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Aristolochic acids I and II (AA-I/II) are carcinogenic principles of Aristolochia plants, which have been employed in traditional medicinal practices and discovered as food contaminants. While the deleterious effects of AAs are broadly acknowledged, there is a dearth of information to define the mechanisms underlying their carcinogenicity. Following bioactivation in the liver, N-hydroxyaristolactam and N-sulfonyloxyaristolactam metabolites are transported via circulation and elicit carcinogenic effects by reacting with cellular DNA. In this study, we apply DNA adduct analysis, X-ray crystallography, isothermal titration calorimetry, and fluorescence quenching to investigate the role of human serum albumin (HSA) in modulating AA carcinogenicity. We find that HSA extends the half-life and reactivity of N-sulfonyloxyaristolactam-I with DNA, thereby protecting activated AAs from heterolysis. Applying novel pooled plasma HSA crystallization methods, we report high-resolution structures of myristic acid-enriched HSA (HSAMYR) and its AA complexes (HSAMYR/AA-I and HSAMYR/AA-II) at 1.9 Å resolution. While AA-I is located within HSA subdomain IB, AA-II occupies subdomains IIA and IB. ITC binding profiles reveal two distinct AA sites in both complexes with association constants of 1.5 and 0.5 · 106 M-1 for HSA/AA-I versus 8.4 and 9.0 · 105 M-1 for HSA/AA-II. Fluorescence quenching of the HSA Trp214 suggests variable impacts of fatty acids on ligand binding affinities. Collectively, our structural and thermodynamic characterizations yield significant insights into AA binding, transport, toxicity, and potential allostery, critical determinants for elucidating the mechanistic roles of HSA in modulating AA carcinogenicity.
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Ácidos Aristolóquicos , Albumina Sérica Humana , Ácidos Aristolóquicos/metabolismo , Ácidos Aristolóquicos/química , Humanos , Cristalografia por Raios X , Albumina Sérica Humana/metabolismo , Albumina Sérica Humana/química , Adutos de DNA/metabolismo , Adutos de DNA/química , Ligação Proteica , Ácido Mirístico/metabolismo , Ácido Mirístico/químicaRESUMO
PURPOSE: In East Asia, the incidence of breast cancer has been increasing rapidly, particularly among premenopausal women. An elevated ratio of estrogen-DNA adducts was linked to a higher risk of breast cancer. The present study explored the influence of the interaction between base excision repair (BER) gene polymorphisms and estrogen-DNA adducts on breast cancer risk. METHODS: We conducted a case-control study comprising healthy volunteers and individuals with benign breast disease (control arm, n = 176) and patients with invasive carcinoma or carcinoma in situ (case arm, n = 177). Genotyping for BER-related genes, including SMUG1, OGG1, ERCC5, and APEX1, was performed. A logistic regression model, incorporating interactions between gene polymorphisms, estrogen-DNA adduct ratio, and clinical variables, was used to identify the risk factors for breast cancer. RESULTS: Univariate analysis indicated marginal associations between breast cancer risk and APEX1 rs1130409 T > G (P = 0.057) and APEX1 rs1760944 T > G (P = 0.065). Multivariate regression analysis revealed significant associations with increased breast cancer risk for APEX1_rs1130409 (GT/GG versus TT) combined with a natural logarithmic value of the estrogen-DNA adduct ratio (estimated OR 1.164, P = 0.023) and premenopausal status with an estrogen-DNA adduct ratio > 2.93 (estimated OR 2.433, P = 0.001). CONCLUSION: APEX1_rs1130409 (GT/GG versus TT) polymorphisms, which are related to decreased BER activity, combined with an increased ratio of estrogen-DNA adducts, increase the risk of breast cancer in East Asian women.
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CONTEXT: Exocyclic DNA adducts have been shown to be potential biomarkers of cancer risk related to oxidative stress and exposure to aldehydes in smokers. In fact, aldehydes potentially arise from tobacco combustion directly and endogenously through lipid peroxidation. OBJECTIVE: This study aims to investigate the relationship between a profile of nine aldehydes-induced DNA adducts and antioxidant activities, in order to evaluate new biomarkers of systemic exposure to aldehydes. METHODS: Using our previously published UPLC-MS/MS method, adducts levels were quantified in the blood DNA of 34 active smokers. The levels of antioxidant vitamins (A, C and E), coenzyme Q10, ß-carotene, superoxide dismutase (SOD) and autoantibodies against oxidized low-density lipoprotein were measured. RESULTS: Adducts induced by tobacco smoking-related aldehydes were quantified at levels reflecting an oxidative production from lipid peroxidation. A significant correlation between SOD and crotonaldehyde-induced adducts (p = 0.0251) was also observed. ß-Carotene was negatively correlated with the adducts of formaldehyde (p = 0.0351) and acetaldehyde (p = 0.0413). Vitamin C tended to inversely correlate with acetaldehyde-induced adducts (p = 0.0584). CONCLUSION: These results are promising, and the study is now being conducted on a larger cohort with the aim of evaluating the impact of smoking cessation programs on the evolution of adducts profile and antioxidants activities.
