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1.
Aging (Albany NY) ; 15(14): 7258-7277, 2023 07 28.
Artículo en Inglés | MEDLINE | ID: mdl-37517089

RESUMEN

PURPOSE: Chronic inflammation and lipid peroxidation (LPO) are associated with the pathogenesis of hepatocellular carcinoma (HCC), and γ-hydroxy-1, N2-propanodeoxyguanosine (γ-OHPdG) is a promutagenic DNA adduct derived from LPO. This study aimed to examine the relationship between γ-OHPdG and the progression of liver carcinogenesis. METHODS: Primary HCC specimens were obtained from 228 patients and cirrhosis specimens from 46 patients. The patients were followed up with after surgery via outpatient visits and telephone calls. The levels of γ-OHPdG were determined by immunohistochemical analysis in the carcinomatous tissues together with adjacent and cirrhosis tissues. RESULTS: γ-OHPdG levels in the cancerous tissues were significantly higher compared to adjacent tissues (P < 0.001) and also higher than the ones from the tissues of cirrhosis patients. Along with tumor size, histological grade, MVI grade, T stage, the percentage of ki67-positive cells and HCC progression, γ-OHPdG levels in cancerous tissues showed a gradually increasing trend. Moreover, prognostic analysis showed that higher γ-OHPdG levels in cancerous tissues were strongly correlated with lower overall survival (P < 0.001), lower intrahepatic recurrence-free survival (P < 0.001) and lower distant metastasis-free survival (P < 0.05). There was a trend, although not statistically significant, of increased levels of γ-OHPdG in cirrhosis cases that advanced to HCC, whereas γ-OHPdG levels reversely correlated with the period of time observed for cirrhosis advanced to HCC. CONCLUSIONS: These results suggest that γ-OHPdG is a prognostic biomarker for predicting outcomes in HCC, and may serve as a prospective indicator for predicting HCC in cirrhosis patients.


Asunto(s)
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , Aductos de ADN , Pronóstico , Peroxidación de Lípido , Estudios Prospectivos , Cirrosis Hepática/diagnóstico , Cirrosis Hepática/complicaciones , Biomarcadores , Biomarcadores de Tumor/genética
2.
Int J Mol Sci ; 22(11)2021 Jun 02.
Artículo en Inglés | MEDLINE | ID: mdl-34199457

RESUMEN

Polycyclic aromatic hydrocarbons (PAHs) are chemical compounds comprised of carbon and hydrogen molecules in a cyclic arrangement. PAHs are associated with risks to human health, especially carcinogenesis. One form of exposure to these compounds is through ingestion of contaminated food, which can occur during preparation and processing involving high temperatures (e.g., grilling, smoking, toasting, roasting, and frying) as well as through PAHs present in the soil, air, and water (i.e., environmental pollution). Differently from changes caused by microbiological characteristics and lipid oxidation, consumers cannot sensorially perceive PAH contamination in food products, thereby hindering their ability to reject these foods. Herein, the occurrence and biological effects of PAHs were comprehensively explored, as well as analytical methods to monitor their levels, legislations, and strategies to reduce their generation in food products. This review updates the current knowledge and addresses recent regulation changes concerning the widespread PAHs contamination in several types of food, often surpassing the concentration limits deemed acceptable by current legislations. Therefore, effective measures involving different food processing strategies are needed to prevent and reduce PAHs contamination, thereby decreasing human exposure and detrimental health effects. Furthermore, gaps in literature have been addressed to provide a basis for future studies.


Asunto(s)
Carcinogénesis/efectos de los fármacos , Contaminación Ambiental/efectos adversos , Alimentos/efectos adversos , Hidrocarburos Policíclicos Aromáticos/efectos adversos , Benzopirenos/efectos adversos , Carcinogénesis/genética , Carbón Orgánico/efectos adversos , Culinaria , Aductos de ADN/efectos adversos , Análisis de los Alimentos , Manipulación de Alimentos , Humanos
3.
Methods Mol Biol ; 2279: 225-239, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33683698

RESUMEN

Aldehydes are abundantly present in tobacco smoke and in urban air pollution and are endogenously generated as products of the lipid peroxidation process. These molecules can react with DNA bases forming mutagenic exocyclic adducts, which have been used as biomarkers of aldehyde exposure and as potential tools for the study of inflammation, metal storage diseases, neurodegenerative disorders, and cancer. High-performance liquid chromatography-tandem mass spectrometry (HPLC/MS/MS) provides a highly precise, specific and ultrasensitive method for the detection of exocyclic DNA adducts. Here we present and describe a validated micro-HPLC-Electro Spray Ionization (ESI)-MS/MS method for the quantification of 1,N2-propanodGuo, an adduct produced following the reaction between 2'-deoxyguanosine and acetaldehyde or crotonaldehyde.


Asunto(s)
Aductos de ADN/metabolismo , Daño del ADN , Pulmón/metabolismo , Espectrometría de Masa por Ionización de Electrospray , Espectrometría de Masas en Tándem , Animales , Cromatografía Líquida de Alta Presión , Desoxiguanosina/metabolismo , Ratas
4.
Toxicol Sci ; 181(1): 115-124, 2021 04 27.
Artículo en Inglés | MEDLINE | ID: mdl-33566103

