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Medical Hypotheses Jun 2019DNA adducts are associated with a number of diseases, including cancer. Based on a recent report by our group, the aim of this study was to test the hypothesis that DNA...
DNA adducts are associated with a number of diseases, including cancer. Based on a recent report by our group, the aim of this study was to test the hypothesis that DNA adducts can be removed by means of one or more of the following three intervention programmes: intermittent whole-body hyperthermia; detoxification; and cell repair. The number of DNA adducts and total DNA adduct concentrations were measured in 104 patients who underwent one or more of the three intervention programmes. DNA adduct assessments were carried out on extracted genomic DNA by gas-liquid chromatography, with any DNA adducts found being localised using DNA microarrays. The baseline median number of DNA adducts was 2. The follow-up median number of adducts was highly significantly lower at 0 (p < 0.000000000000003). The mean total DNA adduct concentration at baseline was 9.308 ng/mL, and highly significantly lower at follow-up at 1.553 ng/mL (p < 0.000000000000006). Of the three intervention programmes, only the intermittent whole-body hyperthermia was associated with a significant reduction in DNA adducts. This study offers support for the hypothesis that DNA adducts can be removed by intermittent whole-body hyperthermia. The intermittent hyperthermia used involved infrared-A (wavelength 700-1400 nm, or, equivalently, a frequency of 215-430 THz) being preferentially delivered to the whole body, apart from the head, for up to one hour per session, with gradual core body temperature elevation usually occurring during the first 20-30 min. These results may offer an explanation at the molecular level for other reported clinical benefits of intermittent whole-body hyperthermia.
Topics: Administration, Intravenous; Chromatography, Gas; DNA Adducts; Fatty Acids; Female; Humans; Hyperthermia, Induced; Male; Middle Aged; Models, Biological; Neoplasms; Oligonucleotide Array Sequence Analysis; Parkinson Disease; Phospholipids; Reproducibility of Results
PubMed: 31088632
DOI: 10.1016/j.mehy.2019.03.031 -
Methods in Molecular Biology (Clifton,... 2020P-Postlabeling analysis is an ultra-sensitive method for the detection of DNA adducts, such as those formed directly by the covalent binding of carcinogens and mutagens...
P-Postlabeling analysis is an ultra-sensitive method for the detection of DNA adducts, such as those formed directly by the covalent binding of carcinogens and mutagens to bases in DNA, and other DNA lesions resulting from modification of bases by endogenous or exogenous agents (e.g., oxidative damage). The procedure involves four main steps: enzymatic digestion of DNA sample; enrichment of the adducts; radiolabeling of the adducts by T4 kinase-catalyzed transference of P-orthophosphate from [γ-P]ATP; chromatographic separation of labeled adducts, and detection and quantification by means of their radioactive decay. Using 10 μg of DNA or less, it is capable of detecting adduct levels as low as 1 adduct in 10-10 normal nucleotides. It is applicable to a wide range of investigations, including monitoring human exposure to environmental or occupational carcinogens, determining whether a chemical has genotoxic properties, analysis of the genotoxicity of complex mixtures, elucidation of the pathways of activation of carcinogens, and monitoring DNA repair.
Topics: Animals; Carcinogens; Chromatography, High Pressure Liquid; DNA Adducts; DNA Damage; Fungal Proteins; Humans; Isotope Labeling; Mutagens; Oxidative Stress; Phosphorus Radioisotopes; Phosphotransferases; Single-Strand Specific DNA and RNA Endonucleases; Workflow
PubMed: 31989562
DOI: 10.1007/978-1-0716-0223-2_16 -
The International Journal of... Sep 2023Cisplatin, a widely prescribed chemotherapeutic agent for treating solid tumors, induces DNA adducts and activates cellular defense mechanisms, including DNA repair,...