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Adutos de DNA , Fumantes , Humanos , Monitoramento Biológico , Antioxidantes , beta Caroteno , Cromatografia Líquida , Espectrometria de Massas em Tandem , Aldeídos , Estresse Oxidativo , Biomarcadores , Acetaldeído , Superóxido DismutaseRESUMO
Large interspecies differences between rats and mice concerning the hepatotoxicity and carcinogenicity of aflatoxin B1 (AFB1) are known, with mice being more resistant. However, a comprehensive interspecies comparison including subcellular liver tissue compartments has not yet been performed. In this study, we performed spatio-temporal intravital analysis of AFB1 kinetics in the livers of anesthetized mice and rats. This was supported by time-dependent analysis of the parent compound as well as metabolites and adducts in blood, urine, and bile of both species by HPLC-MS/MS. The integrated data from intravital imaging and HPLC-MS/MS analysis revealed major interspecies differences between rats and mice: (1) AFB1-associated fluorescence persisted much longer in the nuclei of rat than mouse hepatocytes; (2) in the sinusoidal blood, AFB1-associated fluorescence was rapidly cleared in mice, while a time-dependent increase was observed in rats in the first three hours after injection followed by a plateau that lasted until the end of the observation period of six hours; (3) this coincided with a far stronger increase of AFB1-lysine adducts in the blood of rats compared to mice; (4) the AFB1-guanine adduct was detected at much higher concentrations in bile and urine of rats than mice. In both species, the AFB1-glutathione conjugate was efficiently excreted via bile, where it reached concentrations at least three orders of magnitude higher compared to blood. In conclusion, major differences between mice and rats were observed, concerning the nuclear persistence, formation of AFB1-lysine adducts, and the AFB1-guanine adducts.
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Aflatoxinas , Ratos , Camundongos , Animais , Aflatoxinas/metabolismo , Aflatoxinas/toxicidade , Lisina/metabolismo , Espectrometria de Massa com Cromatografia Líquida , Espectrometria de Massas em Tandem , Fígado/metabolismo , Aflatoxina B1/toxicidade , Guanina/metabolismo , Microscopia IntravitalRESUMO
We previously found that feeding rats with broccoli or cauliflower leads to the formation of characteristic DNA adducts in the liver, intestine and various other tissues. We identified the critical substances in the plants as 1-methoxy-3-indolylmethyl (1-MIM) glucosinolate and its degradation product 1-MIM-OH. DNA adduct formation and the mutagenicity of 1-MIM-OH in cell models were drastically enhanced when human sulfotransferase (SULT) 1A1 was expressed. The aim of this study was to clarify the role of SULT1A1 in DNA adduct formation by 1-MIM-OH in mouse tissues in vivo. Furthermore, we compared the endogenous mouse Sult1a1 and transgenic human SULT1A1 in the activation of 1-MIM-OH using genetically modified mouse strains. We orally treated male wild-type (wt) and Sult1a1-knockout (ko) mice, as well as corresponding lines carrying the human SULT1A1-SULT1A2 gene cluster (tg and ko-tg), with 1-MIM-OH. N2-(1-MIM)-dG and N6-(1-MIM)-dA adducts in DNA were analysed using isotope-dilution UPLC-MS/MS. In the liver, caecum and colon adducts were abundant in mice expressing mouse and/or human SULT1A1, but were drastically reduced in ko mice (1.2-10.6% of wt). In the kidney and small intestine, adduct levels were high in mice carrying human SULT1A1-SULT1A2 genes, but low in wt and ko mice (1.8-6.3% of tg-ko). In bone marrow, adduct levels were very low, independently of the SULT1A1 status. In the stomach, they were high in all four lines. Thus, adduct formation was primarily controlled by SULT1A1 in five out of seven tissues studied, with a strong impact of differences in the tissue distribution of mouse and human SULT1A1. The behaviour of 1-MIM-OH in these models (levels and tissue distribution of DNA adducts; impact of SULTs) was similar to that of methyleugenol, classified as "probably carcinogenic to humans". Thus, there is a need to test 1-MIM-OH for carcinogenicity in animal models and to study its adduct formation in humans consuming brassicaceous foodstuff.