RESUMEN

The northern Gulf of Mexico has a long history of polycyclic aromatic hydrocarbon (PAH) contamination from anthropogenic activities, natural oil seepages, and the 2010 Deepwater Horizon explosion and oil spill. The continental shelf of the same area is a known breeding ground for sperm whales (Physeter macrocephalus). To evaluate PAH-DNA damage, a biomarker for potential cancer risk, we compared skin biopsies collected from Gulf of Mexico sperm whales in 2012 with skin biopsies collected from sperm whales in areas of the Pacific Ocean in 1999-2001. All samples were obtained by crossbow and comprised both epidermis and subcutaneous blubber. To evaluate exposure, 7 carcinogenic PAHs were analyzed in lipids extracted from Pacific Ocean sperm whale blubber, pooled by sex, and location. To evaluate PAH-DNA damage, portions of all tissue samples were formalin-fixed, paraffin-embedded, sectioned, and examined for PAH-DNA adducts by immunohistochemistry (IHC) using an antiserum elicited against benzo[a]pyrene-modified DNA, which crossreacts with several high molecular weight carcinogenic PAHs bound to DNA. The IHC showed widespread epidermal nuclear localization of PAH-DNA adducts in the Gulf of Mexico whales (n = 15) but not in the Pacific Ocean whales (n = 4). A standard semiquantitative scoring system revealed significantly higher PAH-DNA adducts in the Gulf of Mexico whales compared to the whales from the Pacific Ocean study (p = .0002).


Asunto(s)
Contaminación por Petróleo , Hidrocarburos Policíclicos Aromáticos , Contaminantes Químicos del Agua , Animales , Biopsia , Aductos de ADN , Monitoreo del Ambiente , Golfo de México , Humanos , Contaminación por Petróleo/análisis , Hidrocarburos Policíclicos Aromáticos/toxicidad , Cachalote , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/toxicidad
5.
J Toxicol Environ Health A ; 84(8): 331-344, 2021 04 18.
Artículo en Inglés | MEDLINE | ID: mdl-33372577

RESUMEN

This study aimed to investigate the relationship between endogenous antioxidant system, 8-hydroxydeoxyguanosine adduct (8-OHdG) repair, and apoptosis in mice treated with chromium(VI) alone and in the presence of the antigenotoxic compound (-)-epigallocatechin-3-gallate (EGCG). Groups of 5 Hsd:ICR male mice were divided and treated as follows: (1) control, vehicle only; (2) EGCG, 8.5 mg/kg by gavage alone; (3) CrO3, 20 mg/kg intraperitoneally alone; and (4) EGCG combined with CrO3, EGCG was administered 4 hr prior to CrO3. Peripheral blood parameters were analyzed before treatment administration (time 0), and 48 hr after exposure. The administration of EGCG increased 8-OHdG levels and superoxide dismutase (SOD) activity. Treatment with CrO3 increased number of micronucleus (MN) presence, elevated apoptotic/necrotic cells frequencies, decreased 8-OHdG levels, diminished total antioxidant capacity (TAC), increased glutathione (GSH) total levels, and lowered SOD activity. Administration of EGCG prior to treatment with CrO3 resulted in lower concentrations of MN, reduced apoptotic and necrotic cell number, and restored TAC and SOD activity to control levels. It is conceivable that the dose of EGCG plays an important role in the genotoxic damage protection pathways. Thus, this study confirms the action of EGCG as an antigenotoxic agent against chromium(VI)-induced oxidative insults and demonstrates potential protective pathways for EGCG actions to counteract genotoxic damage induced by this metal.


Asunto(s)
8-Hidroxi-2'-Desoxicoguanosina/metabolismo , Antimutagênicos/farmacología , Apoptosis , Catequina/análogos & derivados , Cromo/efectos adversos , Aductos de ADN/metabolismo , Contaminantes Ambientales/efectos adversos , Animales , Antioxidantes/metabolismo , Catequina/farmacología , Masculino , Ratones
6.
DNA Cell Biol ; 38(4): 307-313, 2019 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-30785779

RESUMEN

O6-Methylguanine-DNA methyltransferase (MGMT) is an enzyme that repairs the DNA damage caused by the tobacco habit, and low activity of this enzyme has been associated with a risk of lung cancer (LC). Our objective was to determine the association of the promoter methylation and the rs12917 polymorphism of MGMT with formation of DNA bulky adducts and the risk of LC in the Mexican Mestizo population. In this study are included 431 subjects. High-resolution melting analysis was used to determine the polymorphism MGMT rs12917 and methylation levels. DNA bulky adducts were determined by 32P-postlabeling. Our results showed that MGMT rs12917 and higher levels of methylation in the MGMT promoter are associated with the risk of LC. The levels of adducts are related with the phe/phe genotype and, only in the cases group, with the hypermethylation (>50%) of MGMT; however, this last association was not statistically significant.


Asunto(s)
Metilación de ADN , Metilasas de Modificación del ADN/genética , Enzimas Reparadoras del ADN/genética , Predisposición Genética a la Enfermedad/genética , Polimorfismo de Nucleótido Simple , Regiones Promotoras Genéticas/genética , Aspergilosis Pulmonar/genética , Proteínas Supresoras de Tumor/genética , Estudios de Casos y Controles , Aductos de ADN/metabolismo , Etnicidad/genética , Femenino , Humanos , Masculino , México/etnología , Persona de Mediana Edad , Aspergilosis Pulmonar/enzimología
7.
Environ Mol Mutagen ; 60(5): 428-442, 2019 06.
Artículo en Inglés | MEDLINE | ID: mdl-30706525

RESUMEN

Mexico City's Metropolitan Area (MCMA) includes Mexico City and 60 municipalities of the neighbor states. Inhabitants are exposed to emissions from over five million vehicles and stationary sources of air pollutants such as particulate matter (PM) and ozone. MCMA PM contains elemental carbon and organic carbon (OC). OCs include polycyclic aromatic hydrocarbons (PAHs), many of which induce mutagenic and carcinogenic DNA adducts. Gestational exposure to air pollution has been associated with increased risk of intrauterine growth restriction, preterm birth or low birth weight risk, and PAH-DNA adducts. These effects also depend on the presence of risk alleles. We investigated the presence of bulky PAH-DNA adducts, plasma 8-iso-PGF2α (8-iso-prostaglandin F2α ) and risk allele variants in neonates cord blood and their non-smoking mothers' leucocytes from families that were living in a highly polluted area during 2014-2015. The presence of adducts was significantly associated with both PM2.5 and PM10 levels, mainly during the last trimester of gestation in both neonates and mothers, while the last month of pregnancy was significant for the association between ozone levels and maternal plasma 8-iso-PGF2α . Fetal CYP1B1*3 risk allele was associated with increased adduct levels in neonates while the presence of the maternal allele significantly reduced the levels of fetal adducts. Maternal NQO1*2 was associated with lower maternal levels of adducts. Our findings suggest the need to reduce actual PM limits in MCMA. We did not observe a clear association between PM and/or adduct levels and neonate weight, length, body mass index, Apgar or Capurro score. Environ. Mol. Mutagen. 60:428-442, 2019. © 2019 Wiley Periodicals, Inc.


