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  1. #1
    Join Date
    Jun 1999
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    Santa Fe, NM 87505, USA
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    http://groups.yahoo.com/group/aspartameNM/message/961
    genotoxins, Comet assay in mice: Ace-K, stevia fine; aspartame poor;
    sucralose, cyclamate, saccharin bad: Y.F. Sasaki Aug 2002:
    Murray 1.27.3 rmforall

    Jan 27 2003

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

    Summary of Review by Rich Murray: This study tests 39 common food
    additives for DNA damage, comparing a control group of 4 mice against
    test groups of 4 mice each, killed 3 hr and 24 hr after oral
    ingestion of up to 2000 mg/kg.

    However, there are only 21 unique control groups, with widely varying
    values. By using the averages for all 21 control groups to make
    comparison with the groups exposed to the food additives, it is easy
    to see that many additives cause about 140% to about 180% to over 300%
    of the averages of all control groups for the 8 organs measured. By
    using more mice, statistical significance may be easily proved for most
    of these easily noticable high values, which are not significant for
    just 4 mice.

    Mutat Res 2002 Aug 26; 519(1-2): 103-19
    The comet assay with 8 mouse organs: results with 39 currently used food
    additives. Yu F. Sasaki
    Sasaki YF, Kawaguchi S, Kamaya A, Oh****a 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 (2000 mg/kg) and performed the comet
    assay on the glandular stomach, colon, liver, kidney, urinary bladder,
    lung, brain, and bone marrow 3 and 24 h after treatment.

    Of all the additives, dyes were the most genotoxic. Amaranth, Allura
    Red, New Coccine, Tartrazine, Erythrosine, Phloxine, and Rose Bengal
    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

    Also tested were acesulfame K, aspartame, stevia, and glycyrrhizin--
    which all came out nonsignificant, while, as the abstract mentions,
    sodium cyclamate had 4, saccharin 3, sucralose 3,
    and sodium saccharin 5 significant results.

    Each test condition had just 4 mice, and, according to the text, each
    additive had its own control group of 4 mice. However, there are only
    21 unique sets of control groups, with 8 sets used once, 10 sets used
    twice, 2 sets used 3 times, and 1 set used 4 times, a total of 38 food
    additives listed [Sodium erythorbic acid was left out of Table 2, while
    mentioned in the report 3 times,
    "...erythorbic acid and its sodium salt
    did not increase DNA damage in any of the organs studied."].

    Aspartame was assigned the control group that had the highest levels of
    Migration of damaged nuclear DNA for Liver and Bladder, and the second
    highest for Brain. The same control group was used for the xanthene
    dye, erythrosinc, which had Migration as high as 42.4+-2.17 um
    [micro-meter], measured on 50 nuclei from stomach cells, 3 hours
    after ingestion. So, the high control groups values had no effect on
    the statistical analysis for erythrosinc.

    The available range of the 21 control groups ranged for the Liver from
    1.1 to 3.6 um. For aspartame, the Liver Migration, the average length
    of the "comet" tail of damaged, broken DNA pulled out of 50 Liver cell
    nuclei by an electric field for 15 minutes, was, average of 4 mice:

    control value used 3.59+-0.50 um [1.1 to 3.6 range in 21 controls]
    2000 mg/kg 3 hr 3.26+-0.16 um
    2000 mg/kg 24 hr 0.57+-0.22 um

    The 3 hr aspartame test value was about the same as the control value.
    This may be discordant with the Trocho (1998) findings that rats given
    200 mg/kg oral doses of aspartame for 11 days, about the same total
    dose, had accumulation of formaldehyde adducts, bound to DNA, RNA, and
    proteins, in liver, kidneys, brain, retinas,
    and other tissues, at about
    the same total dose, spread over 11 days.

    Appying the lowest available control group liver level 1.06+-0.12 um
    would make the aspartame level of 3.26+-0.16 um significant.

    We should keep in mind that toxicity in humans involves many vulnerable
    groups, years of daily use, often evolution of hypersensitivity, and
    complex interactions with a multitude of foods,
    additives, other toxins, and foods.

    Some of the dye data was earlier published in Tsuda (2001):
    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

    http://groups.yahoo.com/group/aspartameNM/message/935
    comet assay finds DNA damage from sucralose, cyclamate, saccharin in
    mice: Sasaki YF & Tsuda S Aug 2002: Murray 1.1.3 rmforall

    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.
    The Comet assay is not FDA approved as a human medical test, so it is
    not covered by insurance. It is used in many human research studies.

