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


Pharmacol Res. 2007 May 13; [Epub ahead of print]
The effect of aspartame on acetylcholinesterase activity in hippocampal
homogenates of suckling rats.
Simintzi I,
Schulpis KH,
Angelogianni P,
Liapi C,
Tsakiris S.
Department of Experimental Physiology,
Medical School, University of Athens,
P.O. Box 65257, GR-15401 Athens, Greece.

BACKGROUND:
Neurological disturbances have been implicated with aspartame (ASP) consumption,
and the cholinergic system with acetylcholinesterase (AChE) seems actively
involved.

AIM:
To evaluate the effect of ASP and its metabolites on rat hippocampal AChE
activity.

METHODS:
Hippocampal homogenate or pure enzyme AChE (eel E. electricus) was incubated
with the sum or each of ASP components, phenylalanine (Phe), aspartic acid (asp)
and methanol (MeOH) for 1h at 37 degrees C.

AChE activity was measured spectrophotometrically.

RESULTS:
Incubation of rat tissue or pure enzyme with the sum of ASP metabolites in
concentrations in CSF
(the concentrations were calculated according to the CSF/plasma concentration
ratios)
following 150 or 200mgkg(-1) of ASP consumption,
resulted in significant enzyme activity reductions of 25 and 31%
for hippocampal AChE and 11% (p<0.01) and 19% for pure enzyme, respectively.

Aspartic acid concentrations of 0.42 or 0.56mM
significantly reduced the enzyme activities by 13 and 20%
for hippocampal AChE and 15 and 18% for pure enzyme, respectively.

Phe concentrations of 0.042 or 0.083mM decreased the enzyme activity
by 12% (p<0.01) and 20% (p<0.001) for hippocampal AChE
and 15 and 18% (p<0.001) for pure enzyme, respectively.

Methanol concentrations of 0.60 or 0.80mM remarkably inhibited
hippocampal AChE by about 18 and 22%
and pure enzyme by about 14 and 20%, respectively.

CONCLUSIONS:
Low concentrations of ASP components had no effect on hippocampal and pure AChE
activity,
whereas high or toxic concentrations remarkably decreased both enzyme
activities.

Muscarinic symptoms may be related to the latter concentrations of ASP
metabolites. PMID: 17580119

Kleopatra H. Schulpis, MD, PhD. Institute of Child Health,
Aghia Sophia Children's Hospital, GR-11527 Athens (Greece)
Tel. +30 1 7708291, Fax +30 1 7700111 inchildh@...


Metab Brain Dis. 2006 Mar; 21(1): 21-8. Epub 2006 May 6.
The effect of in vitro homocystinuria on the suckling rat hippocampal
acetylcholinesterase.
Schulpis KH,
Kalimeris K,
Bakogiannis C,
Tsakiris T,
Tsakiris S.
Inborn Errors of Metabolism Department,
Institute of Child Health Research Center,
Aghia Sophia Children's Hospital, Athens, Greece.

Homocystinuria is due to enzymatic deficiencies
resulting in elevated blood levels of homocysteine (Hcy),
homocystine (Hci),
and/or methionine (Met)
and the clinical presentation of mental retardation, seizures, and
cardiovascular disease.

Since these symptoms may be closely implicated with acetylcholinesterase (AChE)
activity,
we aimed to investigate whether this metabolic disorder affects the hippocampal
AChE activity
in 21 days suckling Wistar rat hippocampus.

Various concentrations of Hcy, Hci (0.05-0.5 mM), or Met (0.05-2 mM)

as well as Mixture A (Mix A) (0.3 mM (Hcy)+0.2 mM (Hci)+1.0 mM (Met) = in vitro
cystathionine beta-synthase deficiency homocystinuria),

Mix B1 (Hcy 0.3 mM + Hci 0.2 mM = in vitro severe methylenetetrahydrofolate
reductase deficiency homocystinuria)

or Mix B2 (Hcy 0.1 mM+Hci 0.05 mM = in vitro mild methylenetetrahydrofolate
reductase deficiency homocystinuria)

were preincubated with homogenized hippocampii or with eel Electrophorus
electricus pure AChE.

AChE was evaluated spectrophotometrically.

Hcy or Met stimulated hippocampal AChE by 50% (p < 0.001)
at low concentrations of the amino acids (up to 0.3-0.5 mM),
whereas Hci inhibited the enzyme by 40% (p < 0.001).

