutamate Metabolism in Astrocytes During Hypoglycemia and
Hypoxia.
Journal of Neuroscience Research. Wiley-Liss, Inc. 51, 636-645 1998

3
Håberg, Asta; Qu, Hong; Bakken, Inger Johanne; Sande, Leif Magne; White,

Linda R.; Haraldseth, Olav; Unsgård, Geirmund; Aasly, Jan;
Sonnewald, Ursula
In vitro and ex vivo 13C-NMR spectroscopy studies of pyruvate
recycling in brain.
Developmental Neuroscience. S. Karger AG 20, 389-398 Basel 1998

4
Håberg, Asta; Qu, Hong; Haraldseth, Olav; Unsgård, Geirmund; Sonnewald,
Ursula
In vivo Injection of 1-13C Glucose and 1,2-13C Acetate Combined With Ex
Vivo 13C Nuclear Magnetic Resonance Spectroscopy:
A Novel Approach to the Study of
Middle Cerebral Artery Occlusion in the Rat.
Journal of Cerebral Blood Flow and Metabolism. Lippincott Williams &
Wilkins 18: 11, 1223-1232 Philadelphia 1998

5
McKenna, Mary C.; Sonnewald, Ursula; Huang, Xueli; Stevenson, Joseph;
Johnsen, Svein F.; Sande, Leif M.; Zielke, H. Ronald
-a-Ketoisocaproate alters the production of both lactate and
aspartatefrom <U-13C>glutamate in astrocytes : a 13C NMR study.
Journal of Neurochemistry. Lippincott-Raven Publishers 70, 1001-1008
Philadelphia 1998

9
Sonnewald, Ursula; Sonnewald, Ursula; Akiho, Hiraku; Koshiya, Kazuo;
Iwai, Akihiko
Effect of orotic acid on the metabolism of cerebral cortical astrocytes
during hypoxia and reoxygenation : an NMR spectroscopy study.
Journal of Neuroscience Research. Wiley-Liss, Inc. 51: 1, 103-108 1998

14
Waagepetersen, H.S.; Bakken, I.J.; Larsson, O.M.; Sonnewald, Ursula;
Schousboe, A.
Comparison of lactate and glucose metabolism in cultures neocortical
neurons and astrocytes using 13C-NMR spectroscopy.
Developmental Neuroscience. S. Karger AG 20, 310-320 Basel 1998

15
Waagepetersen, Helle S.; Bakken, Inger J.; Larsson, Orla M.; Sonnewald,
Ursula; Schousboe, Arne
Metabolism of Lactate in Cultured GABAergic Neurons Studied by 13C
Nuclear Magnetic Resonance Spectroscopy.
Journal of Cerebral Blood Flow and Metabolism.
Lippincott-Raven Publishers 18: 1, 109-117 Philadelphia 1998

31
Qu, Hong; Håberg, Asta; Sæter, Oddbjørn; Haraldseth, Olav; Unsgård,
Geirmund; Sonnewald, Ursula
Pyruvate recycling.
Journal of Neurochemistry. Lippincott-Raven 71 USA, 1998

32
Schousboe, Arne; Gegelashvili, Georgi; Sonnewald, Ursula
Role of astrocytes in glutamate metabolism during neurotransmission.
Journal of Neurochemistry. Raven Lippincott 71 USA, 1998

33
Sonnewald, Ursula; Håberg, Asta; Qu, Hong; Sæter, Oddbjørn; Haraldseth,
Olav; Unsgård, Geirmund
Stroke, the metabolic approach.
Journal of Neurochemistry vol. 71. Lippincott-Raven USA, 1998
***********************************************************

http://www.phys.ntnu.no/instdef/grupper/biosystemer/qu.hong/
qu.hong@...
http://www.ntnu.no/doktorgrader/dr.scient/02.02/qu.htm
(photo of woman) Cand.scient. Hong Qu (30) fra Shenyang, Kina, har
studert vekselvirkninger mellom celler i hjernen (astrocytter og
nevroner) i sin doktoravhandling ved Norges teknisk-naturvitenskapelige
universitet, NTNU.
***********************************************************

NEUROPHARMACOLOGY AND NERUOTOXICOLOGY Volume 6 1995 (PP318-320) Rapid
Communications of Oxford Ltd

Effects of aspartame on Ca influx and LDH leakage from nerve cells in
culture
Ursula Sonnewald, Tomm Muller, Geirmund Unsgard, S.B. Peterson
MR-Centre, SINTEF UNIMED, N-7034
Trondheim; University of Trondheim, Dept. of Neurosurgery,
University Hospital N-7006
Trondheim; Norwegian Institute of Tecnology, Drpt. of Biotecnology,
N-7034 Trondheim, Norway

