Rapid isolation of carcinogen-bound DNA and RNA by hydroxyapatite chromatography

Beland, Frederick A.; Dooley, Kenneth L.; Casciano, Daniel A.

doi:10.1016/s0021-9673(00)87048-x

Cited by 126 publications

(34 citation statements)

References 28 publications

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“…Care was exercised to ensure that the highest quality phenol (Aldrich Chemical Co., Gold Label) was used. For initial experiments, DNA was isolated by hydroxyapatite chromatography (1).…”

Section: Dna Isolation and Digestionmentioning

confidence: 99%

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Increased 8-Hydroxyguanine Content of Chloroplast DNA from Ozone-Treated Plants

Floyd¹,

West²,

Hogsett³

et al. 1989

Plant Physiol.

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The mechanism of ozone-mediated plant injury is not known but has been postulated to involve oxygen free radicals. Hydroxyl free radicals react with DNA causing formation of many products, one of which is 8-hydroxyguanine. By using high performance liquid chromatography with electrochemical detection, the 8-hydroxy-2'-deoxyguanosine (8-OHdG) content of a DNA enzymatic digest can be sensitively quantitated. Beans (Phaseolus vulgaris L.) and peas (Pisum sativum L.) were treated with an ozone regime that caused acute injury. Chloroplast DNA was obtained from plants harvested either immediately after ozone treatment or 24 hours later. Ozone-exposed plants in general had nearly two-fold higher levels of 8-OHdG as compared to control plants. In (7,9,10). Hydroxyl free radicals react with DNA to yield numerous products; one is guanine, which becomes hydroxylated at the 8-carbon position (10, 16). Following hydroxyl free radical reaction with DNA and its digestion to the nucleoside, 8-OHdG2 (7, 10, 16) can then be directly quantitated by LCED. The detection sensitivity with LCED is 20 fmol (7). With DNA as an indicator of hydroxyl free radical occurrence and LCED methodology, it was possible to ascertain that oxidative damage to DNA had occurred in a living system subjected to oxidative stress (7). Kasai

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“…Care was exercised to ensure that the highest quality phenol (Aldrich Chemical Co., Gold Label) was used. For initial experiments, DNA was isolated by hydroxyapatite chromatography (1).…”

Section: Dna Isolation and Digestionmentioning

confidence: 99%

“…It has been implicated as a key agent which, in part, may be responsible for the tree decline observed in Europe and in parts of the United States and Canada (2, 1 1,19,21,26). Ozone-mediated plant injury has been investigated for some time, yet no clear-cut mechanisms have emerged.…”

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confidence: 99%

Increased 8-Hydroxyguanine Content of Chloroplast DNA from Ozone-Treated Plants

Floyd¹,

West²,

Hogsett³

et al. 1989

Plant Physiol.

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“…DNA adducts are found in many organs after administration of aromatic amines and derivatives, often also in nontarget organs. In target organs, N-acetylarylamine as well as arylamine adducts are found [e.g., from 2-acetylaminofluorene and N-hydroxy-2-acetylaminofluorene in the male rat liver (3,4,7,18,19), and from N-hydroxy-4'-fluoro-4-acetylaminobiphenyl in the rat liver and kidney (20,21)]. However, in other target organs, only arylamine adducts have been found, [e.g., from 4-aminobiphenyl in the dog bladder (22)(23)(24), N-hydroxy-2-acetylaminofluorene in the rat mammary gland (25), and 3,2'-dimethyl-4-aminobiphenyl in rat colon (26)].…”

Section: Introductionmentioning

confidence: 99%

Metabolic activation routes of arylamines and their genotoxic effects.