Mix A, Mix B1, or Mix B2 activated hippocampal AChE
by 40, 30, (p < 0.001), and 12% (p < 0.01), respectively.

In contrast, the S-containing amino acids, Mix A, Mix B1, Mix B2
failed to affect the pure AChE activity.

CONCLUSIONS:
a) The presence of -SH group in Hcy and Met may result in hippocampal AChE
stimulation
and the redox isomer Hci in the inhibition of the enzyme,
probably by producing free radicals,

and b) The SH-amino acids seem to affect the hippocampal enzyme indirectly,
possibly by lipid(s)-protein modifications(s)
and Hci by inducing oxidative stress,
since no effect was observed on pure AChE activity.
PMID: 16773467


http://groups.yahoo.com/group/aspartameNM/message/1279
all three aspartame metabolites harm human erythrocyte [red blood cell]
membrane enzyme activity, KH Schulpis et al, two studies in 2005,
Athens, Greece, 2005.12.14: 2004 research review, RL Blaylock:
Murray 2006.01.14

"High or abuse concentrations of ASP hydrolysis products significantly
decreased the membrane enzyme activity,
which was completely or partially prevented by L-cysteine
or reduced GSH."

[ Definition of Erythrocyte
Erythrocyte:
A cell that contains hemoglobin and can carry oxygen to the body.
Also called a red blood cell (RBC).
The reddish color is due to the hemoglobin.
Erythrocytes are biconcave in shape,
which increases the cell's surface area
and facilitates the diffusion of oxygen and carbon dioxide.
This shape is maintained by a cytoskeleton
composed of several proteins.
Erythrocytes are very flexible
and change shape when flowing through capillaries.
Immature erythrocytes, called reticulocytes,
normally account for 1-2 percent of red cells in the blood. ]


Eur J Clin Nutr. 2005 Dec 14; [Epub ahead of print]
The effect of L-cysteine and glutathione on inhibition of
Na(+), K(+)-ATPase activity by aspartame metabolites
in human erythrocyte [red blood cell] membrane.
Schulpis KH, Kleopatra H. Schulpis, MD, PhD.
Institute of Child Health, Aghia Sophia Children's Hospital,
GR-11527 Athens (Greece) +30 1 7708291, Fax +30 1 7700111 inchildh@...,
Papassotiriou I, biochem@...,
Parthimos T,
Tsakiris T,
Tsakiris S. Stylianos Tsakiris. stsakir@...,
1 Institute of Child Health, Research Center,
'Aghia Sophia' Children's Hospital, Athens, Greece.
ggbriass@..., ersi_voskaridou@...,
mmoschov@..., siahanidou@...,


Background:
Reports have implicated Aspartame
(N-L-a-aspartyl-L-phenylalanine methyl ester, ASP)
in neurological problems.

Aim:
To evaluate Na(+), K(+)-ATPase activities in human erythrocyte
[red blood cell] membranes
after incubation with the ASP metabolites,
phenylalanine (Phe),
methanol (MeOH) and
aspartic acid (Asp).

Methods:
Erythrocyte [red blood cell] membranes
were obtained from 12 healthy individuals and
were incubated at 37 degrees C for 1 h
with the sum or each of the ASP metabolites separately,
which are commonly measured in blood after ASP ingestion.

Na(+), K(+)-ATPase and Mg(2+)-ATPase activities were measured
spectrophotometrically.

Results:
Membrane Mg(2+)-ATPase activity was not altered.

The sum of ASP metabolite concentrations corresponding
to 34, 150 or 200 mg/kg of the sweetener ingestion
resulted in an inhibition of the membrane
Na(+), K(+)-ATPase by -30, -40, -48%, respectively.

MeOH concentrations of 0.14, 0.60 or 0.80 mM
decreased the enzyme activity by -25, -38, -43%, respectively.

Asp concentrations of 2.80, 7.60 or 10.0 mM
inhibited membrane Na(+), K(+)-ATPase by -26, -40, -46%,
respectively.

Phe concentrations of 0.14, 0.35 or 0.50 mM
reduced the enzyme activity by -24, -44, -48%, respectively.

Preincubation with L-cysteine or reduced glutathione (GSH)
completely or partially restored
the inhibited membrane Na(+), K(+)-ATPase activity
by high or toxic ASP metabolite concentrations.

Conclusions:
Low concentrations of ASP metabolites had no effect
on Na(+), K(+)-ATPase activity.