Asunto(s)
Aductos de ADN/análisis , Exposición Materna , Intercambio Materno-Fetal/fisiología , Ozono/toxicidad , Material Particulado/toxicidad , Hidrocarburos Policíclicos Aromáticos/análisis , Efectos Tardíos de la Exposición Prenatal/patología , Adulto , Contaminación del Aire/análisis , Citocromo P-450 CYP1B1/genética , Aductos de ADN/genética , Femenino , Sangre Fetal/química , Humanos , Recién Nacido , Isoprostanos/sangre , México , NAD(P)H Deshidrogenasa (Quinona)/genética , Embarazo , Emisiones de Vehículos/análisis , Adulto Joven
8.
DNA Repair (Amst) ; 73: 78-90, 2019 01.
Artículo en Inglés | MEDLINE | ID: mdl-30470509

RESUMEN

Genomes are affected by a wide range of damage, which has resulted in the evolution of a number of widely conserved DNA repair pathways. Most of these repair reactions have been described in the African trypanosome Trypanosoma brucei, which is a genetically tractable eukaryotic microbe and important human and animal parasite, but little work has considered how the DNA damage response operates throughout the T. brucei life cycle. Using quantitative PCR we have assessed damage induction and repair in both the nuclear and mitochondrial genomes of the parasite. We show differing kinetics of repair for three forms of DNA damage, and dramatic differences in repair between replicative life cycle forms found in the testse fly midgut and the mammal. We find that mammal-infective T. brucei cells repair oxidative and crosslink-induced DNA damage more efficiently than tsetse-infective cells and, moreover, very distinct patterns of induction and repair of DNA alkylating damage in the two life cycle forms. We also reveal robust repair of DNA lesions in the highly unusual T. brucei mitochondrial genome (the kinetoplast). By examining mutants we show that nuclear alkylation damage is repaired by the concerted action of two repair pathways, and that Rad51 acts in kinetoplast repair. Finally, we correlate repair with cell cycle arrest and cell growth, revealing that induced DNA damage has strikingly differing effects on the two life cycle stages, with distinct timing of alkylation-induced cell cycle arrest and higher levels of damage induced death in mammal-infective cells. Our data reveal that T. brucei regulates the DNA damage response during its life cycle, a capacity that may be shared by many microbial pathogens that exist in variant environments during growth and transmission.


Asunto(s)
Daño del ADN , Trypanosoma brucei brucei/crecimiento & desarrollo , Trypanosoma brucei brucei/genética , Alquilación , Puntos de Control del Ciclo Celular/genética , Aductos de ADN/metabolismo , Reparación del ADN , ADN Protozoario/genética , ADN Protozoario/metabolismo , Estrés Oxidativo/genética , Recombinasa Rad51/metabolismo , Trypanosoma brucei brucei/citología , Trypanosoma brucei brucei/metabolismo
9.
São Paulo; s.n; s.n; 2019. 93 p. tab, ilus, graf.
Tesis en Portugués, Francés | LILACS | ID: biblio-1025676

RESUMEN

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


Asunto(s)
Animales , Masculino , Femenino , Ratas , Fenómenos Biológicos , Carnosina/efectos adversos , Aldehídos/análisis , Esclerosis Amiotrófica Lateral/patología , Espectrometría de Masas/métodos , Cromatografía Liquida/métodos , Aductos de ADN
10.
Chem Res Toxicol ; 31(5): 332-339, 2018 05 21.
Artículo en Inglés | MEDLINE | ID: mdl-29707942

RESUMEN

Air pollution is a major environmental risk for human health. Acetaldehyde is present in tobacco smoke and vehicle exhaust. In this study, we show that [13C2]-acetaldehyde induces DNA modification with the formation of isotopically labeled 1, N2-propano-2'-deoxyguanosine adducts in the brain and lungs of rats exposed to concentrations of acetaldehyde found in the atmosphere of megacities. The adduct, with the addition of two molecules of isotopically labeled acetaldehyde [13C4]-1, N2-propano-dGuo, was detected in the lung and brain tissues of exposed rats by micro-HPLC/MS/MS. Structural confirmation of the products was unequivocally performed by nano-LC/ESI+-HRMS3 analyses. DNA modifications induced by acetaldehyde have been regarded as a key factor in the mechanism of mutagenesis and may be involved in the cancer risks associated with air pollution.


Asunto(s)
Acetaldehído/toxicidad , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Aductos de ADN/biosíntesis , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Acetaldehído/administración & dosificación , Acetaldehído/química , Animales , Isótopos de Carbono , Aductos de ADN/química , Aductos de ADN/aislamiento & purificación , Masculino , Estructura Molecular , Ratas , Ratas Wistar
11.
Biopolymers ; 107(12)2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-29058317

RESUMEN

In this work, we have studied the interaction between the anticancer drug doxorubicin (doxo) and condensed DNA, using optical tweezers. To perform this task, we use the protein bovine serum albumin (BSA) in the working buffer to mimic two key conditions present in the real intracellular environment: the condensed state of the DNA and the abundant presence of charged macromolecules in the surrounding medium. In particular, we have found that, when doxo is previously intercalated in disperse DNA, the drug hinders the DNA condensation process upon the addition of BSA in the buffer. On the other hand, when bare DNA is firstly condensed by BSA, doxo is capable to intercalate and to unfold the DNA condensates at relatively high concentrations. In addition, a specific interaction between BSA and doxo was verified, which significantly changes the chemical equilibrium of the DNA-doxo interaction. Finally, the presence of BSA in the buffer stabilizes the double-helix structure of the DNA-doxo complexes, preventing partial DNA denaturation induced by the stretching forces.