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Adutos de DNA , Glucosinolatos , Camundongos , Humanos , Animais , Ratos , Camundongos Knockout , Cromatografia Líquida , Espectrometria de Massas em Tandem , Arilsulfotransferase/genéticaRESUMO
It is generally accepted that adjacent guanine residues in DNA are the primary target for platinum antitumor drugs and that differences in the conformations of the Pt-DNA adducts can play a role in their antitumor activity. In this study, we investigated the effect of the carrier ligand cis-1,3-diaminocyclohexane (cis-1,3-DACH) upon formation, stability, and stereochemistry of the (cis-1,3-DACH)PtG2 and (cis-1,3-DACH)Pt(d(GpG)) adducts (G = 9-EthlyGuanine, guanosine, 5'- and 3'-guanosine monophosphate; d(GpG) = deoxyguanosil(3'-5')deoxyguanosine). A peculiar feature of the cis-1,3-DACH carrier ligand is the steric bulk of the diamine, which is asymmetric with respect to the Pt-coordination plane. The (cis-1,3-DACH)Pt(5'GMP)2 and (cis-1,3-DACH)Pt(3'GMP)2 adducts show preference for the ΛHT and ∆HT conformations, respectively (HT stands for Head-to-Tail). Moreover, the increased intensity of the circular dichroism signals in the cis-1,3-DACH derivatives with respect to the analogous cis-(NH3)2 species could be a consequence of the greater bite angle of the cis-1,3-DACH carrier ligand with respect to cis-(NH3)2. Finally, the (cis-1,3-DACH)Pt(d(GpG)) adduct is present in two isomeric forms, each one giving a pair of H8 resonances linked by a NOE cross peak. The two isomers were formed in comparable amounts and had a dominance of the HH conformer but with some contribution of the ΔHT conformer which is related to the HH conformer by having the 3'-G base flipped with respect to the 5'-G residue.
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Adutos de DNA , DNA , Oxaliplatina , DNA/química , DNA/metabolismo , Adutos de DNA/química , Oxaliplatina/química , Oxaliplatina/farmacologia , Compostos Organoplatínicos/química , Compostos Organoplatínicos/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Ligantes , Modelos Moleculares , Conformação de Ácido NucleicoRESUMO
Lung cancer is the leading cause of cancer death worldwide. Polycyclic aromatic hydrocarbons (PAHs) are metabolized by the cytochrome P450 (CYP)1A and 1B1 to DNA-reactive metabolites, which could lead to mutations in critical genes, eventually resulting in cancer. Omega-3 fatty acids, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are beneficial against cancers. In this investigation, we elucidated the mechanisms by which omega-3 fatty acids EPA and DHA will attenuate PAH-DNA adducts and lung carcinogenesis and tumorigenesis mediated by the PAHs BP and MC. Adult wild-type (WT) (A/J) mice, Cyp1a1-null, Cyp1a2-null, or Cyp1b1-null mice were exposed to PAHs benzo[a]pyrene (BP) or 3-methylcholanthrene (MC), and the effects of omega-3 fatty acid on PAH-mediated lung carcinogenesis and tumorigenesis were studied. The major findings were as follows: (i) omega-3 fatty acids significantly decreased PAH-DNA adducts in the lungs of each of the genotypes studied; (ii) decreases in PAH-DNA adduct levels by EPA/DHA was in part due to inhibition of CYP1B1; (iii) inhibition of soluble epoxide hydrolase (sEH) enhanced the EPA/DHA-mediated prevention of pulmonary carcinogenesis; and (iv) EPA/DHA attenuated PAH-mediated carcinogenesis in part by epigenetic mechanisms. Taken together, our results suggest that omega-3 fatty acids have the potential to be developed as cancer chemo-preventive agents in people.
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Ácidos Graxos Ômega-3 , Hidrocarbonetos Policíclicos Aromáticos , Humanos , Adulto , Camundongos , Animais , Ácidos Graxos Ômega-3/farmacologia , Adutos de DNA , Carcinogênese , Transformação Celular Neoplásica , Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/farmacologiaRESUMO
The SARS-CoV-2 helicase, non-structural protein 13 (Nsp13), plays an essential role in viral replication, translocating in the 5' â 3' direction as it unwinds double-stranded RNA/DNA. We investigated the impact of structurally distinct DNA lesions on DNA unwinding catalyzed by Nsp13. The selected lesions include two benzo[a]pyrene (B[a]P)-derived dG adducts, the UV-induced cyclobutane pyrimidine dimer (CPD), and the pyrimidine (6-4) pyrimidone (6-4PP) photolesion. The experimentally observed unwinding rate constants (kobs) and processivities (P) were examined. Relative to undamaged DNA, the kobs values were diminished by factors of up to ~15 for B[a]P adducts but only by factors of ~2-5 for photolesions. A minor-groove-oriented B[a]P adduct showed the smallest impact on P, which decreased by ~11% compared to unmodified DNA, while an intercalated one reduced P by ~67%. However, the photolesions showed a greater impact on the processivities; notably, the CPD, with the highest kobs value, exhibited the lowest P, which was reduced by ~90%. Our findings thus show that DNA unwinding efficiencies are lesion-dependent and most strongly inhibited by the CPD, leading to the conclusion that processivity is a better measure of DNA lesions' inhibitory effects than unwinding rate constants.
