Determination of malondialdehyde-induced DNA damage in human tissues using an immunoslot blot assay

Leuratti, Chiara; Singh, Rajinder; Lagneau, Christian; Farmer, Peter B.; Plastaras, John P.; Marnett, L.J.; Shuker, David E. G.

doi:10.1093/carcin/19.11.1919

Cited by 98 publications

(67 citation statements)

References 15 publications

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“…Extraction of genomic DNA from liver tissue obtained at staging laparoscopy and during surgery, and analysis of M 1 G adduct levels by immunoslotblot was performed as described previously by Leuratti et al (1998) using a primary anti-M 1 G antibody provided by Dr Lawrence Marnett (Vanderbilt University, TN, USA). Discrepancies in the amount of DNA in each slot were corrected for by staining the nitrocellulose filter with propidium iodide and performing UV light densitometry.…”

Section: Analysis Of M 1 G Adduct Levelsmentioning

confidence: 99%

Detection of curcumin and its metabolites in hepatic tissue and portal blood of patients following oral administration

et al. 2004

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Studies in vitro and in animal models of colorectal and hepatocellular cancers suggest that curcumin is an effective chemopreventive agent. In this pilot trial, we investigated whether oral administration of curcumin results in concentrations of the agent in normal and malignant human liver tissue, which are sufficient to elicit pharmacological activity. In total, 12 patients with hepatic metastases from colorectal cancer received 450 -3600 mg of curcumin daily, for 1 week prior to surgery. Levels of curcumin and its metabolites were measured by HPLC in portal and peripheral blood, bile and liver tissue. Curcumin was poorly available, following oral administration, with low nanomolar levels of the parent compound and its glucuronide and sulphate conjugates found in the peripheral or portal circulation. While curcumin was not found in liver tissue, trace levels of products of its metabolic reduction were detected. In patients who had received curcumin, levels of malondialdehyde-DNA (M 1 G) adduct, which reflect oxidative DNA changes, were not decreased in post-treatment normal and malignant liver tissue when compared to pretreatment samples. The results suggest that doses of curcumin required to furnish hepatic levels sufficient to exert pharmacological activity are probably not feasible in humans.

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Section: Analysis Of M 1 G Adduct Levelsmentioning

confidence: 99%

Detection of curcumin and its metabolites in hepatic tissue and portal blood of patients following oral administration

et al. 2004

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“…Significant progress has been made in characterizing carbonyl-containing products of lipid peroxidation (e.g. malondialdehyde, 4-hydroxy-2-alkenals) and the adducts they form with DNA (Leuratti et al, 1998;Chen et al, 1998;Moller and Wallin, 1998;Yi et al, 1998) as well as the contributions of physiologic mediators such as nitric oxide in these reactions . The DNA adducts formed in reactions with carcinogenic aldehydes may produce structurally unique adducts (Hecht et al, 2001); these hold promise as sensitive markers of oral cancer risk associated with tobacco use (Nath et al, 1998).…”

Section: Emerging Issues In Tobacco Carcinogenesis and Dna Adductsmentioning

confidence: 99%

DNA adduct burden and tobacco carcinogenesis

Wiencke

2002

Oncogene

149

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DNA adducts associated with tobacco smoking could provide a marker of biologically effective dose of tobacco carcinogens and improve individual cancer risk prediction. A significant number of clinical and epidemiologic studies have reported associations of increased DNA adduct levels with the occurrence of the prevalent tobacco related cancers including cancer of the lung, head and neck, and bladder. The inducibility of DNA adducts following in vitro treatments using blood lymphocytes also appears to be a risk factor in the development of lung and head and neck cancer. Corroborative evidence pointing to the importance of DNA adducts in tobacco carcinogenesis include numerous studies showing associations of tobacco smoke exposure with the induction of DNA adducts in humans in vivo. Further effort is necessary, however, to more fully characterize the dose -response relationship between smoking and DNA adducts in exposed target and surrogate tissues. The relationship between gene polymorphisms thought to modify tobacco-related cancer risk and DNA adduct levels is complex. Results of some DNA adduct studies (both in vitro and in vivo) appear inconsistent with the epidemiologic findings. This is evident for polymorphisms involving both carcinogen metabolism (e.g. GSTP1) and DNA repair (e.g. XRCC1). Molecular studies of human tumors suggest associations of p53 mutation with DNA adducts and have revealed correlations of DNA adduct levels with somatic alterations (e.g. 3p21 LOH) that are thought to occur at the very earliest stages of tobacco carcinogenesis. More research is needed to assess the relationship between endogenous sources of DNA adducts and tobacco smoke exposure and the relative oncogenic effects of chemically stable versus unstable DNA adducts. Many potentially fruitful new avenues of cancer research are emerging that integrate DNA adduct analyses with assessments of smoking, genetics, diet and ambient air quality. These investigations aim to understand the multifactorial nature of interindividual variability in response to tobacco carcinogens. As these trends continue a variety of innovative study designs and approaches will become important in human populations.

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“…A human DNA QC sample (Boehringer Mannheim) was included in all assays for M 1 dG and the results rejected if the QC had a SD of > 20% for the triplicate analyses or the QC result was > 2 SD from the mean of all the QC results (2.00 adducts per 10 7 normal bases +/-sd 1.27). The limit of detection was 0.2 adducts per 10 7 normal nucleotides (15).…”

Section: Analysis Of Blood Samplesmentioning

confidence: 98%

“…Two highly modified M 1 dG-DNA standards were prepared independently in different laboratories as previously described by treating CT-DNA with malondialdehyde (MDA) (15).…”

Section: Preparation Of M 1 Dg-dna Standardsmentioning

confidence: 99%

Optimizing immunoslot blot assays and application to low DNA adduct levels using an amplification approach

Moore

Xeniou

Zeng

et al. 2010

Analytical Biochemistry

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Immunoslot blot assays have been used for the analysis of many DNA adducts but problems are frequently encountered in achieving reproducible results. Each step of the assay has been systematically examined and it was found that the major problems are in the DNA fragmentation step and the use of the manifold apparatus. Optimisation was performed upon both the malondialdehyde-deoxyguanosine adduct (M 1 dG) and the O 6 -carboxymethyldeoxyguanosine adduct (O 6 CMdG) to demonstrate the applicability to other DNA adducts.Blood samples from the EPIC study (n = 162) were analysed for M 1 dG adducts and the data showed no correlation with adduct levels in other tissues indicating that the EPIC blood samples were not useful for studying M 1 dG adducts. Blood samples from a processed meat vs vegetarian diet intervention (n = 6) were analysed for O 6 CMdG and many were below the limit of detection. The reduction of background adduct levels in standard DNA was investigated using chemical and whole-genome amplification approaches. The latter gave a sensitivity improvement of 2.6 adducts per 10 7 nucleotides for the analysis of O 6 CMdG.Subsequent reanalysis for O 6 CMdG showed a weakly significant increase in O 6 CMdG on the processed meat diet compared with the vegetarian diet, demonstrating that further studies are warranted.

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Determination of malondialdehyde-induced DNA damage in human tissues using an immunoslot blot assay

Cited by 98 publications

References 15 publications

Detection of curcumin and its metabolites in hepatic tissue and portal blood of patients following oral administration

Detection of curcumin and its metabolites in hepatic tissue and portal blood of patients following oral administration

DNA adduct burden and tobacco carcinogenesis

Optimizing immunoslot blot assays and application to low DNA adduct levels using an amplification approach

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