Cisplatin, a widely prescribed chemotherapeutic agent for treating solid tumors, induces DNA adducts and activates cellular defense mechanisms, including DNA repair, cell cycle checkpoint control, and apoptosis. Considering the circadian rhythmicity displayed by most chemotherapeutic agents and their varying therapeutic efficacy based on treatment timing, our study aimed to investigate whether the circadian clock system influences the DNA damage responses triggered by cisplatin in synchronized cells. We examined the DNA damage responses in circadian-synchronized wild-type mouse embryonic fibroblasts (WT-MEF; clock-proficient cells), cryptochrome1 and 2 double knock-out MEF (CRY; clock-deficient cells), and mouse hepatocarcinoma Hepa1c1c7 cells. Varying the treatment time resulted in a significant difference in the rate of platinum-DNA adduct removal specifically in circadian-synchronized WT-MEF, while CRY did not exhibit such variation. Moreover, diurnal variation in other DNA damage responses, such as cell cycle checkpoint activity indicated by p53 phosphorylation status and apoptosis measured by DNA break frequency, was observed only in circadian-synchronized WT-MEF, not in CRY or mouse hepatocarcinoma Hepa1c1c7 cells. These findings highlight that the DNA damage responses triggered by cisplatin are indeed governed by circadian control exclusively in clock-proficient cells. This outcome bears potential implications for enhancing or devising chronotherapy approaches for cancer patients.
Topics: Animals; Mice; Cisplatin; DNA Adducts; DNA Damage; Fibroblasts; DNA Repair; Circadian Clocks; Neoplasms; Apoptosis
PubMed: 37574041
DOI: 10.1016/j.biocel.2023.106454 -
Journal of Applied Genetics Aug 2017Investigations on the impact of chemicals on the environment and human health have led to the development of an exposome concept. The exposome refers to the totality of... (Review)
Review
Investigations on the impact of chemicals on the environment and human health have led to the development of an exposome concept. The exposome refers to the totality of exposures received by a person during life, including exposures to life-style factors, from the prenatal period to death. The exposure to genotoxic chemicals and their reactive metabolites can induce chemical modifications of DNA, such as, for example, DNA adducts, which have been extensively studied and which play a key role in chemically induced carcinogenesis. Development of different methods for the identification of DNA adducts has led to adopting DNA adductomic approaches. The ability to simultaneously detect multiple PAH-derived DNA adducts may allow for the improved assessment of exposure, and offer a mechanistic insight into the carcinogenic process following exposure to PAH mixtures. The major advantage of measuring chemical-specific DNA adducts is the assessment of a biologically effective dose. This review provides information about the occurrence of the polycyclic aromatic hydrocarbons (PAHs) and their influence on human exposure and biological effects, including PAH-derived DNA adduct formation and repair processes. Selected methods used for determination of DNA adducts have been presented.
Topics: Biotransformation; DNA Adducts; DNA Damage; DNA Repair; Environmental Exposure; Humans; Polycyclic Aromatic Hydrocarbons
PubMed: 27943120
DOI: 10.1007/s13353-016-0380-3 -
DNA Repair Oct 2020Proteins that act on DNA, or are in close proximity to it, can become inadvertently crosslinked to DNA and form highly toxic lesions, known as DNA-protein crosslinks...
Proteins that act on DNA, or are in close proximity to it, can become inadvertently crosslinked to DNA and form highly toxic lesions, known as DNA-protein crosslinks (DPCs). DPCs are generated by different chemotherapeutics, environmental or endogenous sources of crosslinking agents, or by lesions on DNA that stall the catalytic cycle of certain DNA processing enzymes. These bulky adducts impair processes on DNA such as DNA replication or transcription, and therefore pose a serious threat to genome integrity. The large diversity of DPCs suggests that there is more than one canonical mechanism to repair them. Indeed, many different enzymes have been shown to act on DPCs by either processing the protein, the DNA or the crosslink itself. In addition, the cell cycle stage or cell type are likely to dictate pathway choice. In recent years, a detailed understanding of DPC repair during S phase has started to emerge. Here, we review the current knowledge on the mechanisms of replication-coupled DPC repair, and describe and also speculate on possible pathways that remove DPCs outside of S phase. Moreover, we highlight a recent paradigm shifting finding that indicates that DPCs are not always detrimental, but can also play a protective role, preserving the genome from more deleterious forms of DNA damage.
Topics: Animals; DNA; DNA Adducts; DNA Repair; DNA Replication; Eukaryota; Humans; Proteins
PubMed: 32683310
DOI: 10.1016/j.dnarep.2020.102924 -
Rapid Communications in Mass... Jul 2021As a new approach to DNA adductomics, we directly reacted intact, double-stranded (ds)-DNA under warm conditions with an alkylating mass tag followed by analysis by...