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DNA Helicases , SARS-CoV-2 , Proteínas não Estruturais Virais , SARS-CoV-2/metabolismo , Proteínas não Estruturais Virais/metabolismo , Proteínas não Estruturais Virais/química , DNA Helicases/metabolismo , DNA Helicases/química , DNA/metabolismo , DNA/química , Humanos , Dano ao DNA , COVID-19/virologia , Cinética , Metiltransferases , RNA HelicasesRESUMO
DNA adduction in the model yeast Saccharomyces cerevisiae was investigated after exposure to the fungicide penconazole and the reference genotoxic compound benzo(a)pyrene, for validating yeasts as a tool for molecular toxicity studies, particularly of environmental pollution. The effect of the toxicants on the yeast's growth kinetics was determined as an indicator of cytotoxicity. Fermentative cultures of S. cerevisiae were exposed to 2 ppm of Penconazole during different phases of growth; while 0.2 and 2 ppm of benzo(a)pyrene were applied to the culture medium before inoculation and on exponential cultures. Exponential respiratory cultures were also exposed to 0.2 ppm of B(a)P for comparison of both metabolisms. Penconazole induced DNA adducts formation in the exponential phase test; DNA adducts showed a peak of 54.93 adducts/109 nucleotides. Benzo(a)pyrene induced the formation of DNA adducts in all the tests carried out; the highest amount of 46.7 adducts/109 nucleotides was obtained in the fermentative cultures after the exponential phase exposure to 0.2 ppm; whereas in the respiratory cultures, 14.6 adducts/109 nucleotides were detected. No cytotoxicity was obtained in any experiment. Our study showed that yeast could be used to analyse DNA adducts as biomarkers of exposure to environmental toxicants.
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Benzo(a)pireno , Adutos de DNA , Poluentes Ambientais , Saccharomyces cerevisiae , Adutos de DNA/metabolismo , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento , Benzo(a)pireno/toxicidade , Benzo(a)pireno/metabolismo , Poluentes Ambientais/toxicidade , Poluentes Ambientais/metabolismo , Mutagênicos/toxicidade , Mutagênicos/metabolismo , DNA Fúngico/genética , Fungicidas Industriais/toxicidade , Fungicidas Industriais/metabolismoRESUMO
Helix-distorting DNA damages block RNA and DNA polymerase, compromising cell function and fate. In human cells, these damages are removed primarily by nucleotide excision repair (NER). Here, we describe damage-sensing PCR (dsPCR), a PCR-based method for the detection of these DNA damages. Exposure to DNA damaging agents results in lower PCR signal in comparison to non-damaged DNA, and repair is measured as the restoration of PCR signal over time. We show that the method successfully detects damages induced by ultraviolet (UV) radiation, by the carcinogenic component of cigarette smoke benzo[a]pyrene diol epoxide (BPDE) and by the chemotherapeutic drug cisplatin. Damage removal measured by dsPCR in a heterochromatic region is less efficient than in a transcribed and accessible region. Furthermore, lower repair is measured in repair-deficient knock-out cells. This straight-forward method could be applied by non-DNA repair experts to study the involvement of their gene-of-interest in repair. Furthermore, this method is fully amenable for high-throughput screening of DNA repair activity.
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Adutos de DNA , Dano ao DNA , Reparo do DNA , Humanos , Carcinógenos/toxicidade , DNA/efeitos dos fármacos , DNA/efeitos da radiação , Adutos de DNA/análise , Reparo do DNA/genética , Reação em Cadeia da Polimerase/métodosRESUMO
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.