Asunto(s)
Antineoplásicos/química , Aductos de ADN/metabolismo , ADN/química , Doxorrubicina/química , Albúmina Sérica Bovina/química , Animales , Antineoplásicos/metabolismo , Bovinos , ADN/metabolismo , Doxorrubicina/metabolismo , Albúmina Sérica Bovina/metabolismo , Resonancia por Plasmón de Superficie , Temperatura
12.
Cell Stress Chaperones ; 22(6): 811-822, 2017 11.
Artículo en Inglés | MEDLINE | ID: mdl-28608263

RESUMEN

Neoadjuvant (or induction) chemotherapy can be used for cervical cancer patients with locally advanced disease; this treatment is followed by radical surgery and/or radiation therapy. Cisplatin is considered to be the most active platinum agent drug for this cancer, with a response rate of 20%. In order to understand how the cisplatin treatment affects the stress response, in this work, we performed an exploratory study to analyze a number of stress proteins before and after cisplatin neoadjuvant chemotherapy. The study involved 14 patients; the pre- and post-chemotherapy paired biopsies were examined by hematoxylin and eosin staining and by immunohistochemistry. The proteins evaluated were p53, P16/INK4A, MSH2, nuclear protein transcriptional regulator 1 (NUPR1), and HSPB1 (total: HSPB1/t and phosphorylated: HSPB1/p). These proteins were selected because there is previous evidence of their relationship with drug resistance. The formation of platinum-DNA adducts was also studied. There was a great variation in the expression levels of the mentioned proteins in the pre-chemotherapy biopsies. After chemotherapy, p53 was not significantly affected by cisplatin, as well as P16/INK4A and MSH2 while nuclear NUPR1 content tended to decrease (p = 0.056). Cytoplasmic HSPB1/t expression levels decreased significantly following cisplatin therapy while nuclear HSPB1/t and HSPB1/p tended to increase. Since the most significant changes following chemotherapy appeared in the HSPB1 expression levels, the changes were confirmed by Western blot. The platinum-DNA adducts were observed in HeLa cell in apoptosis; however, in the tumor samples, the platinum-DNA adducts were observed in morphologically healthy tumor cells; these cells displayed nuclear HSPB1/p. Further mechanistic studies should be performed to reveal how HSPB1/p is related with drug resistance. When the correlations of the markers with the response to neoadjuvant chemotherapy were examined, only high pre-chemotherapy levels of cytoplasmic HSPB1/p correlated with a poor clinical and pathological response to neoadjuvant cisplatin chemotherapy (p = 0.056) suggesting that this marker could be useful opening its study in a larger number of cases.


Asunto(s)
Biomarcadores de Tumor/genética , Cisplatino/efectos adversos , Proteínas de Choque Térmico HSP27/genética , Neoplasias del Cuello Uterino/tratamiento farmacológico , Adulto , Anciano , Cisplatino/administración & dosificación , Aductos de ADN/genética , Daño del ADN/efectos de los fármacos , Daño del ADN/genética , Reparación del ADN/efectos de los fármacos , Reparación del ADN/genética , Resistencia a Antineoplásicos/genética , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Células HeLa , Proteínas de Choque Térmico , Humanos , Persona de Mediana Edad , Chaperonas Moleculares , Terapia Neoadyuvante/efectos adversos , Proteína p53 Supresora de Tumor/genética , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/patología
13.
Biopolymers ; 107(3)2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-27718222

RESUMEN

We have investigated the interaction of the DNA molecule with the anticancer drug doxorubicin (doxo) by using three different experimental techniques: single molecule stretching, single molecule imaging, and dynamic light scattering. Such techniques allowed us to get new insights on the mechanical behavior of the DNA-doxo complexes as well as on the physical chemistry of the interaction. First, the contour length data obtained from single molecule stretching were used to extract the physicochemical parameters of the DNA-doxo interaction under different buffer conditions. This analysis has proven that the physical chemistry of such interaction can be modulated by changing the ionic strength of the surrounding buffer. In particular we have found that at low ionc strengths doxo interacts with DNA by simple intercalation (no aggregation) and/or by forming bound dimers. For high ionic strengths, otherwise, doxo-doxo self-association is enhanced, giving rise to the formation of bound doxo aggregates composed by 3 to 4 molecules along the double-helix. On the other hand, the results obtained for the persistence length of the DNA-doxo complexes is strongly force-dependent, presenting different behaviors when measured with stretching or non-stretching techniques.


