DNA Damage from Sucralose, Cyclamate and Saccharin

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Sugar feeds cancer.

 

http://health.groups.yahoo.com/group/aspartameNM/message/935
comet assay finds DNA damage from sucralose, cyclamate, saccharin in mice: Sasaki YF & Tsuda S  Aug 2002: Murray 1.1.3 rmforall:
FDA Docket 02P-0317 Recall Aspartame as a Neurotoxic Drug

Please post this to the FDA Dockets website.

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

Sasaki YF & Tsuda S (2002):

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

.....four sweeteners (sodium cyclamate, saccharin, sodium saccharin, and
sucralose) also induced DNA damage in gastrointestinal organs....

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

[ Aspartame, neotame, acesulfamine-K, and stevia should be tested
immediately. ]

[ See:  http://health.groups.yahoo.com/group/aspartameNM/message/934
24 recent formaldehyde toxicity [Comet assay] reports:
Murray 12.31.2 rmforall:
FDA Docket 02P-0317 Recall Aspartame as a Neurotoxic Drug  ]

The Single Cell Gel Assay is able to detect single-strand and
double-strand DNA breaks in individual eukaryotic cells; requires small
numbers of cells (<20,000 per sample); can detect DNA damage from low
levels of toxic or physical insults; and is rapid, simple and efficient.
In this assay, cells are treated with the agent of interest, embedded in
agarose on a histological slide, the cell membranes are lysed, and the
slides are placed in an electric field. If the DNA has single or
double-strand breaks, it will flow out of the cells and move toward the
anode, causing the cell and its DNA to resemble a comet. The more DNA
released from the cell, the greater the DNA damage. A computerized
imaging system is used to score and measure the comets.

http://cometassay.com/  Comet Assay Interest Group

http://www.ems-us.org/   Environmental Mutagen Society
http://www.geocities.com/cometassay/related_links.htm
DNA repair Interest Group  about a thousand members

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

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

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

Of all the additives, dyes were the most genotoxic. Amaranth, Allura
Red, New Coccine, Tartrazine, Erythrosine, Phloxine, and Rose Bengal
induced dose-related DNA damage in the glandular stomach, colon, and/or
urinary bladder.

All seven dyes induced DNA damage in the gastrointestinal organs at a
low dose (10 or 100mg/kg).

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

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

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

Toxicol Sci 2001 May;61(1):92-9
DNA damage induced by red food dyes orally administered to pregnant
and male mice.
Tsuda S, Murakami M, Matsusaka N, Kano K, Taniguchi K, Sasaki YF.

Laboratory of Veterinary Public Health, Department of Veterinary
Medicine, Faculty of Agriculture, Iwate University, Ueda 3-18-8,
Morioka, Iwate 020-8550, Japan.  s.tsuda@iwate-u.ac.jp

We determined the genotoxicity of synthetic red tar dyes currently used
as food color additives in many countries, including JAPAN: For the
preliminary assessment, we treated groups of 4 pregnant mice
(gestational day 11) once orally at the limit dose (2000 mg/kg) of
amaranth (food red No. 2), allura red (food red No. 40), or acid red
(food red No. 106), and we sampled brain, lung, liver, kidney, glandular
stomach, colon, urinary bladder, and embryo 3, 6, and 24 h after
treatment.

We used the comet (alkaline single cell gel electrophoresis) assay to
measure DNA damage. The assay was positive in the colon 3 h after the
administration of amaranth and allura red and weakly positive in the
lung 6 h after the administration of amaranth.

Acid red did not induce DNA damage in any sample at any sampling time.

None of the dyes damaged DNA in other organs or the embryo.

We then tested male mice with amaranth, allura red, and a related
color additive, new coccine (food red No. 18). The 3 dyes induced DNA
damage in the colon starting at 10 mg/kg.

Twenty ml/kg of soaking liquid from commercial red ginger pickles, which
contained 6.5 mg/10 ml of new coccine, induced DNA damage in colon,
glandular stomach, and bladder.