Meerman

Poll

1994

Environ Health Perspect

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Two different types of DNA adducts are formed from many aromatic amines by bioactivation: N-acetylated and nonacetylated, arylamine DNA adducts. It has become clear from experiments using N-acetyl-2-aminofluorene and 2-aminofluorene adducts to C8 of deoxyguanosine that these two types of adducts may have different effects on DNA structure and DNA replication. We have determined blocking of DNA replication by various other N-acetylarylamine and arylamine deoxyguanosine adducts. It was found that the N-acetyl group in general is required for blocking of DNA replication; the nature of the aromatic moiety seems to be of minor importance. Little information is available on the genotoxic effects of these adducts in mammalian cells in vivo. We have tried to get more insight in this by investigating the clastogenicity, the initiation of preneoplastic cells, and the promotional effects of various aromatic amines from which different ratios of N-acetylarylamine DNA adducts to arylamine DNA adducts are formed in the rat liver. Our results show that formation of N-acetylarylamine adducts to C8 of deoxyguanosine in the liver is correlated with clastogenicity and hepatic promoting effect. Initiation capacities, however, seem to be correlated with formation of nonacetylated, arylamine adducts. Mechanisms by which formation of N-acetylarylamine DNA adducts may generate a promoting effect in the liver are discussed. -Environ Health Perspect 102(Suppl 6): 153-159 (1994)

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“…The antiserum is specific of the acetyl-*In Vitro Pathogenesis Section, Laboratory of Cellular Carcinogenesis and Tumor Promotion, National Cancer Institute, Bethesda, MD 20205. tAuthor to whom reprint requests should be addressed. (N-Ac-AAF) with DNA in vivo, including cultured cells (6)(7)(8)(9) and whole animals (6,(10)(11)(12)(13)(14). We have utilized RIA to quantitate formation and removal of guan-8-yl (C-8) adducts in liver and kidney DNA of male rats fed 2-acetylaminofluorene during a period of 2 months.…”

Section: Introductionmentioning

confidence: 99%

Determination of 2-Acetylaminofluorene Adducts by Immunoassay

Poirier¹,

True²,

Laishes³

1983

Environmental Health Perspectives

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Antisera elicited in rabbits were used in radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) to determine femtomole quantities of deoxyguanosin-8-yl-acetylaminofluorene (dg-8-AAF) and deoxyguanosin-8-yl-aminofluorene (dg-8-AF). These adducts have been monitored in liver and kidney DNA of male Wistar-Furth rats fed 0.02% or 0.04% 2-acetylaminofluorene (2-AAF) either continuously or for a limited time followed by an interval on control diet. After 24 hr of 0.02% 2-AAF feeding, substantial levels of binding (80 fmole/4g DNA) were observed in liver DNA and increased with time, reaching a plateau of approximately 230 fmole/,Ag DNA at 30 days and thereafter. During the first week of continuous feeding about 80% of the total C-8 adducts in the liver DNA were deacetylated (dG-8-AF). By 25-60 days, dG-8-AF represented 97-100% of all C-8 adducts as measured by RIA and confirmed by HPLC. Values for C-8 adduct formation in kidney DNA were severalfold lower than in liver and dG-8-AF represented >90% of C-8 adducts at all times studied.In removal or repair experiments, rats were fed 2-AAF for 3, 7 or 28 days, the 2-AAF diet was discontinued and the liver adducts assayed after intervals on control diet. When dietary 2-AAF administration was for 3 or 7 days, removal of adducts was efficient and almost complete by 28 days on control diet, with preferential retention of dG-8-AF. However, when dietary 2-AAF administration was for 28 days, adduct levels were higher, the repair capacity was saturated and the removal of C-8 adducts was not complete after control diet for a 28-day interval. In a preliminary experiment when[3H]-2-AAF was fed for 3 days, after 25 days of 0.02% 2-AAF, the rates of newly formed adduct formation and removal were similar to those observed for the initial 3 days of 2-AAF feeding. These results demonstrate the predominance and persistence of dG-8-AF in liver and kidney DNA of 2-AAFfed rats and suggest that the repair capacity of the whole rat liver was not diminished after 1 month of 2-AAF feeding.

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Rapid isolation of carcinogen-bound DNA and RNA by hydroxyapatite chromatography

Cited by 126 publications

References 28 publications

Increased 8-Hydroxyguanine Content of Chloroplast DNA from Ozone-Treated Plants

Increased 8-Hydroxyguanine Content of Chloroplast DNA from Ozone-Treated Plants

Metabolic activation routes of arylamines and their genotoxic effects.

Determination of 2-Acetylaminofluorene Adducts by Immunoassay

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