Asunto(s)
Aductos de ADN/química , ADN/química , Doxorrubicina/química , ADN/metabolismo , Doxorrubicina/metabolismo , Dispersión Dinámica de Luz , Microscopía de Fuerza Atómica , Pinzas Ópticas
14.
Environ Res ; 152: 360-368, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27567517

RESUMEN

Environmental exposures along the US-Mexico border have the potential to adversely affect the maternal-fetal environment. The purpose of this study was to assess placental biomarkers of environmental exposures in an obstetric population at the California-Baja California border in relation to detoxifying enzymes in the placenta and nutritional status. This study was conducted on consenting, full-term, obstetric patients (n=54), delivering in a hospital in Tijuana, Baja California (BC), Mexico. Placental polyaromatic hydrocarbon (PAH)-DNA adducts were measured in addition to placental glutathione-S-transferase (GST) activity and genotype, maternal serum folate, and maternal and umbilical cord blood lead and cadmium levels. A questionnaire was administered to the mothers to determine maternal occupation in a maquiladora, other exposures, and obstetric indicators. In univariate analysis, maternal serum folate levels were inversely correlated with total PAH-DNA adducts (rho=-0.375, p=0.007); adduct #1 (rho=-0.388, p=0.005); and adduct #3 (rho =-0.430, p=0.002). Maternal lead levels were significantly positively correlated with cord blood lead levels (rho=0.512, p<0.001). Cadmium levels were generally very low but significantly higher in mothers exposed to environmental tobacco smoke (ETS) (either at work or at home, n=10). In multivariate analysis, only maternal serum folate levels remained as a significant negative predictor of total DNA-PAH adducts levels in placenta. These findings affirm that placental tissue is a valuable and readily available source of human tissue for biomonitoring; and indicate that further study of the role of nutrition in detoxification and mitigation of environmental exposures in pregnant women is warranted.


Asunto(s)
Contaminantes Ambientales/metabolismo , Glutatión Transferasa/genética , Placenta/química , Adolescente , Adulto , Biomarcadores/metabolismo , Cadmio/metabolismo , Aductos de ADN/genética , Aductos de ADN/metabolismo , Femenino , Glutatión Transferasa/metabolismo , Humanos , Plomo/metabolismo , México , Estado Nutricional , Placenta/enzimología , Hidrocarburos Policíclicos Aromáticos/metabolismo , Embarazo , Adulto Joven
15.
Mutat Res Rev Mutat Res ; 768: 68-77, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27234564

RESUMEN

In cancer treatment, chemotherapy is one of the main strategies used. The knowledge of the cellular and molecular characteristics of tumors allows the use of more specific drugs, making the removal of tumors more efficient. Among the drugs of choice in these treatments, topoisomerase inhibitors are widely used against different types of tumors. Topoisomerases are enzymes responsible for maintaining the structure of DNA, altering its topological state temporarily during the processes of replication and transcription, in order to avoid supercoiling and entanglements at the double helix. The DNA damage formed as a result of topoisomerase inhibition can be repaired by DNA repair mechanisms. Thus, DNA repair pathways can modulate the effectiveness of chemotherapy. Homologous recombination (HR) and non-homologous end joining (NHEJ) are the main pathways involved in the removal of double strand breaks (DSBs); while nucleotide excision repair (NER) is mainly characterized by the removal of lesions that lead to significant structural distortions in the DNA double helix. Evidence has shown that DSBs are the main type of damage resulting from the inhibition of the DNA topoisomerase II enzyme, and therefore the involvement of HR and NHEJ pathways in the repair process is well established. However, some topoisomerase II inhibitors induce other types of lesions, like DNA adducts, interstrand crosslinks and reactive oxygen species, and studies have shown that other DNA repair pathways might be participating in removing injury induced by these drugs. This review aims to correlate the involvement of proteins from different DNA repair pathways in response to these drugs, with an emphasis on NER.


Asunto(s)
Daño del ADN/efectos de los fármacos , Reparación del ADN , ADN-Topoisomerasas/metabolismo , Inhibidores de Topoisomerasa II/farmacología , Animales , Aductos de ADN , Humanos , Oxidación-Reducción , Especies Reactivas de Oxígeno , Inhibidores de Topoisomerasa I/química , Inhibidores de Topoisomerasa I/farmacología , Inhibidores de Topoisomerasa II/química
16.
São Paulo; s.n; 2016. 115 p.
Tesis en Portugués | LILACS | ID: biblio-868246

RESUMEN

Introdução. O alto consumo de carne, principalmente vermelha e processada, tem sido relacionado com aumento de risco de doenças crônicas, especialmente o câncer. Uma das explicações possíveis são os métodos de preparo culinário a altas temperaturas, que acarretam na formação aminas heterocíclicas. Estes compostos são detoxificados no nosso organismo, passando por um processo, no qual podem ser geradas espécies reativas, relacionadas ao estresse oxidativo e ao dano ao DNA. Entretanto, os indivíduos apresentam respostas diferentes à mesma exposição dietética, podendo ter diferentes níveis de risco ou benefício com a mesma ingestão de alimentos. O código genético individual pode ser uma das causas dessa variação interpessoal. Objetivo. Investigar a relação entre o consumo de carnes e aminas heterocíclicas com estresse oxidativo e dano no DNA, considerando polimorfismos genéticos, fatores demográficos e de estilo de vida em residentes do Município de São Paulo. Métodos. Foram utilizados dados dietéticos, genéticos, bioquímicos e estilo de vida de um estudo transversal com amostra probabilística de múltiplo estágio chamado Inquérito de Saúde de São Paulo (ISACapital). Os dados de carne e aminas heterocíclicas foram obtidos a partir de um recordatório alimentar de 24 horas e questionário sobre métodos de cocção e graus de cozimento das carnes. A extração do DNA ocorreu pelo método por sal e utilizou-se a técnica PCR em tempo real para determinação dos seguintes polimorfismos de nucleotídeo único: CYP1A1 (rs1048943), CYP1A2 (rs762551, rs35694136), CYP1B1 (rs1056836, rs10012), NAT2 (rs1208, rs1041983, rs1799929, rs1801280, rs1799931, rs1799930, rs1801279), NAT1 (rs4986782, rs5030839, rs56379106, rs56318881, rs6586714), SULT1A1 (rs928286), UGT1A9 (rs3832043), SOD2 (rs4880), CAT (rs7943316), GSTA1 (rs3957357), GSTP1 (rs1695), e deleção dos genes GSTM1 e GSTT1. Foram utilizados os biomarcadores malonaldeído (MDA) no plasma para estimar o estresse oxidativo e o 8-OHdG no plasma para estimar dano ao DNA. As associações foram examinadas por meio de modelos de regressão múltipla linear e logística ajustadas por sexo, idade, IMC, consumo de frutas e calorias, atividade física e fumo. Resultados. O consumo médio de aminas heterocíclicas foi de 437ng/dia e a carne de boi foi a que mais contribuiu para o consumo de aminas. Participantes que consumiram carne de boi grelhada muito bem passada apresentaram maiores concentrações de MDA do que os demais. Encontrou-se associação positiva entre consumo de aminas heterocíclicas com estresse oxidativo e dano ao DNA, isto é, indivíduos que consumiram maiores teores de aminas heterocíclicas apresentaram maiores chances de ter elevados concentrações de MDA (OR=1,17; P=0,04) e maiores concentrações de 8-OHdG (=1,62; P=0,04). Observou-se também que esta associação pode ser modificada pelas características genéticas individuais, sendo que polimorfismos nos genes das enzimas de detoxificação NAT2 e CYP1B1 interagiram com o consumo de aminas, diminuindo o estresse oxidativo. Conclusão. Verificou-se que o alto consumo de aminas heterocíclicas contribuiu para maiores níveis de estresse oxidativo e dano ao DNA independente de fatores demográficos e de estilo de vida, aumentando o risco de doenças crônicas. Observou-se também que esta relação pode ser alterada na presença de polimorfismos genéticos individuais