The potencies were compared to those of other rodent carcinogens. The
rodent hepatocarcinogen p-dimethylaminoazobenzene induced colon DNA
damage at 1 mg/kg, whereas it damaged liver DNA only at 500 mg/kg.

Although 1 mg/kg of N-nitrosodimethylamine induced DNA damage in liver
and bladder, it did not induce colon DNA damage. N-nitrosodiethylamine
at 14 mg/kg did not induce DNA damage in any organs examined. Because
the 3 azo additives we examined induced colon DNA damage at a very low
dose, more extensive assessment of azo additives is warranted.
PMID: 11294979

Crit Rev Toxicol 2000 Nov;30(6):629-799
The comet assay with multiple mouse organs: comparison of comet assay
results and carcinogenicity with 208 chemicals selected from the IARC
monographs and U.S. NTP Carcinogenicity Database.
Sasaki YF, Sekihashi K, Izumiyama F, Nishidate E, Saga A, Ishida K,
Tsuda S.
Laboratory of Genotoxicity, Faculty of Chemical and Biological
Engineering, Hachinohe National College of Technology, Hachinohe,
Aomori, Japan. yfsasaki-c@hachinohe-ct.ac.jp

The comet assay is a microgel electrophoresis technique for detecting
DNA damage at the level of the single cell. When this technique is
applied to detect genotoxicity in experimental animals, the most
important advantage is that DNA lesions can be measured in any organ,
regardless of the extent of mitotic activity.

The purpose of this article is to summarize the in vivo genotoxicity in
eight organs of the mouse of 208 chemicals selected from International
Agency for Research on Cancer (IARC) Groups 1, 2A, 2B, 3, and 4, and
from the U.S. National Toxicology Program (NTP) Carcinogenicity
Database, and to discuss the utility of the comet assay in genetic
toxicology.

Alkylating agents, amides, aromatic amines, azo compounds, cyclic nitro
compounds, hydrazines, halides having reactive halogens, and polycyclic
aromatic hydrocarbons were chemicals showing high positive effects in
this assay.

The responses detected reflected the ability of this assay to detect the
fragmentation of DNA molecules produced by DNA single strand breaks
induced chemically and those derived from alkali-labile sites developed
from alkylated  bases and bulky base adducts.

The mouse or rat organs exhibiting increased levels of DNA damage were
not necessarily the target organs for carcinogenicity.

It was rare, in contrast, for the target organs not to show DNA damage.

Therefore, organ-specific genotoxicity was necessary but not sufficient
for the prediction of organ-specific carcinogenicity.

It would be expected that DNA crosslinkers would be difficult to detect
by this assay, because of the resulting inhibition of DNA unwinding. The
proportion of 10 DNA crosslinkers that was positive, however, was high
in the gastrointestinal mucosa, stomach, and colon, but less than 50% in
the liver and lung. It was interesting that the genotoxicity of DNA
crosslinkers could be detected in the gastrointestinal organs even
though the agents were administered intraperitoneally.

Chemical carcinogens can be classified as genotoxic (Ames test-positive)
and putative nongenotoxic (Ames test-negative) carcinogens. The Ames
test is generally used as a first screening method to assess chemical
genotoxicity and has provided extensive information on DNA reactivity.

Out of 208 chemicals studied, 117 are Ames test-positive rodent
carcinogens, 43 are Ames test-negative rodent carcinogens, and 30 are
rodent noncarcinogens (which include both Ames test-positive and
negative noncarcinogens).

High positive response ratio (110/117) for rodent genotoxic       
carcinogens and a high negative response ratio (6/30) for rodent
noncarcinogens were shown in the comet assay.

For Ames test-negative rodent carcinogens, less than 50% were positive
in the comet assay, suggesting that the assay, which detects DNA
lesions, is not suitable for identifying nongenotoxic carcinogens. In
the safety evaluation of chemicals, it is important to demonstrate that
Ames test-positive agents are not genotoxic in vivo.

