scholarly journals Effect of 2-acetylaminofluorene and its genotoxic metabolites on DNA adduct formation and DNA damage in 3D reconstructed human skin tissue models

Mutagenesis ◽  
2019 ◽  
Author(s):  
Thomas R Downs ◽  
Volker M Arlt ◽  
Brenda C Barnett ◽  
Ryan Posgai ◽  
Stefan Pfuhler
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Human Skin ◽  
Adduct Formation ◽  
Dna Adduct ◽  
Strong Increase ◽  
Multiple Exposures ◽  
Multiple Treatments ◽  
Reconstructed Human Skin

Abstract In vitro genotoxicity assays utilising human skin models are becoming important tools for the safety assessment of chemicals whose primary exposure is via the dermal route. In order to explore metabolic competency and inducibility of CYP450 activating enzymes, 3D reconstructed human skin tissues were topically treated with 2-acetylaminofluorene (2-AAF) and its genotoxic metabolites, N-hydroxy-2-acetylaminofluorene (N-OH-2-AAF) and N-hydroxy-2-aminofluorene (N-OH-2-AF), which primarily cause DNA damage by forming DNA adducts. 2-AAF did not increase DNA damage measured in the reconstructed skin micronucleus (RSMN) assay when administered in multiple applications at 24 h intervals but was detected in the skin comet assay in the presence of the DNA polymerase inhibitor aphidicolin (APC). Similarly, no increase was found with N-OH-2-AAF in the RSMN assay after multiple treatments whereas a single 3 h exposure to N-OH-2-AAF caused a large dose-related increase in the skin comet assay. A significant increase in the RSMN assay was only obtained with the highly reactive N-OH-2-AF metabolite after multiple treatments over 72 h, whereas N-OH-2-AF caused a strong increase after a single 3 h exposure in the skin comet assay. In support of these results, DNA adduct formation, measured by the 32P-postlabelling assay, was examined. Adduct levels after 2-AAF treatment for 3 h were minimal but increased >10-fold after multiple exposures over 48 h, suggesting that enzyme(s) that metabolise 2-AAF are induced in the skin models. As expected, a single 3 h exposure to N-OH-2-AAF and N-OH-2-AF resulted in adduct levels that were at least 10-fold greater than those after multiple exposures to 2-AAF despite ~100-fold lower tested concentrations. Our results demonstrate that DNA damage caused by 2-AAF metabolites is more efficiently detected in the skin comet assay than the RSMN assay and after multiple exposures and enzyme induction, 2-AAF-induced DNA damage can be detected in the APC-modified comet assay.

Molecular Dosimetry of DNA Damage Induced by Polycyclic Aromatic Hydrocarbons; Relevance for Exposure Monitoring and Risk Assessment

1994 ◽  
Vol 13 (12) ◽  
pp. 880-887 ◽  
Author(s):  
R.A. Baan ◽  
M.-J.S.T. Steenwinkel ◽  
P.T.M. van den Berg ◽  
R. Roggeband ◽  
J.H.M. van Delft
Keyword(s):  
Dna Damage ◽  
Dna Adducts ◽  
Aromatic Hydrocarbons ◽  
Blood Cells ◽  
Adduct Formation ◽  
Dna Adduct ◽  
Immunochemical Analysis ◽  
Basal Cells ◽  
Polycyclic Aromatic

Polycyclic aromatic hydrocarbons (PAH) form a large group of organic chemicals that are widely distributed in our environment as pollutants of air, water and soil. Several PAH are carcinogenic in rodents, while exposure to these compounds has been associated with various types of human cancer. Upon entering the body, PAH may be converted into reactive electrophilic species, which can give rise to the formation of DNA adducts. DNA adduct formation is considered to be the initial event in chemical carcinogenesis. In this paper, two methods are illustrated that are widely used to determine PAH-DNA adduct formation, namely 32P-postlabelling, and immunochemical analysis with specific antibodies. The applications of the 32P-postlabelling assay comprise the following: - A study of interspecies differences in PAH bioactivation in vitro, with microsomal preparations isolated from liver tissue of various rodent species and of human origin; the results indicate that there are considerable qualitative differences between the adduct patterns obtained, which is relevant with respect to extrapolation from animal to man. - The analysis of DNA adduct formation in fish retrieved from marine environments polluted to various extents with PAH; results of these studies show a correlation between liver-DNA adduct levels in these fish and the degree of PAH contamination in the aquatic environment. - Biomonitoring of PAH exposure through analysis of adducts in blood cells obtained from heavy and light smokers; the data show a fair correlation between PAH-DNA adduct levels in white blood cells and cotinine content in blood plasma, the latter being used as a marker for exposure to cigarette smoke. The activity of the detoxifying enzyme glutathione S-transferase M (GSTM1) was also determined in these individuals. Immunochemical analysis with a benzo(a)pyrene(BP)-DNA-specific antiserum was used to investigate BP-adduct induction in situ, in different epithelial cell types—basal/non—basal cells—of hamster trachea exposed to BP in vitro, Histochemical staining of cell-specific cytokeratins was combined with adduct-specific immunostaining. The latter was quantified by immunofluorescence microscopy. The results show that removal of DNA adducts from the basal cells is more rapid during the first 24 h following exposure than from the non-basal cells. The sensitive methods for molecular dosimetry of DNA damage, as illustrated in this paper, appear suitable for determining exposure of animals and humans to PAH. Further animal experiments and in vitro model studies will provide useful additional information that will help evaluate the relevance of biomonitoring data with respect to the health risk that may be associated with the exposure.