RATIONALE
As a new approach to DNA adductomics, we directly reacted intact, double-stranded (ds)-DNA under warm conditions with an alkylating mass tag followed by analysis by liquid chromatography/mass spectrometry. This method is based on the tendency of adducted nucleobases to locally disrupt the DNA structure (forming a "DNA bubble") potentially increasing exposure of their nucleophilic (including active hydrogen) sites for preferential alkylation. Also encouraging this strategy is that the scope of nucleotide excision repair is very broad, and this system primarily recognizes DNA bubbles.
METHODS
A cationic xylyl (CAX) mass tag with limited nonpolarity was selected to increase the retention of polar adducts in reversed-phase high-performance liquid chromatography (HPLC) for more detectability while maintaining resolution. We thereby detected a diversity of DNA adducts (mostly polar) by the following sequence of steps: (1) react DNA at 45°C for 2 h under aqueous conditions with CAX-B (has a benzyl bromide functional group to label active hydrogen sites) in the presence of triethylamine; (2) remove residual reagents by precipitating and washing the DNA (a convenient step); (3) digest the DNA enzymatically to nucleotides and remove unlabeled nucleotides by nonpolar solid-phase extraction (also a convenient step); and (4) detect CAX-labeled, adducted nucleotides by LC/MS or a matrix-assisted laser desorption/ionization (MALDI)-MS technique.
RESULTS
Examples of the 42 DNA or RNA adducts detected, or tentatively so based on accurate mass and fragmentation data, are as follows: 8-oxo-dGMP, ethyl-dGMP, hydroxyethyl-dGMP (four isomers, all HPLC-resolved), uracil-glycol, apurinic/apyrimidinic sites, benzo[a]pyrene-dGMP, and, for the first time, benzoquinone-hydroxymethyl-dCMP. Importantly, these adducts are detected in a single procedure under a single set of conditions. Sensitivity, however, is only defined in a preliminary way, namely the latter adduct seems to be detected at a level of about 4 adducts in 10 nucleotides (S/N ~30).
CONCLUSIONS
CAX-Prelabeling is an emerging new technique for DNA adductomics, providing polar DNA adductomics in a practical way for the first time. Further study of the method is encouraged to better characterize and extend its performance, especially in scope and sensitivity.
Topics: Animals; Benzo(a)pyrene; Benzyl Compounds; Cations; Cattle; Chromatography, High Pressure Liquid; DNA Adducts; Ethylamines; Guanine; Humans; Nucleotides; Phosphorus Radioisotopes; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Uracil
PubMed: 33821547
DOI: 10.1002/rcm.9095 -
Mutation Research. Genetic Toxicology... 2021In 2015, the International Agency for Research on Cancer classified the consumption of processed meat as carcinogenic to humans (Group 1) and red meat as probably... (Review)
Review
In 2015, the International Agency for Research on Cancer classified the consumption of processed meat as carcinogenic to humans (Group 1) and red meat as probably carcinogenic to humans (Group 2A) based on sufficient data from animal models and epidemiological studies. However, research characterising the mechanisms underlying this carcinogenic process in humans are limited, particularly with respect to measures of direct DNA damage. The current review sought to evaluate and summarize the recent literature, published since 2000, regarding the associations of meat consumption and three biomarkers of genotoxicity in humans: DNA strand breaks (measured using the comet assay), DNA adducts, and micronucleus formation. After screening 230 potential articles, 35 were included, and then were classified as experimental or observational in design, the latter of which were further categorized according to their dietary assessment approach. Among the 30 observational studies, 4 of which used two different assays, 3 of 5 comet assay studies, 13 of 20 DNA adduct studies, and 7 of 9 micronucleus studies reported a positive association between meat consumption and DNA damage. Among the 5 experimental studies, 1 of 1 using the comet assay, 3 of 3 measuring DNA adducts and 0 of 1 measuring micronuclei reported significant positive associations with meat consumption. Nevertheless, common limitations among the selected publications included small sample size, and poor methodological reporting of both exposure and outcome measures. Moreover, the vast majority of studies only measured DNA damage in one biological sample using a single assay and we cannot exclude the possibility of publication bias. Ultimately, our review of the literature, published since 2000, revealed a preponderance of studies that support mechanisms of genotoxicity in playing an important role in the meat-cancer association.