This assay had a high positive response ratio for rodent genotoxic
carcinogens and a high negative response ratio for rodent genotoxic
noncarcinogens, suggesting that the comet assay can be used to evaluate
the in vivo genotoxicity of in vitro genotoxic chemicals.

For chemicals whose in vivo genotoxicity has been tested in multiple
organs by the comet assay, published data are summarized with
unpublished data and compared with relevant genotoxicity and
carcinogenicity data. Because it is clear that no single test is capable
of detecting all relevant genotoxic agents, the usual approach should be
to carry out a battery of in vitro and in vivo tests for genotoxicity.

The conventional micronucleus test in the hematopoietic system is a
simple method to assess in vivo clastogenicity of chemicals. Its
performance is related to whether a chemical reaches the hematopoietic
system. Among 208 chemicals studied (including 165 rodent carcinogens),
54 rodents carcinogens do not induce micronuclei in mouse hematopoietic
system despite the positive finding with one or two in vitro tests.

Forty-nine of 54 rodent carcinogens that do not induce micronuclei were
positive in the comet assay, suggesting that the comet assay can be used
as a further in vivo test apart from the cytogenetic assays in
hematopoietic cells. In this review, we provide one recommendation for
the in vivo comet assay protocol based on our own data.
Publication Types: Review  Review, Tutorial  PMID: 11145306
************************************************************************

http://health.groups.yahoo.com/group/aspartameNM/message/932
aspartame: methanol, formaldehyde, formic acid toxicity:
brief review: Murray 12.30.2 rmforall:

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

http://health.groups.yahoo.com/group/aspartameNM/messagesfor 935 posts in a public searchable archive

http://health.groups.yahoo.com/group/aspartameNM/message/910
formaldehyde & formic acid from methanol in aspartame:
Murray: 12.9.2 rmforall

It is certain that high levels of aspartame use, above 2 liters daily
for months and years, must lead to chronic formaldehyde-formic acid
toxicity, since 11% of aspartame (1,120 mg in 2L diet soda, 5.6 12-oz
cans) is 123 mg methanol (wood alcohol), immediately released into the
body after drinking (unlike the large levels of methanol locked up in
molecules inside many fruits), then quickly transformed into
formaldehyde, which in turn becomes formic acid, both of which in
time become carbon dioxide and water-- however, about 30% of the
methanol remains in the body as cumulative durable toxic metabolites of
formaldehyde and formic acid-- 37 mg daily, a gram every month.
If 10% of the methanol is retained as formaldehyde, that would give 12
mg daily formaldehyde accumulation, about 60 times more than the 0.2 mg
from 10% retention of the 2 mg EPA daily limit for formaldehyde in
drinking water.

Bear in mind that the EPA limit for formaldehyde in
drinking water is 1 ppm,
or 2 mg daily for a typical daily consumption of 2 L of water.

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

This long-term low-level chronic toxic exposure leads to typical
patterns of increasingly severe complex symptoms, starting with
headache, fatigue, joint pain, irritability, memory loss, and leading to
vision and eye problems and even seizures. In many cases there is
addiction.  Probably there are immune system disorders, with a
hypersensitivity to these toxins and other chemicals.

Confirming evidence and a general theory are given by Pall (2002):
http://health.groups.yahoo.com/group/aspartameNM/message/909
testable theory of MCS type diseases, vicious cycle of nitric oxide &
peroxynitrite: MSG: formaldehyde-methanol-aspartame:
Martin L. Pall: Murray: 12.9.2 rmforall

http://health.groups.yahoo.com/group/aspartameNM/message/915
formaldehyde toxicity:  Thrasher & Kilburn: Shaham: EPA: Gold: Murray:
Wilson: CIIN: 12.12.2 rmforall

http://health.groups.yahoo.com/group/aspartameNM/message/868
Murray: submit complaints and papers to FDA Docket 02P-0317
by Jan 12  2003: Recall Aspartame as a Neurotoxic Drug 9.20.2 rmforall

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



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