scholarly journals Modulation of Carcinogen Metabolism by Nitric Oxide-Aspirin 2 Is Associated with Suppression of DNA Damage and DNA Adduct Formation

2009 ◽  
Vol 284 (33) ◽  
pp. 22099-22107 ◽  
Author(s):  
Christopher J. MacDonald ◽  
Robert Y. S. Cheng ◽  
David D. Roberts ◽  
David A. Wink ◽  
Grace Chao Yeh
Keyword(s):  
Nitric Oxide ◽  
Dna Damage ◽  
Adduct Formation ◽  
Dna Adduct ◽  
Carcinogen Metabolism

DNA damage induced by coal dust, fly and bottom ash from coal combustion evaluated using the micronucleus test and comet assay in vitro

2017 ◽  
Vol 324 ◽  
pp. 781-788 ◽  
Author(s):  
Cristina Araujo Matzenbacher ◽  
Ana Letícia Hilario Garcia ◽  
Marcela Silva dos Santos ◽  
Caroline Cardoso Nicolau ◽  
Suziane Premoli ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Coal Combustion ◽  
Micronucleus Test ◽  
Coal Dust ◽  
Bottom Ash

scholarly journals Corrigendum to: Standardisation of the in vitro comet assay: influence of lysis time and lysis solution composition on the detection of DNA damage induced by X-rays

Mutagenesis ◽  
2019 ◽  
Vol 34 (5-6) ◽  
pp. 431-431
Author(s):  
José M Enciso ◽  
Kristine B Gutzkow ◽  
Gunnar Brunborg ◽  
Ann-Karin Olsen ◽  
Adela López de Cerain ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
X Rays ◽  
Solution Composition ◽  
Lysis Time

scholarly journals Evaluation of Inflammation by Cytokine Production Following Combined Exposure to Ultraviolet and Radiofrequency Radiation of Mobile Phones on 3D Reconstructed Human Skin In Vitro

2020 ◽  
Vol 17 (12) ◽  
pp. 4401
Author(s):  
Zsófia Szilágyi ◽  
Zsuzsanna Németh ◽  
József Bakos ◽  
Péter Pál Necz ◽  
Anna Sáfár ◽  
...  
Keyword(s):  
Human Skin ◽  
Uv Radiation ◽  
Enzyme Concentration ◽  
Protective Effects ◽  
Mobile System ◽  
Wireless Devices ◽  
Rf Exposure ◽  
Solar Uv ◽  
Reconstructed Human Skin

The absorption of exposure to radiofrequency (RF) emitted by wireless devices leads to a high specific absorption rate in the skin. Ultraviolet (UV) radiation can induce several damages to the skin. The aim of this study was to examine whether combined, consecutive exposure to solar UV radiation and 1950 MHz RF exposure of third generation (3G) mobile system have any effect on inflammation processes in the skin. Under in vitro experiments, the inflammation process was examined by cytokines (IL-1α, IL-6, and IL-8) and MMP-1 enzyme secretion on 3D full thickness human skin model. The RF exposure was applied before or after UV irradiation, in order to study either the possible cooperative or protective effects of exposure to RF and UV. We did not find changes in cytokines due to exposure to RF alone. The RF exposure did not enhance the effects of UV radiation. There was a statistically not-significant decrease in cytokines when the skin tissues were pre-exposed to RF before being exposed to 4 standard erythemal dose (SED) UV compared to UV exposure alone. We found that RF exposure reduced the previously UV-treated MMP-1 enzyme concentration. This study might support the evaluation of the effects on the skin exposed to microwave radiation of 5G mobile technology.

DNA adduct formation by secondary metabolites of cyclopenta[cd]pyrene in vitro

1999 ◽  
Vol 136 (2) ◽  
pp. 137-141 ◽  
Author(s):  
Ching-Hung Hsu ◽  
Paul L. Skipper ◽  
Steven R. Tannenbaum
Keyword(s):  
Secondary Metabolites ◽  
Adduct Formation ◽  
Dna Adduct

scholarly journals In vitro evaluation of baseline and induced DNA damage in human sperm exposed to benzo[a]pyrene or its metabolite benzo[a]pyrene-7,8-diol-9,10-epoxide, using the comet assay

Mutagenesis ◽  
2010 ◽  
Vol 25 (4) ◽  
pp. 417-425 ◽  
Author(s):  
V. Sipinen ◽  
J. Laubenthal ◽  
A. Baumgartner ◽  
E. Cemeli ◽  
J. O. Linschooten ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Human Sperm ◽  
In Vitro Evaluation
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