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

    http://www.geocities.com/cometassay/pioneers.htm

    http://www.geocities.com/cometassay/related_links.htm

    http://www.ems-us.org/ Environmental Mutagen Society

    http://www.geocities.com/cometassay/related_links.htm
    DNA repair Interest Group about a thousand members

    http://www.ils-inc.com/dna_damage_assessment.html
    Integrated Laboratory Systems Comet assays for $155-300

    http://www.mdbiotechinc.com/ MD Biotech, Inc.
    Comet assays on four 10 ml blood samples for $800
    ************************************************** *********************

    http://groups.yahoo.com/group/aspartameNM/message/959
    aspartame: methanol, formaldehyde, formic acid toxicity:
    brief review: Murray 2.21.3 rmforall

    http://groups.yahoo.com/group/aspartameNM/messages
    for 968 posts in a public searchable archive

    http://groups.yahoo.com/group/aspartame/ 615 member group

    http://www.holisticmed.com/aspartame...2-response.htm
    Mark Gold exhaustively critiques European Commission Scientific
    Committee on Food re aspartame (12.4.2): 59 pages, 230 references

    http://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://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://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://groups.yahoo.com/group/aspartameNM/message/946
    Functional Therapeutics in Neurodegenerative Disease Part 1/2:
    Perlmutter 7.15.99: Murray 1.10.3 rmforall

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

    http://groups.yahoo.com/group/aspartameNM/message/934
    24 recent formaldehyde toxicity [Comet assay] reports:
    Murray 12.31.2 rmforall

    http://groups.yahoo.com/group/aspartameNM/message/935
    comet assay finds DNA damage from sucralose, cyclamate, saccharin in
    mice: Sasaki YF & Tsuda S Aug 2002: Murray 1.1.3 rmforall

    http://groups.yahoo.com/group/aspartameNM/message/939
    aspartame (aspartic acid, phenylalanine) binding to DNA:
    Karikas July 1998: Murray 1.5.3 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@atlas.uoa.gr

    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

    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://groups.yahoo.com/group/aspartameNM/message/925
    aspartame puts formaldehyde adducts into tissues, Part 1/2
    full text, Trocho & Alemany 6.26.98: Murray 12.22.2 rmforall

    http://groups.yahoo.com/group/aspartameNM/message/926
    aspartame puts formaldehyde adducts into tissues, Part 2/2
    full text, Trocho & Alemany 6.26.98: Murray 12.22.2 rmforall

    http://ww.presidiotex.com/barcelona/index.html
    Trocho C, Pardo R, Rafecas I, Virgili J, Remesar X,
    Fernandez-Lopez JA, Alemany M ["Trok-ho"]
    Formaldehyde derived from dietary aspartame binds to tissue
    components in vivo. Life Sci 1998 Jun 26; 63(5): 337-49.
    Departament de Bioquimica i Biologia Molecular, Facultat de Biologia,
    Universitat de Barcelona, Spain.
    http://www.presidiotex.com/barcelona/index.html
    Maria Alemany, PhD (male) alemany@porthos.bio.ub.es

    http://groups.yahoo.com/group/aspartameNM/message/864
    Murray: Butchko, Tephly, McMartin: Alemany: aspartame formaldehyde
    adducts in rats 9.8.2 rmforall
    Prof. Alemany vigorously affirms the validity of the Trocho study
    against criticism:
    Butchko, HH et al [24 authors], Aspartame: review of safety.
    Regul. Toxicol. Pharmacol. 2002 April 1; 35 (2 Pt 2): S1-93, review
    available for $35, [an industry paid organ]. Butchko:
    "When all the research on aspartame, including evaluations in both the
    premarketing and postmarketing periods, is examined as a whole, it is
    clear that aspartame is safe, and there are no unresolved questions
    regarding its safety under conditions of intended use."
    [They repeatedly pass on the ageless industry deceit that the methanol
    in fruits and vegetables is as as biochemically available as that in
    aspartame-- see the 1984 rebuttal by Monte.]

    http://groups.yahoo.com/group/aspartameNM/message/911
    RTP ties to industry criticized by CSPI: Murray: 12.9.2 rmforall
    ************************************************** *********************
    Send blank post to: <br />aspartameNM-subscribe@onelist.com to join<br />free,open, list with searchable archives for toxicity issues.<br />Richard \"Rich\" T. Murray Room For All 1943 Otowi Road Santa Fe, NM 87505<br />rmforall@comcast.net 505-501-2298

  2. #2
    Join Date
    Dec 1999
    Location
    Steuben County New York State
    Posts
    1,753

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    test post testing
    ."Have a f***ing beer!" Pete Townshend 12-12-12

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