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Biomarcadores , Adutos de DNA , Exposição Ambiental , Expossoma , Humanos , Animais , DNARESUMO
Colorectal cancer (CRC) is the third leading cause of cancer-related mortalities in the USA and around 52,550 people were expected to die from this disease by December 2023. The objective of this study was to investigate the effect of diet type on benzo(a)pyrene [B(a)P]-induced colon cancer in an adult male rat model, the Polyposis In the Rat Colon (PIRC) kindred type. Groups of PIRC rats (n = 10) were fed with AIN-76A regular diet (RD) or Western diet (WD) and received 25, 50 and 100 µg B(a)P/kg body wt. via oral gavage for 60 days. Rats fed diets alone, but no B(a)P, served as controls. After exposure, rats were euthanized; colon and liver samples were analyzed for activation of drug metabolizing enzymes (DMEs) CYP1A1, CYP1B1, SULT and GST. Plasma and tissue samples were analyzed by reverse phase-HPLC for B(a)P metabolites. In addition to these studies, DNA isolated from colon and liver tissues was analyzed for B(a)P-induced DNA adducts by the 32P-postlabeling method using a thin-layer chromatography system. Western diet consumption resulted in a marked increase in DME expression and B(a)P metabolite concentrations in rats that were administered 100 µg/kg B(a)P + WD (p < 0.05) compared to other treatment groups. Our findings demonstrate that WD accelerates the development of colon tumors induced by B(a)P through enhanced biotransformation, and the products of this process (metabolites) were found to bind with DNA and form B(a)P-DNA adducts, which may have given rise to colon polyps characterized by gain in tumor number, sizes, and dysplasia.
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Urbanization has numerous benefits to human society, but some aspects of urban environments, such as air pollution, can negatively affect human health. Two major air pollutants, particulate matter (PM) and polycyclic aromatic hydrocarbons (PAH), have been classified as carcinogens by the International Agency for Research on Cancer. Here, we answer two questions: (1) What are the carcinogenic effects of PM and PAH exposure? (2) How does carcinogenic risk vary across geographical regions? We performed a comprehensive literature search of peer-reviewed published studies examining the link between air pollution and human cancer rates. Focusing on studies published since 2014 when the last IARC monograph on air pollution was published, we converted the extracted data into relative risks and performed subgroup analyses. Exposure to PM2.5 (per 10 µg/m3) resulted in an 8.5% increase in cancer incidence when all cancer types were combined, and risk for individual cancer types (i.e. lung cancer and adenocarcinoma) was also elevated. PM2.5 was also associated with 2.5% higher mortality due to cancer when all types of cancer were combined, and for individual cancer types (i.e., lung and breast cancer). Exposure to PM2.5 and PM10 posed the greatest risk to lung cancer incidence and mortality in Europe (PM2.5 RR 2.15; PM10 RR 1.26); the risk in Asia and the Americas was also elevated. Exposure to PAH and benzo[a]pyrene significantly increased the pooled risk of cancer incidence (10.8% and 8.0% respectively) at the highest percentile of exposure concentration. Our meta-analyses of studies over the past decade shows that urban air pollution in the form of PM2.5, PM10, and PAH all elevate the incidence and mortality of cancer. We discuss the possible mechanisms of carcinogenesis of PM and PAH. These results support World Health Organization's conclusion that air pollution poses among the greatest health risks to humans living in cities.
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Poluentes Atmosféricos , Carcinógenos , Neoplasias , Material Particulado , Hidrocarbonetos Policíclicos Aromáticos , Humanos , Poluição do Ar/estatística & dados numéricos , Carcinógenos/toxicidade , Exposição Ambiental/estatística & dados numéricos , Neoplasias/induzido quimicamente , Neoplasias/epidemiologia , Hidrocarbonetos Policíclicos Aromáticos/toxicidadeRESUMO
For genotoxic carcinogens, covalent binding to DNA is a critical initiating event in tumorigenesis. The present research investigated dose-effect relationships of three genotoxic carcinogens representing different structural classes, 2-acetylaminofluorene (2-AAF), benzo[a]pyrene (B[a]P) and quinoline (QUI), to assess the existence of no-observed-effect-levels (NOELs) for the formation of DNA adducts. Carcinogens were administered into the air sac of fertilized turkey eggs over wide dose ranges in three daily injections on days 22 to 24 of incubation. DNA adducts were measured in the fetal turkey livers by the 32P-nucleotide postlabeling (NPL) assay. B[a]P and QUI produced DNA adducts in a dosage-related manner and exhibited NOELs at 0.65 and 0.35 mg/kg bw/day, respectively. In contrast, 2-AAF formed DNA adducts at all tested dosages down to 0.005 mg/kg bw/day. Benchmark dose (BMD) analysis identified the potencies of 2-AAF and QUI to be similar, while B[a]P was the least potent compound. Overall, findings in fetal turkey livers demonstrated that exposure levels to genotoxic compounds that do not result in DNA adducts can exist but are not evident with all carcinogens of this type. The use of mechanistic dose-effect studies for genotoxic endpoints can provide critical information for prioritization of concerns for risk assessment.