Aspartame (ASM), an artificial sweetener, was shown to dose dependently
increase CA influx into and lactate dehydrogenase (LDH) leakage from
murine brain cell cultures.
Astrocytes were more resistant than neurones to the effects of ASM.
In cerebellar granule neurones, a 20% increase in calcium
was found after an incubation time of 22 h in the presence of 0.1 mM
ASM;
at 0.5 mM concentration, calcium influx increased 40% compared with
control cultures.
At a concentration of 10mM, influx was increased 13-fold after 5 h.
Morphological appearance as judged by phase contrast microscopy was
first visibly affected after exposure to 1mM ASM for 22 h.
Citrate, another food additive, was included in the study to demonstrate
that cerebellar granule neurones could tolerate 10mM additions to the
medium and citrate did not cause Ca influx or morphological changes in
neurones after 22 h.
LDH leakage, a sign of severe cell damage, was observed at 1 mM
concentrations of ASM after 22 h.
Cerebral astrocytes on the other hand were more resistant and showed
morphological changes, increased calcium influx and LDH leakage
first at 5 mM concentrations of ASM.

Key words: Aspartame, Neurotoxicity; Cerebellar granule neurones;
Lactate dehydrogenase leakage; Calcium influx

INTRODUCTION
Aspartame (L-aspartyl--L-phenylalanine methyl ester, ASM) is a widely
used artificial sweetener in soft drinks and low calorie food.
There have been reports of adverse neurological effects
such as headache (1),
insomnia and seizures after ingestion of aspartame, which may be
attributed to the alterations in regional concentrations of
catecholamines.(2)
Brain phenylalanine and tyrosine were increased
following ASM ingestion. (3)
Studies using radioactively labelled aspartame in comparison with
labelled methanol, aspartame and phenylalanine have shown the 30-40% of
the total dose of aspartame of the labelled components remains in the
body after 8 h; the remainder is primarily s
ecreted through expired air. (4)
Analysis of tissue distribution of orally administered isotopically
labelled aspartame in the rat showed part of the label remaining in the
brain for up to 24 h. (5)
From these studies it was not possible to
determine whether ASM or its degradation products reached the brain.

Both aspartate (6) and aspartame (7) have been shown to have excitatory
activity. Olney et al (8) have shown that systemic administration of
glutamatae, an excitatory amino acid, produced brain damage in a number
of animal species including primates, and excitotoxic analogues such as
aspartame had the same effects. (9)

In order to investigate potential toxicity of aspartame on brain cells,
lactate dehydrogenase leakage and (45) Ca influx into astrocytes and
neurones were measured after incubation with
varying concentrations of aspartame.

Materials and Methods

Plastic tissue culture dishes were purchased form NUNC A/S (Denmark),
fetal calf serum from Seralab (Sussex, UK), poly-L-lysine (mol wt.> 300
000) and amino acids from Sigma (St. Louis, MO) ; 45Ca was from
Amersham. All other chemicals were of the purest grade available from
regular commercial sources.

Cortical astrocytes were cultured essentially as described by Hertz et
al. (10) Prefrontal cortex was taken from newborn NMRI mice and passed
through Nitex nylon sieves (80 um pore size) into a slightly modified
Dulbecco's medium (DMEM) containing 20% (v/v) fetal calf serum and
plated in NUNC 3 cm culture dishes. Medium was changed twice a week.
Cells were used for experiments after 2-3 weeks in culture.
Cerebellar granule cells were prepared from 7-day-old mice; (11) they
have been shown to possess NMDA receptors (12) and are useful in the
study of neurotoxicity. (12)
Tissue samples of cerebella were exposed to mild trypsinization
followed by trituration in a DNAse solution containing a soyabean
trypsin inhibitor. Cells were suspended (2-3 x 106 cells ml-1)
in a slightly modified DMEM with 10% (v/v fetal calf serum.
Cytosine arabinoside (20 uM) was added after 48 h to prevent astrocyte
proliferation.
Cells were used after 7 days in culture. Prior to experiments, the
incubation medium was removed and substituted with Hanks balanced salt
solution without MG2+ (HBBS) containing 1.5 uCi ml-1 (45)Ca.
The experiments were terminated by the removal of the incubation
medium. The cells were washed five times with ice-cold
phosphate-buffered saline containing 25 mM MgCl2 to displace (45) Ca
bound extra-cellularly.
The cells were lysed in 0.5 M HCL and the (45) Ca content
was determined by liquid scintillation spectrometry.
When appropriate, cell integrity in the cultures was assessed by
determination of leakage of lactate dehydrogenase (LDH< EC 1.1.27) from
cells into the medium, using a diagnostic kit supplied by Sigma Chemical
(catalogue no. DG 1340-K).
LDH was measured in cell extracts and medium and expressed
as percentage of total LDH ((14)

