Environmental and Molecular Mutagenesis - Legacy Tobacco ...

600
MONOCLONAL ANTIBOSY IMMUNOASSAYS FOR COOKING-INDUCED MEAT
MUTAGENS .
Martin Vanderlaan, Bruce Watkiss ; Mona Hwang, Mark Knize and James Felton. Biomedical
Sciences Division, Lawrence Livermore National Laboratory, Livermore, CA 94550
Fifteen monoclonal antibodies have been produced that recognize five ne (AIA)
mutagens formed in cooked meats (IQ, MeIQ, MeIQx, DiMeIQx, and Fhlp). These antibodies are
being used to quatttifiy individual AIAs in cooked meats, to identify inunutachemically crosrteac6ve
new AIAs, and to quan c'fy AIAs and their metabolites in human urine . Extracts of well-done cooked
beef were separated on HPLC and competition enzyme linked ittununoaorbent assays (ELISAs) were
conducted on each HPLC fraction using the panel of-antibodies . The plot of intmunochemical activity
vs . HPLC retention time forms an "immunogram," similar to the mone familiar "mutagnm ' plot of
mutagenic activity vs . retention time. Each antibody shows a unique immunogram . Peaks in
inununochemica) activity can be associated with known AIA tnutagani, and withpteviously
unidentified chemicals which immunochemically cross-neact with the antibodies . These findings
suggest that the known AlAs are part of a larger family of immunochemically similar compounds
produced by cooking . Immuno-affinity chromatography is being used to isoLte and identify these
new AIA-like compounds . The antibodies provide a structure-based means of identifying new AIAs
that complements the previously used bacterial mutageatc activity-based methods . Analysis of urine
from people on vegetarian and well-done beef diets s6ows both mutagenic and intmunochemicallypositive
material is associated with the diet high in AIAs .
Research supported by NCIgrants ROl CA40811-03 and CA48446-02 and performed at LLNL under
contract W-7405-ENG-48 with the Department of Energy .
601
MUTAGENICITY OF DRINKING WATERS IN FINLAND . T . Vartiainen, National
Public Health Institute, Dept . Environm . Hyg . Tox . P .O .Box 95, 70701
Kuopio, Finland
Mutagenic activities of drinking and raw waters from the major
waterworks, covering about 60% of all tap water distributed in Finland,
were tested with the Ames Salmonella tvphimurium assay using tester
strains TA100, TA98, and TA97 . The highest yield of organic mytagenic
impurities from these waters was obtained from acidic water samples by
continuous extraction with diethyl ether or by XAD adsorption . Mutagenic
activity of drinking water was dependent on the amount of organic
matter (and ammonia) present in water and thrA quality and quantity of
disinfectant used ; mutagenic activity could be approximated by an equation
of the type A(1-e-kO), where A and k are constants and c is a parameter
that depends on total organic carbon, the chlorine dose, and the
ammonia concentration . The main mutagens in humus-rich chlorinated
drinking water were acidic polar compounds detectable without metabolic
activation . The highly mutagenic compound 3-chloro-4-(dichloromethyl)-
5-hydroxy-2(5H)-furanone (MX) and its geometric isomer E-2-chloro-3-
(dichloromethyl)-4-oxo-butenoic acid (E-MX) were detected in all extracts
that exhibited mutagenicity . The concentrations of MX in the mutagenic
drinking water concentrates ranged from 5 to 67 ng/l and this
compound was responsible for 15 - 57% (average 33%) of the observed mutagenicity
. Linear correlations were observed between mutagenic activity
in TA100 and concentrations of MX and E-MX .
602
ENHANCEMENT OF BLEOMYCIN TOXICITY AND MUTATION AT THE APRT LOCUS IN CHO CELLS
M .L . Veigl, J . Bhatt and W .D . Sedwick, Dept . of Medicine, Case Western Reserve
Univ . Cleveland, OH 44106
In drug combinations causing greater than 95% toxicity for CHO cells, the
calmodulin antagonist, N-(4-aminobutyl)-5-chloro-2-naphthalene sulfonamide
(w13), is synergistic with bleomycin in clonogenic assays as analyzed by the
method of Chou and Talalay (Adv . Enz . Reg ., 22, 27-55, 1984) . At concentrations
causing less toxicity, however, this drug combination interacts in an
antagonistic manner . It has been suggested that calmodulin antagonists may
interfere with DNA repair processes responding to bleomycin-induced DNA damage
. In these experiments, the effect of W13 on mutation caused by bleomycin
was investigated by analyzing the impact of this drug combination on
mutation in the adenine phosphoribosyl transferase gene (aprt) in cells that
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1989 EMS Abstracts 207
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