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Carcinógenos , Adutos de DNA , Carcinógenos/metabolismo , Adutos de DNA/metabolismo , Fígado , Dano ao DNA , 2-Acetilaminofluoreno/farmacologia , 2-Acetilaminofluoreno/toxicidadeRESUMO
Pyrrolizidine alkaloids (PAs) and their N-oxides (PA-N-oxides) are phytotoxins found in food, feed and the environment. Yet, limited data exist from which the relative potency of a PA-N-oxide relative to its corresponding PA (REPPANO to PA) can be defined. This study aims to investigate the influence of dose, fraction bioactivated and endpoint on the REPPANO to PA of a series of pyrrolizidine N-oxides using in vitro-in silico data and physiologically based kinetic (PBK) modeling. The first endpoint used to calculate the REPPANO to PA was the ratio of the area under the concentration-time curve of PA resulting from an oral dose of PA-N-oxide divided by that from an equimolar dose of PA (Method 1). The second endpoint was the ratio of the amount of pyrrole-protein adducts formed under these conditions (Method 2). REPPANO to PA values appeared to decrease with increasing dose, with the decrease for Method 2 already starting at lower dose level than for Method 1. At dose levels as low as estimated daily human intakes, REPPANO to PA values amounted to 0.92, 0.81, 0.78, and 0.68 for retrorsine N-oxide, seneciphylline N-oxide, riddelliine N-oxide and senecivernine N-oxide, respectively, and became independent of the dose or fraction bioactivated, because no GSH depletion, saturation of PA clearance or PA-N-oxide reduction occurs. Overall, the results demonstrate the strength of using PBK modeling in defining REPPANO to PA values, thereby substantiating the use of the same approach for other PA-N-oxides for which in vivo data are lacking.
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In the context of the mysterious Balkan endemic nephropathy of the 1900s, and the discovery in the 1960s of the potent mycotoxin ochratoxin A, experimental research projects sought to explore any inter-relationship. Experimental lifetime administration of the toxin to male rats had revealed renal DNA adducts with the toxin, correlated with renal tumours, confirmation of which required molecular evidence. Consequently, production of 14C-ochratoxin A of a high specific radioactivity was required, practical biosynthetic detail of which had not previously been published. A fermentation study of Aspergillus ochraceous was carried out during 2002 for a European project, to select for the production of high-quality 14C-ochratoxin A, necessarily exploring for the maximum diversion of 14C-sodium acetate into the pentaketide portion of mycotoxin. Experimentation necessarily had to optimise the competitive context of fungal growth dynamics and addition of the biosynthetic precursor in the early days of shaken-flask fermentation before adding the radiolabelled precursor. From optimal fermentation, 50 mg of the 14C ochratoxin A was supplied within a European project for DNA adduct experimentation, but that proved negative as subsequently published. Experimental description of the radiolabelled ochratoxin A production was later made in a doctoral thesis, but is first publicised here. Further review of the literature reveals an explanation for the published failure to confirm rat DNA/ochratoxin A adduct formation, for which further experimentation is now recommended.
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Nefropatia dos Bálcãs , Micotoxinas , Ocratoxinas , Masculino , Animais , Ratos , Aspergillus ochraceus , FermentaçãoRESUMO
ABSTRACT Alcohol consumption has been linked to numerous pathologic conditions, including infectious diseases and several types of cancer. Alcohol exerts its modulatory effects on the immune system (IS) in a dose- and time-dependent manner. Numerous studies indicate that these alterations affect responses such as peripheral inflammation or decreased antibody production and promote chronic inflammation, leading to cell death. The molecular mechanisms underlying these effects involve generating an oxidative tissue environment, producing cell damage-associated molecular patterns (DAMPs), and activating pattern recognition receptors. In particular, toll-like receptors and their signaling system emerge as central elements whose activity is altered by alcohol intake. There is also some epidemiological evidence demonstrating the causal role of alcohol in the development of various types of cancer, such as head-and-neck cancer, esophageal cancer, colorectal cancer, liver cancer, and breast cancer. Most recent evidence suggests that factors related to alcohol consumption and cancer include increased levels of acetaldehyde, production of reactive oxygen species, alteration in DNA methylation, and modifications in retinoid metabolism. In addition, changes associated with alcohol use on the IS and intestinal microbiota may favor the growth of some types of tumors.