Introduction. The excessive meat intake, especially red and processed meat, has been linked to chronic diseases, especially cancer. One of the reasons for that is the cooking process at high temperatures that can form heterocyclic amines (HCA). During HCA metabolism, reactive species can be formed, which can cause oxidative stress and DNA damage. However, people can show different answers to the same food intake, increasing or decreasing the risk of diseases. The DNA code can be one of the causes of this between-person variations. Objective. To investigate the association between meat/heterocyclic amine intake with oxidative stress and DNA damage, considering polymorphism, demographic and life style factors among population of São Paulo city. Methods. Information on food intake, genetics, biochemical, and lifestyle was obtained from a representative, multistage probability-based cross-sectional study titled Health Survey for Sao Paulo (ISA-Capital). Meat and heterocyclic amine intake was estimated by a 24-hour dietary recall complemented by a detailed questionnaire with preferences of cooking methods and level of doneness for meats. The salt method was used for DNA extraction and real time PCR to identify the following single nucleotide polymorphisms: CYP1A1 (rs1048943), CYP1A2 (rs762551, rs35694136), CYP1B1 (rs1056836, rs10012), NAT2 (rs1208, rs1041983, rs1799929, rs1801280, rs1799931, rs1799930, rs1801279), NAT1 (rs4986782, rs5030839, rs56379106, rs56318881, rs6586714), SULT1A1 (rs928286), UGT1A9 (rs3832043), SOD2 (rs4880), CAT (rs7943316), GSTA1 (rs3957357), GSTP1 (rs1695), GSTM1 and GSTT1 (null or not). We used malondialdehyde (MDA) concentration in plasma to estimated oxidative stress, and 8-OHdG concentration in plasma to estimate DNA damage. Analyses were performed using multivariate logistic and linear regressions adjusted for smoking, sex, age, body mass index, energy intake, fruit intake, smoking and physical activity. Results. Mean HCA intake was 437ng/day and beef was the meat that contributed more to HCA. Participants who consumed grilled beef very well-done presented more MDA concentration than other participants. We found significant association between heterocyclic amine intake with oxidative stress and DNA damage. Participants who consumed high levels of heterocyclic amines showed higher odds to show high MDA concentration (OR=1.17; P=0.04) and high 8-OHdG concentration (=1.62; P=0.04). These associations could be modified by individual genetic characteristics. Polymorphisms in genes that codify NAT2 and CYP1B1 detoxification enzymes interacted with HCA intake, decreasing oxidative stress. Conclusions. The high heterocyclic amine intake contributed to increase oxidative stress independently of lifestyle and demographic factors, increasing risk of chronic diseases. These relationships can be modified by genetic polymorphisms


Asunto(s)
Aminas/química , Aductos de ADN , Ingestión de Alimentos , Carne/estadística & datos numéricos , Estrés Oxidativo/fisiología , Estudios Transversales , Dados Estadísticos , Compuestos Heterocíclicos/química , Estilo de Vida , Polimorfismo Genético , Factores de Riesgo
17.
Environ Pollut ; 203: 175-182, 2015 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-25909326

RESUMEN

The carcinogenic potential of urban particulate matter (PM) has been partly attributed to polycyclic aromatic hydrocarbons (PAHs) content, which activates the aryl hydrocarbon receptor (AhR). Here we report the effect of PM with an aerodynamic size of 10 µm (PM10) on the induction of AhR pathway in A549 cells, evaluating its downstream targets CYP1B1, IL-6, IL-8 and c-Jun. Significant increases in CYP1B1 protein and enzyme activity; IL-6 and IL-8 secretion and c-Jun protein were found in response to PM10. The formation of PAH-DNA adducts was also detected. The involvement of AhR pathway was confirmed with Resveratrol as AhR antagonist, which reversed CYP1B1 and c-Jun induction. Nevertheless, in IL-6 and IL-8 secretion, the Resveratrol was ineffective, suggesting an effect independent of this pathway. Considering the role of c-Jun in oncogenesis, its induction by PM may be contributing to its carcinogenic potential through induction of AhR pathway by PAHs present in PM10.