Results and Discussion

Aspartame has been shown to dose-dependently inhibit L-(3H) glutamate
binding to the N-methyl-D-aspartame (NMDA) receptor in a synaptosomal
preparation from rat brain. (7)
The NMDA receptor is an ionotropic
glutamate receptor mediating calcium influx into neurones.
Aspartate, a constituent of ASM, is a potent NMDA agonist and has been
shown to induce widespread late neuronal degeneration. (14)
Delayed cell death mediated by the NMDA receptor depended on the
presence of extracellular calcuium. (15-17)
Thus the present study was undertaken to evaluate the effect of ASM
on primary nerve cell cultures in terms of calcium influx.
Furthermore measurement of LDH activity released to the extracellular
media has been found to be a quantitative method for determining
neuronal cell injury. (18)
Table 1 shows that ASM dose-and time-dependently increase calcium
influx into and LDH leakage from cerebellar granule neurones.
No effect was detected at 0.1 mM, but at 0.5 mM ASM LDH leakage was
increased slightly and at a concentration of 5 mM LDH leakage was
increased by a factor of 2.5 after 22 h (Table 1).
After this time cells had detached from the culture dishes and
intracellular (45)Ca could not be determined.
At 10 mM, calcium influx was increased 13-fold after a 5 h
incubation (Table 2).
Citrate, another food additive, was included in the
study to demonstrate that cerebellar granule neurones could tolerate
addition of organic substances at 10 mM concentration to the medium and
citrate did not cause (45) Ca influx or morphological changes in
neurones;
however, deleterious effects on astrocytes were seen.
The above findings further confirm the hypothesis of Pan-How et al (7)
that the neurotoxicity produced by ASM is mediated by a calcium coupled
receptor.
In the case of cerebellar granule neurones it is likely to be an NMDA
receptor-mediated effect.
The excitotoxin responsible for this effect could either be free
aspartate (an NMDA receptor agonist) derived from proteolytic cleavage
of ASM or ASM directly.
Astrocytes on the other hand are not believed to have NMDA receptors
and the observed calcium influx at 5 mM ASM (Table 1) must therefore be
mediated through a different mechanism.
LDH leakage, a sign of cell damage, was also observed in astrocytes
(Table 1). Thus it has been shown that ASM has adverse effects both on
glia and neurones in culture.

Clearly the concentrations used in these studies are not likely to be
physiological, but subpopulations of neurones might be affected by
lower doses, and long term exposure to low concentrations might
produce cumulative irreversible damage.
Based on the results presented here, we cannot draw any conclusions for
the in vivo situation, there is the need for additional in vitro and in
vivo studies, to evaluate the safety of this food additive that is
consumed in increasing amounts by adults and children.

References
1. Johns Dr. Migraine provoked by aspartame. N Engl J Med 315, 456
(1986)

2. Coulomb, RA and Sharma RS. Neurobiochemical alterations induced by
the artificial sweetener aspartame. Toxicol Parmacol 83d, 79-85 (1986)

3. Fernstrom JD, Fernstrom MH and Gillis MA.
Acute effects of aspartame on large neutral amino acid and monoamines
in rat brain. Life Sci 32, 1651-1658 (1983)

4. Opperman JA. Aspartame metabolism in animals. In Stegink LD and
Filer Jr. eds. Aspartame Physiology and Biochemnistry.
New York: Marcel Dekker, 1984: 161-200.