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Resumen A pesar de las grandes cantidades de formaldehído que se usa en la industria, la vigilancia de los efectos sobre la salud, derivados de la exposición ocupacional, sigue siendo un aspecto por mejorar en los programas de prevención de riesgos laborales. Objetivo: Identificar los diferentes biomarcadores de efecto que han sido propuestos para ser utilizados como indicadores indirectos de exposición a formaldehído y que se encuentran reportados en la literatura científica. Metodología: Se hizo una revisión bibliográfica desde el año 1990 hasta el primer trimestre del año 2018, usando los descriptores: "Formaldehído", "Biomarcador", "Exposición profesional". Se consultaron las bases de datos y se complementó la búsqueda inicial, mediante la consulta de la bibliografía citada por los autores de los artículos seleccionados. Se revisaron los resúmenes de los artículos y se seleccionaron aquellos que, luego de un análisis preliminar, se identificó contenían información relevante para la investigación. Resultados: Se encontraron 57 artículos relevantes para la revisión que contenían información sobre biomarcadores de efecto en poblaciones de trabajadores expuestos a formaldehído, algunos estudios en animales y células in vitro. Se resumieron los biomarcadores usados, listaron los estudios y describieron brevemente los hallazgos de cada uno. Conclusiones: La falta de conocimiento y entendimiento de los mecanismos toxicocinéticos y toxicodinámicos del formaldehído en los seres humanos limita la aplicación de un biomarcador de efecto que mida de manera práctica y sencilla la exposición al formaldehído exógeno.
Abstract Despite the large amounts of formaldehyde used in the industry, monitoring the health effects of occupational exposure remains an aspect of further improvement in occupational risk prevention programs. Objective: To identify the different biomarkers of effect that have been proposed to be used as indirect indicators of formaldehyde exposure and that are reported in the scientific literature. Methodology: A bibliographic review of literature from 1990 to the first quarter of 2018 was made using the descriptors: "Formaldehyde", "Biomarker", "Professional exposure". Databases were consulted and the initial search was supplemented by consulting the bibliography cited by the authors of the selected articles. Abstracts of articles were reviewed and those that, after preliminary analysis, were determined to contain information relevant to the research were selected. Results: The study found 57 articles relevant to the review which contained information on biomarkers of effect in populations of workers exposed to formaldehyde, some animal studies, and in vitro cells. Used biomarkers were summarized, studies were listed, and the findings of each of them were briefly described. Conclusions: The lack of knowledge and understanding of the toxicokinetic and toxicodynamic mechanisms of formaldehyde in humans limits the application of a biomarker of effect that measures in a practical and simple way exposure to exogenous formaldehyde.
Resumo Apesar da grande quantidade de formaldeído usado na indústria, o monitoramento dos efeitos na saúde, derivados da exposição ocupacional, continua sendo um aspecto a ser aprimorado nos programas de prevenção de riscos ocupacionais. Objetivo: Identificar os diferentes biomarcadores de efeito propostos para serem utilizados como indicadores indiretos da exposição ao formaldeído e relatados na bibliografia científica. Metodologia: Foi realizada uma revisão bibliográfica entre 1990 e o primeiro trimestre de 2018, utilizando os descritores: "Formaldeído", "Biomarcador", "Exposição Profissional". As bases de dados foram consultadas e a busca inicial foi complementada consultando a bibliografia citada pelos autores dos artigos selecionados. Os resumos dos artigos foram revisados e aqueles que, após análise preliminar, foram identificados, continham informações relevantes para a investigação. Resultados: Foram encontrados 57 artigos relevantes para a revisão que continham informações sobre biomarcadores de efeito em populações de trabalhadores expostos ao formaldeído, alguns estudos em animais e células in vitro. Os biomarcadores utilizados foram resumidos, os estudos listados e descritos brevemente os achados de cada um. Conclusões: A falta de conhecimento e entendimento dos mecanismos toxicocinéticos e toxicodinâmicos do formaldeído em humanos limita a aplicação de um biomarcador de efeito que mede de maneira prática e simples a exposição ao formaldeído exógeno.