Asunto(s)
Contaminantes Atmosféricos/toxicidad , Material Particulado/toxicidad , Hidrocarburos Policíclicos Aromáticos/toxicidad , Proteínas Proto-Oncogénicas c-jun/metabolismo , Línea Celular Tumoral , Ciudades , Citocromo P-450 CYP1B1/metabolismo , Aductos de ADN/metabolismo , Humanos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , México , Hidrocarburos Policíclicos Aromáticos/metabolismo , Receptores de Hidrocarburo de Aril/antagonistas & inhibidores , Receptores de Hidrocarburo de Aril/metabolismo , Resveratrol , Estilbenos/farmacología
18.
Hist. ciênc. saúde-Manguinhos ; Hist. ciênc. saúde-Manguinhos;21(4): 1093-1112, Oct-Dec/2014.
Artículo en Portugués | LILACS | ID: lil-732505

RESUMEN

O objetivo deste trabalho é analisar o discurso do excesso sexual produzido pelo pensamento social brasileiro das décadas de 1920 e 1930 na sua interlocução com o discurso médico da época. De inspiração foucaultiana, o texto inscreve-se no campo da história dos saberes e está subsidiado por documentos sociológicos e médicos do período de referência. No quadro da recodificação vintecentista sobre o imaginário da brasilidade, o tema do excesso sexual foi revisitado pelo pensamento sociológico local, forjando-o ora como um perturbador do projeto civilizatório nacional, ora como um traço que deveria ser positivado por ter sido a condição de possibilidade da hibridização cultural de suas matrizes identitárias.


The objective of this article is to analyze the discourse of sexual excess produced by Brazilian social thinking in the 1920s and 1930s and its dialog with the medical discourse at the time. Inspired by Foucault, it is within the field of the history of knowledge and is supported by sociology and medical documents from the period in question.Within the framework of the twentieth century re-codification of the imagery of Brazilianness, the topic of sexual excess was revisited by local thinkers in the field of sociology and seen either as disturbing the national civilizing project, or as a trait that should be seen in a positive light because it permitted the cultural hybridization of its sources of identity.


Asunto(s)
Animales , Bovinos , Adenina/metabolismo , Aductos de ADN/metabolismo , Compuestos Epoxi/metabolismo , Guanina/metabolismo , Mutágenos/metabolismo , Aductos de ADN/química , Aductos de ADN/aislamiento & purificación , Compuestos Epoxi/química , Compuestos Epoxi/aislamiento & purificación , Guanosina/metabolismo , Mutágenos/química , Mutágenos/aislamiento & purificación
19.
Rev. panam. salud pública ; 36(5): 290-299, nov. 2014. tab
Artículo en Inglés | LILACS | ID: lil-733231

RESUMEN

OBJECTIVE: To examine the prevalence of blindness, visual impairment, and related eye diseases and conditions among adults in El Salvador, and to explore socioeconomic inequalities in their prevalence by education level and occupational status, stratified by sex. METHODS: Based upon the Rapid Assessment of Avoidable Blindness (RAAB) methodology, this nationwide sample comprised 3 800 participants (3 399 examined) ≥ 50 years old from 76 randomly selected clusters of 50 persons each. The prevalence of blindness, visual impairment and related eye diseases and conditions, including uncorrected refractive error (URE), was calculated for categories of education level and occupational status. Multiple logistic regression models were fitted to calculate odds ratios (ORs) and 95% confidence intervals (CIs) and stratified by sex. RESULTS: Age-adjusted prevalence was 2.4% (95% CI: 2.2-2.6) for blindness (men: 2.8% (95% CI: 2.5-3.1); women: 2.2% (95% CI: 1.9-2.5)) and 11.8% (95% CI: 11.6-12.0) for moderate visual impairment (men: 10.8% (95% CI: 10.5-11.1); women: 12.6% (95% CI: 12.4-12.8)). The proportion of visual impairment due to cataract was 43.8% in men and 33.5% in women. Inverse gradients of socioeconomic inequalities were observed in the prevalence of visual impairment. For example, the age-adjusted OR (AOR) was 3.4 (95% CI: 2.0-6.4) for visual impairment and 4.3 (95% CI: 2.1-10.4) for related URE in illiterate women compared to those with secondary education, and 1.9 (95% CI: 1.1-3.1) in cataract in unemployed men. CONCLUSIONS: Blindness and visual impairment prevalence is high in the El Salvador adult population. The main associated conditions are cataract and URE, two treatable conditions. As socioeconomic and gender inequalities in ocular health may herald discrimination and important barriers to accessing affordable, good-quality, and timely health care services, prioritization of public eye health care and disability policies should be put in place, particularly among women, the unemployed, and uneducated people.


OBJETIVO: Analizar la prevalencia de la ceguera, la deficiencia visual, y las enfermedades y afecciones oculares relacionadas en adultos de El Salvador, y explorar las desigualdades socioeconómicas en cuanto a su prevalencia según el nivel educativo y la situación laboral, estratificados por sexos. MÉTODOS: Se adoptó el método de Evaluación Rápida de la Ceguera Evitable, y se escogió una muestra a escala nacional de 3 800 participantes (de ellos se examinaron 3 399) de 50 años de edad o mayores, pertenecientes a 76 agrupamientos seleccionados aleatoriamente y constituidos por 50 personas cada uno. Se calculó la prevalencia de la ceguera, la deficiencia visual y las enfermedades y afecciones oculares relacionadas, incluido el error de refracción no corregido, según las diferentes categorías de nivel educativo y situación laboral. Se emplearon modelos de regresión logística múltiple para calcular las razones de posibilidades (OR) y los intervalos de confianza (IC) de 95%, y se estratificaron por sexos. RESULTADOS: La prevalencia ajustada por edad fue de 2,4% (IC de 95%: 2,2-2,6) para la ceguera (hombres: 2,8% [IC de 95%: 2,5-3,1]; mujeres: 2,2% [IC de 95%: 1,9-2,5]) y de 11,8% (IC de 95%: 11,6-12,0) para la deficiencia visual moderada (hombres: 10,8% [IC de 95%: 10,5-11,1]; mujeres: 12,6% [IC de 95%: 12,4-12,8]). La proporción de deficiencias visuales debidas a catarata fue de 43,8% en los hombres y de 33,5% en las mujeres. En la prevalencia de la deficiencia visual se observaron gradientes inversos de desigualdades socioeconómicas. Por ejemplo, la OR ajustada por edad fue de 3,4 (IC de 95%: 2,0-6,4) para la deficiencia visual y de 4,3 (IC de 95%: 2,1-10,4) para el error de refracción no corregido relacionado en las mujeres analfabetas, en comparación con las que tenían un nivel de educación secundaria, y fue de 1,9 (IC de 95%: 1,1-3,1) para la catarata en los hombres desempleados. CONCLUSIONES: La prevalencia de ceguera y deficiencia visual es alta en la población adulta de El Salvador. Las principales afecciones asociadas son la catarata y el error de refracción no corregido, ambas tratables. Puesto que las desigualdades socioeconómicas y de género en materia de salud ocular pueden ser indicativas de discriminación y de la existencia de barreras importantes para obtener acceso a servicios de atención de salud asequibles, de buena calidad y oportunos, es preciso dar prioridad a la atención oftalmológica pública y a las políticas dirigidas a corregir la discapacidad, en particular en las mujeres y en las personas desempleadas y sin formación.