High or abuse concentrations of ASP hydrolysis products significantly
decreased the membrane enzyme activity,
which was completely or partially prevented by L-cysteine
or reduced GSH. [reduced glutathione]

European Journal of Clinical Nutrition advance online publication,
14 December 2005; doi:10.1038/sj.ejcn.1602355. PMID: 16391576



http://groups.yahoo.com/group/aspartameNM/message/1213
aspartame (methanol, phenylalanine, aspartic acid) effects, detailed
expert studies in 2005 Aug and 1998 July, Tsakiris S, Schulpis KH,
Karikas GA, Kokotos G, Reclos RJ, et al, Aghia Sophia Children's
Hospital, Athens, Greece: Murray 2005.09.09

[ The lowest dose level tested, 34 mg aspartame per kg body weight,
well below the FDA daily human limit of 50 mg/kg, 16 12-oz cans,
caused enzyme activity reduction by -33% in human red blood cell
membranes. ]

However, a missed opportunity in both studies is that the inevitable,
extremely and cumulatively toxic products of methanol in the human
body, formaldehyde and formic acid, which are responsible for the
toxicity of methanol, were not independently tested.

" It is concluded that low concentrations of ASP metabolites had no
effect on the [human red blood cell] membrane enzyme activity,
whereas high or toxic concentrations partially or remarkably decreased
the [human red blood cell] membrane AChE activity, respectively.
Additionally, neurological symptoms, including learning and memory
processes, may be related to the high or toxic concentrations of the
sweetener metabolites. " ]

Pharmacol Res. 2005 Aug 26; [Epub ahead of print]
The effect of aspartame metabolites on human [red blood cell]
erythrocyte membrane acetylcholinesterase activity.
Tsakiris S,
Giannoulia-Karantana A,
Simintzi I,
Schulpis KH.
Department of Experimental Physiology, Medical School,
University of Athens, P.O. Box 65257, GR-154 01 Athens, Greece.

Stylianos Tsakiris. stsakir@...,

Giannoulia-Karantana A. First Department of Pediatrics,
Aghia Sophia Children's Hospital, University of Athens, Greece.

Kleopatra H. Schulpis, MD, PhD. Institute of Child Health,
Aghia Sophia Children's Hospital, GR-11527 Athens (Greece)
Tel. +30 1 7708291, Fax +30 1 7700111 inchildh@...,

[ Papoutsakis T. tina.papoutsakis@...,

Papadopoulos G. Department of Biochemistry and Biotechnology,
University of Thessaly, Ploutonos 26, 41221 Larisa, Greece
papg@..., ]

Abstract:

Studies have implicated aspartame (ASP) with neurological problems.
The aim of this study was to evaluate acetylcholinesterase (AChE)
activity in human erythrocyte [red blood cell] membranes
after incubation with the sum of ASP metabolites,
phenylalanine (Phe),
methanol (met) and
aspartic acid (aspt),
or with each one separately.

Erythrocyte [human red blood cell] membranes were obtained from
12 healthy individuals and were incubated with ASP hydrolysis
products for 1h at 37 degrees C.
AChE was measured spectrophotometrically.

Incubation of membranes with ASP metabolites corresponding
with 34 mg/kg, 150 mg/kg or 200 mg/kg of ASP consumption
resulted in an enzyme activity reduction by -33%, -41%, and -57%,
respectively.

Met concentrations 0.14 mM, 0.60 mM, and 0.80 mM decreased
the enzyme activity by -20%, -32% or -40%, respectively.

Aspt concentrations 2.80 mM, 7.60 mM or 10.0 mM inhibited
membrane AChE acitivity by -20%, -35%, and -47%, respectively.

Phe concentrations 0.14 mM, 0.35 mM or 0.50 mM reduced the
enzyme activity by -11%, -33%, and -35%, respectively.

Aspt or Phe concentrations 0.82 mM or 0.07 mM, respectively,
did not alter the membrane AChE activity.