Topics: Comet Assay; DNA Adducts; DNA Damage; Food Preferences; Humans; Micronucleus Tests; Red Meat
PubMed: 33678247
DOI: 10.1016/j.mrgentox.2021.503311 -
Environmental and Molecular Mutagenesis Aug 2016Over two centuries ago, Sir Percival Pott, a London surgeon, published a pioneering treatise showing that soot exposure was the cause of high incidences of scrotal... (Review)
Review
Over two centuries ago, Sir Percival Pott, a London surgeon, published a pioneering treatise showing that soot exposure was the cause of high incidences of scrotal cancers occurring in young men who worked as chimney sweeps. Practicing at a time when cellular pathology was not yet recognized, Sir Percival nonetheless observed that the high incidence and short latency of the chimney sweep cancers, was fundamentally different from the rare scrotal cancers typically found in elderly men. Furthermore, his diagnosis that the etiology of these cancers was related to chimney soot exposure, was absolutely accurate, conceptually novel, and initiated the field of "occupational cancer epidemiology." After many intervening years of research focused on mechanisms of chemical carcinogenesis, briefly described here, it is clear that DNA damage, or DNA adduct formation, is "necessary but not sufficient" for tumor induction, and that many additional factors contribute to carcinogenesis. This review includes a synopsis of carcinogen-induced DNA adduct formation in experimental models and in the human population, with particular attention paid to molecular dosimetry and molecular cancer epidemiology. Environ. Mol. Mutagen. 57:499-507, 2016. © 2016 Wiley Periodicals, Inc.
Topics: Animals; Carcinogenesis; Carcinogens, Environmental; DNA Adducts; Environmental Exposure; Humans; Molecular Epidemiology; Neoplasms
PubMed: 27346877
DOI: 10.1002/em.22030 -
Environmental Toxicology and... Jun 2024The concept of the exposome is the encompassing of all the environmental exposures, both exogenous and endogenous, across the life course. Many, if not all, of these... (Review)
Review
The concept of the exposome is the encompassing of all the environmental exposures, both exogenous and endogenous, across the life course. Many, if not all, of these exposures can result in the generation of reactive species, and/or the modulation of cellular processes, that can lead to a breadth of modifications of DNA, the nature of which may be used to infer their origin. Because of their role in cell function, such modifications have been associated with various major human diseases, including cancer, and so their assessment is crucial. Historically, most methods have been able to only measure one or a few DNA modifications at a time, limiting the information available. With the development of DNA adductomics, which aims to determine the totality of DNA modifications, a far more comprehensive picture of the DNA adduct burden can be gained. Importantly, DNA adductomics can facilitate a "top-down" investigative approach whereby patterns of adducts may be used to trace and identify the originating exposure source. This, together with other 'omic approaches, represents a major tool for unraveling the complexities of the exposome and hence allow a better a understanding of the environmental origins of disease.
Topics: Humans; DNA Adducts; Exposome; Environmental Exposure; Biomarkers; Animals; DNA
PubMed: 38636743
DOI: 10.1016/j.etap.2024.104449 -
Journal of Immunoassay & Immunochemistry 2018Arginine is derived from dietary intake, body protein breakdown, or endogenous de novo arginine production. Arginine methylation of non-histone proteins is used in... (Review)
Review
Arginine is derived from dietary intake, body protein breakdown, or endogenous de novo arginine production. Arginine methylation of non-histone proteins is used in transcriptional regulation. Protein-arginine methylation is used for regulation of transcriptional and various physiological pathological processes. Protein methylation may affect protein-protein, protein-DNA, or protein-RNA interaction. Arginine has an effect on the DNA-binding activity of NF-κB, a dominant transcriptional factor in inflammation. Adduct formation results in increased secretion of messenger molecules such as cytokines and chemokines that mediate communication among cells and promote inflammation. Arginine and lysine amino acid-rich histones in nucleosomes on modification by environmental agents form histone-DNA adducts, making it immunogenic. Alteration of DNA resulting from photomodification could lead to the development of antibodies or mutations to modified DNA. Lysine and arginine-rich histones in nucleosomes on modification by environmental agents form histone-DNA adducts, making it immunogenic. Alteration of DNA resulting from photomodification could lead to the development of antibodies or mutations to modified DNA. Therefore, the DNA-arginine photoadduct and modified photoadduct could have important implications in various pathophysiological conditions such as toxicology, carcinogenesis, and autoimmune phenomena. Abbreviations: Arg: Arginine; SLE: systemic lupus erythematosus; UV: ultraviolet; Tm: thermal melting temperature; NO: nitric oxide; O.: superoxide anion.
Topics: Animals; Arginine; Autoantibodies; DNA Adducts; Humans
PubMed: 30362913
DOI: 10.1080/15321819.2018.1536667