5. Matsuzawa Y and O'Hara Y. Tissue distribution of orally administered
isotopically labelled aspartame in the rat. In. Stegink LD and Filer
Jr. eds. Aspartame Physiology and Biochemistry.
New York: Marcel Dekker, 1984; 161-200

6. Watkins JC. Excitatory amino acid and central synaptic transmision .
Trends Pharmacol 5 373-376 (1984)

7. Pan-Hou H, Ohe Y, Sumi M et al. Effect of aspartame on NMDA sensitive

L-(3H)glutamate binding sites in rat brain synaptic membranes.
Brain Res 520, 351-353 (1990)

8. Olney Jw. Sharpe LG and Feigin Rd.
Glutamate-induced brain damage in infant primates.
J Neuropathol Exp eurol 31, 464-488 (1972)

9. Olney JW, Sharpe LG and Feigin RD. Glutamate-induced brain damage in
infant primates. J Neuropathol Exp Neurol 31, 464-488 (1972)

10. Hertz l, Juurlink BHG, Hertz E et al. Preparation of primary
cultures of mouse (rat) astrocytes. IN: Shahar A, De Vellis J,
Vernadakis A, Haber B, eds.
A dissection and Tissue Culture Manual of the Nervous System
New York: Liss, 1989:105-108

11. Schousboe A, Meier E, Drejer J et al. Preparation of primary
cultures of mouse (rat) cerebellar granule cells. In Shahar A, De
Vellis J, Vernadakis A. Haber B, eds.
A Dissection and Tissue Culture Manual of the Nervous System.
New York: Liss, 1989: 183-186

12. Lysko PG, Cox JA, Vignano MA et al. Excitatory amino acid
neurotoxicity at the N-methyl-E-aspartame receptor
in cultured neurones; pharmacological characterization,
Brain Res 499, 258-266 (1989)

13. Frandsen AA and Schousbor A. Time and concentration dependency of
the toxicity of excitatory amino acids on cerebral neurones in primary
culture. Neurochem Int 10, 583-591 (1987)

14. Choi DW. Non-NMDA receptor-mediated neuronal injury
in Alzheimer's disease? Neurobial Aging 10, 605-606 (1989)

15. Hartly DM, Kurth MC , Bjerkness L et al.
Glutamate receptor-induced (45) Ca2+ accumulation in cortical cell
culture correlates with subsequentneuronal accumulation in cortical cell
culture correlates with subsequent neuronal degeneration.
J Neursci 13 1993-2000 (1993)

16. Sijesjo BK and Bengtsson F. Calcium fluxes, calcium antagonists,
and calcium-related pathology in brain ischemia, hypoglycemia, and
spreading depression: A unifying hypothesis.
J Cereb Blood Flow Metab 9, 127-140 (1989)

17.Eimerl S and Schramm. The quantity of calcium that appears to induce
neuronal death. J Neurochem 62 1223-1226 (1994)

18. Koh JY and Choi DW. Quantitative determination of glutamate
mediated cortical neuronal injury in cell culture by lactate
dehydrogenase efflux assay. J Neurosci Methods 20, 83-90 (1987)

Acknowledgements: Thisresearch was supported by the Research Council
of Norway. The use of the animal facilities at the University Hospital

in Trondheim are gratefully acknowledged.
Received 26 October l994; accepted 25 Nov l994

Table 1. The effect of aspartame on calcium influx and LDH leakage in
cerebellar granule neurones and cortical astrocytes
**********************************************************

Rich Murray, MA Room For All rmforall@...
1943 Otowi Road, Santa Fe NM USA 87505 505-986-9103

http://groups.yahoo.com/group/aspartameNM/messages for 804 posts
http://groups.yahoo.com/group/aspartameNM/message/657 45K post
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http://www.dorway.com/tldaddic.html 5-page review
"Aspartame (NutraSweet) Addiction"
H.J. Roberts in "Townsend Letter", Jan 2000 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 $ 85.00 postpaid data from 1200 cases
available at http://www.amazon.com
over 600 references from standard medical research
http://www.aspartameispoison.com/contents.html 34 chapters
http://groups.yahoo.com/group/aspartameNM/message/790
RTM: Moseley:
review Roberts "Aspartame Disease: An Ignored Epidemic" 2.7.2 rmforall

http://groups.yahoo.com/group/aspartameNM/message/652
Ann Pharmacother 2001 Jun;35(6):702-6
Relief of fibromyalgia symptoms following
discontinuation of dietary excitotoxins.
terpening@... cterpeni@...
Smith JD, Terpening CM, Schmidt SO, Gums JG.
Malcolm Randall Veterans Affairs Medical Center, Gainesville, FL, USA.
gums@... siggy@...

http://groups.yahoo.com/group/aspartameNM/message/782
RTM: Smith, Terpening, Schmidt, Gums:
full text: aspartame, MSG, fibromyalgia 1.17.2 rmforall

http://groups.yahoo.com/group/aspartameNM/message/804
RTM: Hetle & Eltervaag: 2001 thesis
abstract: aspartame brain damage in mice:
Sonnewald 1995 study full text 2.17.2 rmforall
**********************************************************