It is concluded that low concentrations of ASP metabolites had
no effect on the membrane enzyme activity,
whereas high or toxic concentrations partially or remarkably
decreased the membrane AChE activity, respectively.
Additionally, neurological symptoms, including learning and memory
processes, may be related to the high or toxic concentrations of the
sweetener metabolites. PMID: 16129618


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


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

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

This study demonstrated that chronic aspartame consumption in rats
can lead to altered T-maze performance and increased muscarinic
cholinergic receptor densities in certain brain regions.
Control and treated rats were trained in a T-maze to a particular side
and then periodically tested to see how well
they retained the learned response.
Rats that had received aspartame (250 mg/kg/day)
in the drinking water for 3 or 4 months showed a significant increase
in time to reach the reward in the T-maze,
suggesting a possible effect on memory due to the artificial sweetener.
Using [(3)H]quinuclidinyl benzilate (QNB) (1 nM) to label muscarinic
cholinergic receptors and atropine (10(-6) M) to determine nonspecific
binding in whole-brain preparations,
aspartame-treated rats showed a 31 % increase in receptor numbers
when compared to controls.
In aspartame-treated rats, there was a significant increase in muscarinic
receptor densities in the
frontal cortex, midcortex, posterior cortex, hippocampus, hypothalamus
and cerebellum of
80 %, 60 %, 61 %, 65 %, 66 % and 60 %, respectively.
The midbrain was the only area where preparations from
aspartame-treated rats showed a significant increase
in Na(+),K(+)-ATPase activity.
It can be concluded from these data that long-term consumption
of aspartame can affect T-maze performance in rats and alter
receptor densities or enzymes in brain. PMID: 15159141
//////////////////////////////////////////////////////////


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

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

http://groups.yahoo.com/group/aspartameNM/messages
group with 76 members, 1,444 posts in a public, searchable archive
http://RMForAll.blogspot.com

http://groups.yahoo.com/group/aspartameNMmessage/1443
Safe Food Campaign wants ban on aspartame in schools in New Zealand:
Murray 2007.06.21

http://groups.yahoo.com/group/aspartameNM/message/1442
Wellington, NZ lady, 25, free by 24 hours of severe muscle cramps (5
months) after quitting 4-8 packs daily aspartame chewing gum (past few
years): Murray 2007.06.20

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

www.ehponline.org/members/2007/10271/10271.pdf free full text 24
pages

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

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

http://groups.yahoo.com/group/aspartameNM/message/1426
ASDA (unit of Wal-Mart Stores WMT.N) and Marks & Spencer
will join Tesco and also Sainsbury to ban and limit aspartame,
MSG, artificial flavors dyes preservatives additives, trans fats,
salt "nasties" to protect kids from ADHD: leading UK media:
Murray 2007.05.15

http://groups.yahoo.com/group/aspartameNM/message/1271
combining aspartame and quinoline yellow, or MSG and
brilliant blue, harms nerve cells, eminent C. Vyvyan
Howard et al, 2005 education.guardian.co.uk,
Felicity Lawrence: Murray 2005.12.21

http://groups.yahoo.com/group/aspartameNM/message/1277
50% UK baby food is now organic -- aspartame or MSG
with food dyes harm nerve cells, CV Howard 3 year study
funded by Lizzy Vann, CEO, Organix Brands,
Children's Food Advisory Service: Murray 2006.01.13

formaldehyde as a potent unexamined cofactor in cancer research --
sources include methanol, dark wines and liquors, aspartame, wood and
tobacco smoke: IARC Monographs on the Evaluation of Carcinogenic Risks
to Humans implicate formaldehyde in #88 and alcohol drinks in #96:
some related abstracts: Murray 2007.05.15
http://groups.yahoo.com/group/aspartameNM/message/1417

aspartame (methanol, formaldehyde) toxicity research summary:
Rich Murray 2007.06.16
http://groups.yahoo.com/group/aspartameNM/message/1404

One liter aspartame diet soda, about 3 12-oz cans,
gives 61.5 mg methanol,
so if 30% is turned into formaldehyde, the formaldehyde
dose of 18.5 mg is 37 times the recent EPA limit of
0.5 mg per liter daily drinking water for a 10-kg child:
www.epa.gov/teach/chem_summ/Formaldehyde_summary.pdf
2007.01.05 [ does not discuss formaldehyde from methanol
or aspartame ]
http://www.epa.gov/teach/teachsurvey.html comments
teach@...

http://groups.yahoo.com/group/aspartameNM/message/1340
aspartame groups and books: updated research review of
2004.07.16: Murray 2006.05.11

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

Dark wines and liquors, as well as aspartame, provide
similar levels of methanol, above 120 mg daily, for
long-term heavy users, 2 L daily, about 6 cans.

Within hours, methanol is inevitably largely turned into
formaldehyde, and thence largely into formic acid -- the
major causes of the dreaded symptoms of "next morning"
hangover.

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 18.5 times the USA EPA limit for daily formaldehyde in
drinking water, 2.0 mg in 2 L average daily drinking water.

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

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
//////////////////////////////////////////////////////////