| Glyoxal is directly genotoxic in vitro in bacterial and mammalian
cells. In vivo tests show various findings. A detailed overview of genotoxicity tests
in bacterial test systems is published in the source document (BUA, 1997).
In the Salmonella microsomal assay, glyoxal (test substance 30-40%
glyoxal) was a direct mutagen in strains TA 100, TA 102, TA 104, and TA 2638,
with a weaker response in the presence of a metabolic activation system (BUA,
1997). A direct genotoxic activity of glyoxal was further evident in the
L-arabinose resistance assay with S. typhimurium BA9 and BA13 (Ruiz-Rubio et al., 1985; Ariza et al., 1988) and in the SOS chromotest with E. coli PQ37 (von der Hude et al., 1988).
Furthermore, DNA repair tests yielded positive responses in
both the presence and absence of metabolic activation systems, as in the
SOS umu-test with S.
typhimurium TA 1535/pSK 1002 (Ono et al., 1991a,b), in the rec-assay with Bacillus subtilis (also with metabolic activation; Matsui et al., 1989), and in the differential DNA repair test with E. coli K-12/343/636
uvrB+/recA+ and K-12/343/591
uvrB-/recA- (Hellmer & Bolcsfoldi, 1992a). When the latter test was performed
as a host-mediated assay in mice, with oral application of 570 or 1700
mg glyoxal/kg body weight and intravenous application of the bacteria,
a genotoxic effect was not demonstrable in bacteria isolated from blood,
liver, lungs, kidneys, or testicles (Hellmer & Bolcsfoldi, 1992b),
which may be explained by the high reactivity of glyoxal ? for example,
with proteins . In Saccharomyces cerevisiae D61.M, induction of mitotic recombinations pointed to reaction of glyoxal
with DNA, whereas modification of proteins was indicated by chromosome
losses (in the presence of propionitrile, which is a strong inducer of
chromosomal malsegregation), suggesting interference of glyoxal with microtubular
function (Zimmermann & Mohr, 1992).
With E. coli WP2 uvrA, in both the absence and presence of metabolic activation, negative
test results were found in the standard plate incorporation assay (Hoechst
AG, 1984f), whereas an insufficiently documented preincubation assay reported
positive test results (Kato et al., 1989). Ueno et al. (1991b) investigated the characteristics of mutagenicity by glyoxal (particularly
a possible role of active oxygen species) in S. typhimurium TA 100 and TA
104. The scavengers of singlet oxygen almost completely suppressed the mutagenic
action of glyoxal.
A direct genotoxic action of glyoxal was established in a
variety of tests with mammalian cells without metabolic activation (see
BUA, 1997): in a mutagenicity test with mouse lymphoma cells (TK assay)
(Wangenheim & Bolcsfoldi, 1988), in chromosomal aberration tests with
Chinese hamster ovary (CHO) cells (NOTOX, 1986) and V79 cells (Nishi et al., 1989), and in tests for the induction of unscheduled DNA synthesis in
TC-SV40 cells of Syrian hamster (Cornago et al., 1989), for the induction of sister chromatid exchanges in CHO cells and
human lymphocytes, for the induction of endoreduplication in CHO cells
(Tucker et al., 1989), and for the induction of DNA strand breaks in mouse lymphoma cells
(Garberg et al., 1988). In primary rat hepatocytes, glyoxal induced DNA single strand
breaks but no DNA cross-links (Ueno et al., 1991c).
DNA damage was further demonstrated in the comet assay with
TK6 human lymphoblastoid cells by the induction of concentration-dependent
increases of tail moment and tail length (Henderson et al., 1998). Primary rat hepatocytes exposed to glyoxal at higher concentrations
(0.5-10 mg/ml) produced different concentration-dependent types of DNA
damage. Tail moment and the formation of comets with head and tail (indicative
of DNA strand breakage) decreased with increasing glyoxal concentration,
whereas circular DNA spots with highly condensed areas increasingly appeared
at the mid- and high concentrations. Among 100 tested substances, this
damage was shown to be specific for certain aldehydes and was attributed
to their DNA cross-linking activity (Kuchenmeister et al., 1998). In cultures of human umbilical vein endothelial cells, addition
of 100 Κg glyoxal/ml caused a significant increase of formamidopyrimidine
N-glycosylase (FPG)-sensitive sites (measured by the comet assay) in the
absence of increased intracellular levels of hydroperoxides. FPG repairs
oxidative DNA damage and abasic sites and further was supposed to repair
guanine-glyoxal adducts (Shimoi et al., 2001).
A significantly increased rate of sex-linked recessive lethals
reported in Drosophila melanogaster in preliminary experiments (Mazar Barnett & Munoz, 1969) was not confirmed
in later assays, showing the absence of any genotoxic effect in assays
for sex-linked recessive lethals in mature sperm and in the earlier stages
of spermatogenesis, as well as in assays for clastogenic activity in mature
sperm (reciprocal translocation, dominant lethal, and chromosome loss).
However, from the increase of radiation-induced clastogenic effects after
pretreatment with glyoxal, it was concluded that glyoxal came in contact
with the target cells. The possibility of detoxifying mechanisms for glyoxal
or of an efficient repair of glyoxal-induced damage in Drosophila was discussed (Mazar Barnett & Munoz, 1989).
No clastogenic activity was found in a micronucleus assay
in mouse bone marrow (Societe Francaise Hoechst, 1986; no further data
available).
Glyoxal was demonstrated to be genotoxic at the site of application
after administration by gastric intubation. In the pyloric mucosa of male
Fischer 344 rats, both significantly increased unscheduled DNA synthesis
and DNA single strand breaks were induced at dosages of 400-500 mg/kg body
weight within 2 h. Cytotoxicity was not reported (Furihata et al., 1985, 1988, 1989; Furihata & Matsushima, 1989). In contrast, in
rat hepatocytes, a test for unscheduled DNA synthesis was negative (CCR,
1992). Glyoxal has also been shown to cause DNA strand breaks in rat hepatocytes
2-9 h after a single oral exposure to 200-1000 mg glyoxal/kg body weight
(Ueno et al., 1991b). Single strand breaks were also detected in livers of rats within
2 h following a single oral exposure at 200-1000 mg glyoxal/kg body weight.
The frequency of breaks reached a maximum after 9 h of exposure. Hardly
any DNA lesions were detected in other tissues following exposure to 1000
mg glyoxal/kg body weight. Glyoxal causes DNA single strand breaks in rat
hepatocytes following in vitro and in vivo
exposure (Ueno et al., 1991c).
Cell transformation assays in C3H/10T? cells with three different
commercial products of glyoxal (test concentrations from 0.0013 to 0.195
Κl/ml) yielded negative test results (Mason 1980a,b,c).
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