RESUMO
The use of tobacco products is one of the established contributors toward the development and spread of oral cancer. Additionally, recent research has indicated oral microbiome, infections with Human papilloma virus (HPV), EpsteinBarr virus (EBV), Candida as significant contributing factors to this disease along with lifestyle habits. Deregulation of cellular pathways envisaging metabolism, transcription, translation, and epigenetics caused by these risk factors either individually or in unison is manifold, resulting in the increased risk of oral cancer. Globally, this cancer continues to exist as one of the major causes of cancer-related mortalities; the numbers in the developing South Asian countries clearly indicate yearly escalation. This review encompasses the variety of genetic modifications, including adduct formation, mutation (duplication, deletion, and translocation), and epigenetic changes evident in oral squamous cell carcinoma (OSCC). In addition, it highlights the interference caused by tobacco products in Wnt signaling, PI3K/Akt/mTOR, JAK-STAT, and other important pathways. The information provided also ensures a comprehensive and critical revisit to non-tobacco-induced OSCC. Extensive literature survey and analysis has been conducted to generate the chromosome maps specifically highlighting OSCC-related mutations with the potential to act as spectacles for the early diagnosis and targeted treatment of this disease cancer (AU)
Assuntos
Humanos , Infecções por Vírus Epstein-Barr/complicações , Neoplasias Bucais/genética , Neoplasias Bucais/virologia , Nicotiana/efeitos adversos , Mutação , Herpesvirus Humano 4/genética , Classe I de Fosfatidilinositol 3-Quinases/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço , Neoplasias Bucais/patologia , Fatores de RiscoRESUMO
Os aldeídos são espécies reativas que podem ser produzidos endogenamente por processos como a lipoperoxidação, podendo reagir com lipídios, proteínas e DNA. Diversas evidências apontam para o envolvimento de aldeídos reativos na progressão de patologias como doenças cardiovasculares, arteriosclerose e doenças neurodegenerativas. Uma meta central do CEPIDRedoxoma é estudar a reatividade química de intermediários redox em ambientes biológicos e consequentes mudanças na estrutura e função de biomoléculas, entender como cada intermediário redox reage com biomoléculas específicas e os efeitos resultantes, essenciais para a concepção de biomarcadores e antioxidantes. O nosso grupo estuda os mecanismos de formação, detoxificação e reação com biomoléculas de aldeídos reativos endógenos e exógenos e seu papel em patologias como a esclerose lateral amiotrófica (ALS). Um dos mecanismos de detoxificação desses aldeídos é através da conjugação com a carnosina. Recentemente, foi observado que a suplementação de animais transgênicos ALS SOD G93A com carnosina via oral resultou em retardo da perda de peso e tendência de aumento da sobrevida dos animais. O presente projeto buscou investigar o possível papel da carnosina em animais modelo para ALS. Para isso as modificações em DNA induzidas por aldeídos reativos e a formação de adutos de carnosina-aldeídos foram analisadas através de metodologia HPLC-MS/MS. Assim observamos que ratos suplementados com carnosina apresentaram níveis significativamente menores de proteína carbonilada em músculo e fígado. Em fígadoforam vistos níveis menores de dois adutos de DNA, 8-oxodGuo e1,N2-HO-propanodGuo, em animais suplementados. Em cérebro foram detectados níveis menores de 1, N2-εdGuo. Com relação aos adutos carnosina-aldeídos, foi observado níveis significativamente maiores do aduto CAR-HHE na medula. Com embasamento nos resultados aqui apresentados, sugere-se a utilização de sequestradores de aldeídos como uma estratégia terapêutica em condições fisiopatológicas nas quais ao acúmulo dessas espécies está comprovado
Aldehydes are reactive species that can be produced endogenously by processes such as lipid peroxidation, which can react with lipids, proteins and DNA. Several evidences point to the involvement of reactive aldehydes in the progression of pathologies such as cardiovascular diseases, atherosclerosis and neurodegenerative diseases. A central goal of CEPID-Redoxoma is to study the chemical reactivity of redox intermediates in biological environments and consequent changes in the structure and function of biomolecules, to understand how each redox intermediate reacts with specific biomolecules and the resulting effects, essential for the design of biomarkers and antioxidants. Our group studies the mechanisms of formation, detoxification and reaction with biomolecules of endogenous and exogenous reactive aldehydes and their role in pathologies such as amyotrophic lateral sclerosis (ALS). One of the detoxification mechanisms of these aldehydes is through carnosine conjugation. Recently, we observed that oral carnosine supplementation in transgenic ALS SODG93A animals resulted in delayed weight loss and a tendency to increase the survival of the animals. The present project investigated the potential role of carnosine in animal models for ALS. Thus, reactive aldehydes induced DNA modifications and carnosine aldehyde adducts were analyzed by HPLC-MS/MS. We observed that rats supplemented with carnosine presented significantly lower levels of protein carbonylation in muscle and liver. Lower levels of two DNA adducts, 8-oxodGuo and 1, N2-HO-propanodGuo, were observed in liver of the supplemented animals. Lower levels of 1, N2-εdGuo were detected in the brain. Regarding the carnosine-aldehydeadducts, significantly higher levels of the CAR-HHE adduct were observed in spinal cord. The results presented here suggest the use of aldehyde scavengers as a therapeutic strategy under pathological conditions in which is proven the accumulation of these species