Asunto(s)
Carcinógenos/química , Carcinógenos/síntesis química , Aductos de ADN/biosíntesis , Aductos de ADN/química , Compuestos Epoxi/química , Compuestos Epoxi/síntesis química , Guanosina/química , Aductos de ADN/efectos de los fármacos , Estabilidad de Medicamentos , Compuestos Epoxi/toxicidad , Cinética , Espectrometría de Masas , Estereoisomerismo
20.
Rev. panam. salud pública ; 36(5): 324-330, nov. 2014. ilus, tab
Artículo en Portugués | LILACS | ID: lil-733235

RESUMEN

OBJETIVO: Revisar as experiências de atenção fisioterapêutica dirigidas à população pediátrica descritas na literatura e analisar a produção de conhecimento sobre fisioterapia no contexto da atenção primária à saúde infantil (APSI). MÉTODOS: Foi realizada uma revisão sistemática conforme PRISMA, com busca nas seguintes bases de dados: MEDLINE, LILACS, SciELO, PubMed, Scopus, Cochrane; banco de teses da CAPES; e System for Information on Grey Literature in Europe (SIGLE). Foram utilizados os descritores "atenção primária à saúde", "fisioterapia", "lactente ou criança" e seus correspondentes na língua inglesa e espanhola, sem restrição de ano de publicação. RESULTADOS: Analisamos 13 artigos de seis países, reunidos em três eixos temáticos: dilemas profissionais (três artigos), competências e habilidades específicas para a APSI (sete artigos) e relatos de prática (quatro artigos). Os dilemas profissionais mencionados foram a ampliação do papel do fisioterapeuta para incluir ambientes comunitários, compartilhando a tomada de decisão com as famílias, e o trabalho em colaboração com outros serviços de saúde para identificar as necessidades da criança. As competências e habilidades citadas foram a identificação de sintomas clínicos e socioculturais para além das condições musculoesqueléticas, o diagnóstico fisioterapêutico precoce, a prevenção contra o uso excessivo de medicamentos e a capacidade de trabalhar em equipe. Os relatos de prática discorreram sobre estimulação em crianças com quadros neurológicos, tratamento respiratório e grupos com mães de crianças com esses acometimentos. CONCLUSÕES: O baixo número de estudos sugere desconhecimento quanto ao modo como a fisioterapia se insere na APSI e, provavelmente, quanto às habilidades profissionais necessárias nesse ambiente. Assim, são necessários mais estudos para fornecer dados sobre a área e um esforço de qualificação continuada por parte dos fisioterapeutas.


OBJECTIVE: To review pediatric physical therapy experiences described in the literature and to analyze the production of knowledge on physical therapy in the context of pediatric primary health care (PPHC). METHODS: A systematic review was conducted according to the PRISMA criteria. The following databases were searched: MEDLINE, LILACS, SciELO, PubMed, Scopus and Cochrane; Brazilian Ministry of Health's CAPES doctoral dissertations database; and System for Information on Grey Literature in Europe (SIGLE). The following search terms were used: ["primary health care" and ("physical therapy" or "physiotherapy") and ("child" or "infant")] and equivalent terms in Portuguese and Spanish, with no restriction on publication year. RESULTS: Thirteen articles from six countries were analyzed and grouped into three main themes: professional dilemmas (three articles), specific competencies and skills required in a PPHC setting (seven articles), and practice reports (four articles). Professional dilemmas involved expanding the role of physical therapists to encompass community environments and sharing the decision-making process with the family, as well as collaborative work with other health services to identify the needs of children. The competencies and skills mentioned in the literature related to the identification of clinical and sociocultural symptoms that go beyond musculoskeletal conditions, the establishment of early physical therapy diagnoses, prevention of overmedication, and the ability to work as team players. Practice reports addressed stimulation in children with neurological diseases, respiratory treatment, and establishing groups with mothers of children with these conditions. CONCLUSIONS: The small number of studies identified in this review suggests that there is little knowledge regarding the roles of physical therapists in PPHC and possibly regarding the professional abilities required in this setting. Therefore, further studies are required to provide data on the field, along with a continuing education effort on the part of physical therapists.


Asunto(s)
Adenosina/análogos & derivados , Compuestos Epoxi/química , Inosina/química , Mutágenos/química , Adenosina/química , Cromatografía Líquida de Alta Presión , Aductos de ADN/química , Inosina/análogos & derivados , Cinética , Espectroscopía de Resonancia Magnética , Espectrometría de Masa Bombardeada por Átomos Veloces , Espectrofotometría Ultravioleta
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