| Full text | |
| Author(s): |
de Oliveira, Tiago Franco
;
Felcao de Oliveira, Antonio Anax
;
Lemos, Miriam
;
Veras, Mariana
;
Nascimento Saldiva, Paulo Hilario
;
Gennari de Medeiros, Marisa Helena
;
Di Mascio, Paolo
;
de Melo Loureiro, Ana Paula
Total Authors: 8
|
| Document type: | Journal article |
| Source: | JOVE-JOURNAL OF VISUALIZED EXPERIMENTS; v. N/A, n. 147, p. 19-pg., 2019-05-01. |
| Abstract | |
DNA adducts and oxidized DNA bases are examples of DNA lesions that are useful biomarkers for the toxicity assessment of substances that are electrophilic, generate reactive electrophiles upon biotransformation, or induce oxidative stress. Among the oxidized nucleobases, the most studied one is 8-oxo-7,8-dihydroguanine (8-oxoGua) or 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo), a biomarker of oxidatively induced base damage in DNA. Aldehydes and epoxyaldehydes resulting from the lipid peroxidation process are electrophilic molecules able to form mutagenic exocyclic DNA adducts, such as the etheno adducts 1,N-2-etheno-2'-deoxyguanosine (1,N-2-epsilon dGuo) and 1,N-6-etheno-2'deoxyadenosine (1,N-6-epsilon dAdo), which have been suggested as potential biomarkers in the pathophysiology of inflammation. Selective and sensitive methods for their quantification in DNA are necessary for the development of preventive strategies to slow down cell mutation rates and chronic disease development (e.g., cancer, neurodegenerative diseases). Among the sensitive methods available for their detection (high performance liquid chromatography coupled to electrochemical or tandem mass spectrometry detectors, comet assay, immunoassays, P-32-postlabeling), the most selective are those based on high performance liquid chromatography coupled to tandem mass spectrometry (HPLC-ESI-MS/MS). Selectivity is an essential advantage when analyzing complex biological samples and HPLC-ESI-MS/MS evolved as the gold standard for quantification of modified nucleosides in biological matrices, such as DNA, urine, plasma and saliva. The use of isotopically labeled internal standards adds the advantage of corrections for molecule losses during the DNA hydrolysis and analyte enrichment steps, as well as for differences of the analyte ionization between samples. It also aids in the identification of the correct chromatographic peak when more than one peak is present. We present here validated sensitive, accurate and precise HPLC-ESI-MS/MS methods that were successfully applied for the quantification of 8-oxodGuo, 1,N-6-dAdo and 1,N-2-dGuo in lung, liver and kidney DNA of A/J mice for the assessment of the effects of ambient PM2.5 exposure. (AU) | |
| FAPESP's process: | 13/07937-8 - Redoxome - Redox Processes in Biomedicine |
| Grantee: | Ohara Augusto |
| Support Opportunities: | Research Grants - Research, Innovation and Dissemination Centers - RIDC |
| FAPESP's process: | 12/22190-3 - The role of metabolic memory in the development of diabetic nephropathy |
| Grantee: | Ana Paula de Melo Loureiro |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 12/08616-8 - Characterization of a new biotransformation pathway of bisphenol A and quantitation of DNA lesions in HL-60 and MCF-7 cells |
| Grantee: | Ana Paula de Melo Loureiro |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 11/09891-0 - Genotoxic and epigenetic changes in DNA of mice exposed to particulate matter (PM 2,5) |
| Grantee: | Antonio Anáx Falcão de Oliveira |
| Support Opportunities: | Scholarships in Brazil - Scientific Initiation |
| FAPESP's process: | 12/08617-4 - Variables on glycemic control and pathogenesis of diabetic nephropathy: research on the phenomenon of metabolic memory |
| Grantee: | Antonio Anáx Falcão de Oliveira |
| Support Opportunities: | Scholarships in Brazil - Doctorate (Direct) |
| FAPESP's process: | 12/21636-8 - Transformation of human bronchial epithelial cells by benzo[a]pyrene: metabolic redox processes and DNA methylation / demethylation |
| Grantee: | Tiago Franco de Oliveira |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |