RESUMEN
BACKGROUND: Fibrotic Interstitial lung diseases (ILD) are a heterogeneous group of chronic lung diseases characterized by diverse degrees of lung inflammation and remodeling. They include idiopathic ILD such as idiopathic pulmonary fibrosis (IPF), and ILD secondary to chronic inflammatory diseases such as connective tissue disease (CTD). Precise differential diagnosis of ILD is critical since anti-inflammatory and immunosuppressive drugs, which are beneficial in inflammatory ILD, are detrimental in IPF. However, differential diagnosis of ILD is still difficult and often requires an invasive lung biopsy. The primary aim of this study is to identify volatile organic compounds (VOCs) patterns in exhaled air to non-invasively discriminate IPF and CTD-ILD. As secondary aim, the association between the IPF and CTD-ILD discriminating VOC patterns and functional impairment is investigated. METHODS: Fifty-three IPF patients, 53 CTD-ILD patients and 51 controls donated exhaled air, which was analyzed for its VOC content using gas chromatograph- time of flight- mass spectrometry. RESULTS: By applying multivariate analysis, a discriminative profile of 34 VOCs was observed to discriminate between IPF patients and healthy controls whereas 11 VOCs were able to distinguish between CTD-ILD patients and healthy controls. The separation between IPF and CTD-ILD could be made using 16 discriminating VOCs, that also displayed a significant correlation with total lung capacity and the 6 min' walk distance. CONCLUSIONS: This study reports for the first time that specific VOC profiles can be found to differentiate IPF and CTD-ILD from both healthy controls and each other. Moreover, an ILD-specific VOC profile was strongly correlated with functional parameters. Future research applying larger cohorts of patients suffering from a larger variety of ILDs should confirm the potential use of breathomics to facilitate fast, non-invasive and proper differential diagnosis of specific ILDs in the future as first step towards personalized medicine for these complex diseases.
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Aire/análisis , Pruebas Respiratorias/métodos , Espiración , Enfermedades Pulmonares Intersticiales/metabolismo , Capacidad Vital/fisiología , Adulto , Anciano , Anciano de 80 o más Años , Diagnóstico Diferencial , Femenino , Humanos , Enfermedades Pulmonares Intersticiales/diagnóstico , Masculino , Persona de Mediana Edad , Proyectos Piloto , Estudios Prospectivos , Tomografía Computarizada por Rayos XRESUMEN
Genotoxic carcinogens significantly damage cells and tissues by targeting macromolecules such as proteins and DNA, but their mechanisms of action and effects on human health are diverse. Consequently, determining the amount of exposure to a carcinogen and its cellular effects is essential, yet difficult. The aim of this manuscript was to investigate the potential of detecting alterations in volatile organic compounds (VOCs) profiles in the in vitro headspace of pulmonary cells after exposure to the genotoxic carcinogens cisplatin and benzo[a]pyrene using two different sampling set-ups. A prototype set-up was used for the cisplatin exposure, whereas a modified set-up was utilized for the benzo[a]pyrene exposure. Both carcinogens were added to the cell medium for 24 h. The headspace in the culture flask was sampled to measure the VOC content using gas chromatography-time-of-flight-mass spectrometry. Eight cisplatin-specific VOCs and six benzo[a]pyrene-specific VOCs were discriminatory between treated and non-treated cells. Since the in vivo biological effects of both genotoxic compounds are well-defined, the origin of the identified VOCs could potentially be traced back to common cellular processes including cell cycle pathways, DNA damage and repair. These results indicate that exposing lung cells to genotoxins alters headspace VOC profiles, suggesting that it might be possible to monitor VOC changes in vivo to study drug efficacy or exposure to different pollutants. In conclusion, this study emphasizes the innovative potential of in vitro VOCs experiments to determine their in vivo applicability and discover their endogenous origin.
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Mutágenos/toxicidad , Compuestos Orgánicos Volátiles/análisis , Células A549 , Benzo(a)pireno/toxicidad , Cisplatino/toxicidad , Daño del ADN , Cromatografía de Gases y Espectrometría de Masas , Humanos , Análisis de Componente PrincipalRESUMEN
Two biomarkers of exposure to cigarette smoke, 4-aminobiphenyl-hemoglobin (Hb) adducts and aromatic DNA adducts in lymphocytes, were determined from a population of 55 smokers and 4 nonsmokers. The levels of these adducts were related to daily cigarette consumption and also to (calculated) tar and nicotine intake. The Hb adduct levels seemed to correspond best to the number of cigarettes (cig) smoked, but at a cigarette consumption of >30 cig/day, a saturation effect was observed. Lymphocytic DNA adducts also correlated well with cigarette and tar consumption; for this type of adduct, a saturation level was reached at a dose of approximately 15-20 cig/day. From a subpopulation, a second sample was obtained after 2 months, and the adduct levels were compared with their initial adduct levels. Strong correlations were found between the first and second DNA adduct measurements (r = 0.84). In another subpopulation, resampling was performed after 6 months. No correlation between DNA adduct levels in the first and last samples was found, but 4-aminobiphenyl Hb adduct levels were strongly correlated (r = 0.78), the absolute quantities measured being comparable (paired t test: t = -1.27, P = 0.22, n = 15). We found no influence of GSTM1 and NAT2 polymorphisms on Hb adduct formation and of GSTM1 polymorphism on aromatic DNA adduct formation. A significantly lower aromatic DNA adduct level was observed for intermediate acetylators when compared to slow acetylators.
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Compuestos de Aminobifenilo/análisis , Aductos de ADN/análisis , Hemoglobina A/análisis , Linfocitos/química , Fumar/sangre , Adulto , Compuestos de Aminobifenilo/sangre , Arilamina N-Acetiltransferasa/genética , Biomarcadores/análisis , Biomarcadores/sangre , Aductos de ADN/sangre , Femenino , Glutatión Transferasa/genética , Humanos , Linfocitos/metabolismo , Masculino , Fenotipo , Polimorfismo Genético , Análisis de RegresiónRESUMEN
Gastric juice samples of 71 patients undergoing upper gastrointestinal endoscopy were collected as well as saliva samples from 40 of these patients. Age, sex, endoscopic diagnosis and medication were recorded. The gastric juice samples were analyzed for the presence and quantity of individual volatile N-nitrosamines, which were detected by gas chromatography-mass spectrometry, without prior derivatization. The samples were screened for eight nitrosamines, i.e. N-nitrosodimethylamine, N-nitrosoethylmethylamine, N-nitrosodiethylamine, N-nitrosodi-n-propylamine, N-nitrosodi-n-butylamine, N-nitrosopyrrolidine, N-nitrosopiperidine and N-nitrosomorpholine. The pH of the fresh gastric juice as well as nitrate and nitrite levels of gastric juice and saliva were determined. The mean total level of volatile N-nitrosamines in gastric juice was found to be 4.84 nmol/l (range 0-17.7 nmol/l). The main N-nitrosamines found were N-nitrosodiethylamine (mean concentration 3.1 nmol/l), N-nitrosodimethylamine (mean concentration 0.90 nmol/l) and N-nitrosopyrrolidine (mean concentration 0.38 nmol/l). Significant correlations between mean intragastric pH values and mean N-nitrosodi-n-butylamine level (P = 0.005) and total volatile N-nitrosamine contents (P = 0.009) were observed.
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Jugo Gástrico/química , Mucosa Gástrica/metabolismo , Enfermedades Gastrointestinales/metabolismo , Nitratos/metabolismo , Nitritos/metabolismo , Nitrosaminas/análisis , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Endoscopía Gastrointestinal , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Concentración de Iones de Hidrógeno , Masculino , Persona de Mediana Edad , Saliva/químicaRESUMEN
Formation of nitrite from ingested nitrate can result in several adverse health effects and implies a genotoxic risk as a consequence of endogenous formation of carcinogenic N-nitroso compounds. We studied the formation of volatile N-nitrosamines after intake of nitrate at the acceptable daily intake (ADI) level in combination with a fish meal rich in amines as nitrosatable precursors. Twenty-five volunteers consumed this meal during 7 consecutive days; a diet low in nitrate was consumed during 1 week before and 1 week after the test week. Nitrate intake at the ADI level resulted in a significant rise in mean salivary nitrate and nitrite concentrations. Mean urinary nitrate excretion increased from 76 mg/24 hr in the first control week to 194 and 165 mg/24 hr in the test week, followed by a decline to 77 mg/24 hr in the second control week. The urine samples were analyzed for volatile N-nitrosamines, and both N-nitrosodimethylamine (NDMA) and N-nitrosopiperidine (NPIP) were detected in the samples. Mean urinary NDMA excretion significantly increased from 287 ng/24 hr in the control week to 871 and 640 ng/24 hr in the test week and declined to 383 ng/24 hr in the second control week. Excretion of NPIP was not directly related to the nitrate intake and composition of the diet. Nitrate excretion and NDMA excretion were significantly correlated, as well as salivary nitrate and nitrite concentration and NDMA excretion. We conclude that nitrate intake at the ADI level in combination with a fish meal containing nitrosatable precursors increases NDMA excretion in urine and thus demonstrates increased formation of carcinogenic N-nitrosamines.
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Aminas/metabolismo , Dieta , Nitratos/metabolismo , Nitrosaminas/metabolismo , Compuestos de Potasio/metabolismo , Adolescente , Adulto , Femenino , Humanos , Persona de Mediana Edad , Países Bajos , Nitratos/efectos adversos , Nitrosaminas/orina , Compuestos de Potasio/efectos adversos , Saliva/metabolismo , Alimentos MarinosRESUMEN
It is known that lower-chlorinated biphenyls are metabolically activated to electrophilic quinoid species capable of binding to DNA. Also, certain metabolites are capable of redox cycling, thereby increasing oxidative stress in biological systems. In the present study, we tested mono-, di-, tri-, tetra-, penta-, hexa-, and heptachlorinated biphenyls for their ability to bind with DNA and to induce oxidative DNA damage. We present additional evidence that several PCB congeners form DNA adducts after metabolic activation, which can be detected by the nuclease P1- or butanol-enrichment procedures of the (32)P-postlabeling technique. Butanol and nuclease P1 enrichments showed different adduct recoveries, depending on the level of chlorination of the biphenyls. Application of the nuclease P1 enrichment showed that the incubation of 2-chloro-; 3, 4-dichloro-; 2,4,4'-trichloro-; 3,4,5-trichloro-; and 2,2',5, 5'-tetrachlorobiphenyl with calf thymus DNA and liver microsomes from rats treated with phenobarbital, followed by oxidation with a peroxidase, produced five to eight different DNA adducts. For these lower-chlorinated biphenyls, butanol enrichment generally showed a lower recovery. For some higher substituted congeners (3,3',4,4', 5-pentachloro-, 2,2',3,4,4',5'-hexachloro-, 2,2',4,4',5, 5'-hexachloro-, and 2,2',3,4,4',5,5'-heptachlorobiphenyl), after butanol enrichment a single dominant spot was observed, which was absent in the nuclease P1 procedure. After incubation of calf thymus DNA with either higher- or lower-chlorinated PCB congeners, we were not able to detect significantly increased levels of oxidative DNA damage above background levels, measured as 8-oxo-7, 8-dihydro-2'deoxyguanosine. In view of the carcinogenicity of PCB mixtures in animals and the ability of PCB metabolites to bind covalently to DNA, rats were orally treated with a mixture of PCBs (Aroclor 1242). PCB-DNA adduct levels were analyzed in PCB target organs: liver, thymus, glandular stomach, spleen, testes, seminal vesicles and prostate DNA. In vivo PCB-DNA adducts could not be detected by either the butanol- or by the NP1-enrichment procedure in rat target tissue DNA. Also, no differences in oxidative DNA damage could be observed between PCB-treated rats and controls. These results indicate a lack of DNA reactivity of PCB mixtures in vivo.
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Aductos de ADN/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Bifenilos Policlorados/toxicidad , Administración Oral , Animales , Peso Corporal/efectos de los fármacos , Marcaje Isotópico/métodos , Masculino , Microsomas Hepáticos/efectos de los fármacos , Radioisótopos de Fósforo , Bifenilos Policlorados/administración & dosificación , Próstata/efectos de los fármacos , Ratas , Ratas Endogámicas Lew , Endonucleasas Específicas del ADN y ARN con un Solo Filamento/química , Bazo/efectos de los fármacos , Testículo/efectos de los fármacos , Timo/efectos de los fármacos , Distribución Tisular , Pruebas de Toxicidad/métodosRESUMEN
To study the interaction of singlet oxygen (1O2) with DNA and the biological consequences of 1O2-induced DNA damage, we used the thermodissociable endoperoxide of 3,3'-(1,4 naphthalidene) dipropionate (NDPO2) as a generator of free 1O2 in reactions with (1) 2'-deoxynucleoside 3'-monophosphates (dNps), (2) an oligonucleotide (16-mer) having one deoxyguanine (dG), (3) native and denaturated rat kidney DNA and (4) single-stranded (ss) and double-stranded (ds) bacteriophage M13mp10 DNA. Using both anion exchange and reversed phase HPLC and 32P-postlabeling analyses, it was found that exposure of the various dNps to chemically generated 1O2 led to a detectable reaction with dGp and not with dAp, dCp, d5mCp or Tp. The reaction with dGp led to degradation of this nucleotide and the formation of a large number of reaction products, one of which could be identified as 7-hydro-8-oxo-2'-deoxyguanosine 3'-monophosphate (8-oxo-dGp). A second product could tentatively be identified as a formamido pyrimidine derivative of dGp (Fapy-dGp). When ss DNA, ds DNA or the oligonucleotide were exposed to 1O2, the formation of 8-oxo-dG could also be demonstrated. With the oligonucleotide, we found a so far unidentified reaction product. Under the same reaction conditions the yield of 8-oxo-dG was about 8-fold higher in ss DNA than in ds DNA. In ss DNA 8-oxo-dG seemed to be a more prominent product than in the case of reaction of 1O2 with free dGp. Reaction of 1O2 with ss or ds M13mp10 DNA led to biological inactivation of these DNAs, ss DNA being at least 100-fold more sensitive than ds DNA. It could be concluded that inactivation of the ss DNA must be largely due to 1O2-induced DNA lesions other than 8-oxo-dG. In agreement with the observed preferential reaction of 1O2 with dG most of the so far sequenced mutations, induced by 1O2 in a 144 bp mutation target sequence inserted in the lacZ alpha gene of ss or ds M13mp10 DNA, occurred at a G or G/C base pair respectively. A preference for G(C) to T(A) transversions can be observed for which 8-oxo-dG might have been responsible. In ss DNA a significant number of the mutations are characterized by the fact that a G is deleted.
Asunto(s)
Daño del ADN , ADN/química , ADN/metabolismo , Oligodesoxirribonucleótidos/química , Oxígeno , Animales , Bacteriófago M13/genética , Secuencia de Bases , ADN/efectos de la radiación , ADN de Cadena Simple/química , ADN de Cadena Simple/aislamiento & purificación , ADN Viral/química , ADN Viral/aislamiento & purificación , Escherichia coli/metabolismo , Indicadores y Reactivos , Riñón/metabolismo , Datos de Secuencia Molecular , Radioisótopos de Fósforo , Fotoquímica , Técnica de Dilución de Radioisótopos , Ratas , Oxígeno SingleteRESUMEN
BACKGROUND: In contrast to the postnatal period, little is known about telomere length (TL) during prenatal life. The decrease in placental TL remains unknown, although intra uterine growth retardation and preeclampsia are associated with shorter placental TL. The aim of this study is to assess the decrease of placental TL during the third trimester of gestation and to explore the role of potential "growth influencing factors". METHODS: The study sample consisted of 329 live-born twins from the East Flanders Prospective Twin Survey. TL was determined using a multiplex quantitative PCR method. Gestational age, sex, birth order, placental characteristics, parity, maternal and paternal age, diabetes, hypertension, smoking, alcohol use, and socio economic status (SES) were considered "growth influencing factors". Bivariable multilevel regression analysis with "growth influencing factors" was performed. RESULTS: Placental TL ranged from 4.3 kbp to 84.4 kbp with a median of 10.8 kbp. Ln(TL) decreased in a linear fashion with an estimated TL decreasing from 13.98 kbp at 28 weeks to 10.56 kbp at 42 weeks. The regression coefficient of gestational age became smaller if considered together with SES (b = -0.017; p = 0.08) or diabetes (b = -0.018; p = 0.07) and bigger if considered together with parity (b = -0.022; p = 0.02), indicating that part of the association between gestational age and telomere length is explained by these three confounding factors. CONCLUSION: Placental TL decreases during the third trimester of gestation of live-born twins with approximately 25% indicating that telomere shortening may play a role in aging of the placenta.
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Edad Gestacional , Paridad/fisiología , Placenta/metabolismo , Acortamiento del Telómero/fisiología , Telómero/metabolismo , Femenino , Humanos , Masculino , Embarazo , Tercer Trimestre del Embarazo , Estudios Prospectivos , GemelosAsunto(s)
Neoplasias Colorrectales/metabolismo , Pólipos Intestinales/metabolismo , Polienos/metabolismo , Adenocarcinoma/metabolismo , Estudios de Casos y Controles , Cromatografía Líquida de Alta Presión , Dieta , Grasas de la Dieta , Femenino , Humanos , Masculino , Análisis de Regresión , Factores SexualesAsunto(s)
Cromatografía de Gases y Espectrometría de Masas/métodos , Jugo Gástrico/química , Nitrosaminas/análisis , Nitrosaminas/orina , Determinación de la Acidez Gástrica , Humanos , Concentración de Iones de Hidrógeno , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Factores de TiempoRESUMEN
We studied the formation of carcinogenic nitrosamines during consumption of food rich in nitrate and amines, and its possible inhibition by use of an antibacterial mouthwash. Twelve volunteers were fed a diet containing the high-nitrate vegetables lettuce or spinach during two periods of four consecutive days, in combination with fish products containing high levels of amines as nitrosatable precursors. During the two periods, the subjects used an antibacterial mouthwash containing chlorhexidine or a control mouthwash without antibacterial activity. Twenty-four-hour urine samples were collected after consumption of the meals, and saliva samples were collected 1 h after each meal. The nitrate and nitrite contents of the urine and saliva samples were determined by spectrophotometry (for nitrite) and HPLC (for nitrate). The concentrations of volatile nitrosamines in the urine samples were determined by gas chromatography-mass spectrometry. Significant increases in mean urinary nitrate levels (from 59 to 135 mg/24 h) and in mean salivary nitrate levels (from 10 to 56 microg/ml) and salivary nitrite levels (from 2 to 11 microg/ml) were observed during the consumption of food rich in nitrate and amines, as well as a significant increase in the mean urinary excretion of total examined volatile nitrosamines (from 2 to 7 nmol/24 h) and of N-nitrosodimethylamine (from 1.2 to 2.9 nmol/24 h). Use of the antibacterial mouthwash resulted in a decrease in mean salivary nitrite levels from 16 to 3 microg/ml and a decrease in mean urinary excretion of N-nitrosomorpholine (from 7.0 to 0.3 nmol/24 h). For the whole data set, significant correlations were observed between nitrate intake in food and urinary nitrate (p = 0.01; r2 = 0.07) and between urinary nitrate and urinary N-nitrosodimethylamine (p = 0.002; r2 = 0.11). In conclusion, consumption of a diet rich in nitrate and amines increases the risk of formation of carcinogenic nitrosamines. Use of an antibacterial mouthwash containing chlorhexidine can result in inhibition of nitrosamine formation.
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Aminas/metabolismo , Ingestión de Alimentos , Antisépticos Bucales/farmacología , Nitratos/metabolismo , Nitrosaminas/metabolismo , Adulto , Animales , Clorhexidina/farmacología , Femenino , Humanos , Masculino , Nitratos/orina , Nitritos/metabolismo , Nitritos/orina , Nitrosaminas/orina , Saliva/química , Pastas de DientesRESUMEN
The fast atom bombardment collision-induced dissociation mass spectra of the [M-H]- ions of the 2-, 3-, 4- and 6-deoxy derivatives from methyl beta-D-galactopyranoside and some related compounds have been recorded. The fragmentation reactions of these quasimolecular ions and of OD-labelled analogues have been examined and related to the molecular structure. In some cases distinct and common mechanisms can be derived, but it is also clear from these experiments that not only the site of deprotonation of the molecules, but also the anticipated charge localization in the fragment ions strongly direct the fragmentation reactions.
Asunto(s)
Metilgalactósidos/química , Espectrometría de Masa Bombardeada por Átomos Veloces/métodosRESUMEN
Fecapentaene-12 (FP-12), a fecal unsaturated, ether-linked lipid excreted by most human individuals in Western populations, has been found to be a potent genotoxin in mammalian cells. Its mechanism of genotoxicity may be mediated by oxygen radical-induced DNA damage or by direct DNA alkylation, of which the relative importance remains to be determined. In the present study, induction of oxidative genetic damage by FP-12 has been investigated, in combination with the biological inactivation of single-stranded bacteriophage phi X-174 DNA. It was shown that formation of 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxodG), a marker for oxidative DNA damage, is induced dose dependently by FP-12 in 2'-deoxyguanosine (dG). It was demonstrated by application of radical scavengers that production of both the superoxide anion and singlet oxygen may be involved in the induction of 8-oxodG. The effect of OH radical scavenging appeared to be less pronounced. Enzymatic peroxidation of FP-12, which has been demonstrated to stimulate oxygen radical formation, was found to increase the hydroxylation ratio in dG, an effect which was less pronounced in single-stranded DNA and even absent in double-stranded DNA. No induction of 8-oxodG was observed after exposure of human skin fibroblasts to 60 microM FP-12 for 3 h in vitro. It was concluded that the induction of 8-oxodG by FP-12 is determined by the accessibility of the guanine molecule rather than the rate of oxygen radical formation. Although free radical formation is known to be stimulated by enzymatic peroxidation of FP-12, the inactivation of phi X-174 DNA spontaneously induced by FP-12 was found to be reduced by application of peroxidases. This furthermore demonstrates that the increased formation of reactive oxygen species by enzymatic peroxidation of FP-12 does not directly relate to increased induction of genotoxic effects. The fact that addition of radical scavengers shows limited effects on the inactivation of phi X-174 DNA suggests that the contribution of oxidative DNA damage to the genotoxic potential of FP-12 is only of minor importance.
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Daño del ADN , ADN/efectos de los fármacos , Mutágenos/toxicidad , Polienos/toxicidad , Animales , Bacteriófago phi X 174/genética , Óxidos N-Cíclicos/farmacología , ADN/metabolismo , ADN de Cadena Simple/efectos de los fármacos , ADN Viral/efectos de los fármacos , Radical Hidroxilo , Oxidación-Reducción , Ratas , Superóxido Dismutasa/farmacologíaRESUMEN
The DNA lesions induced by free 1O2 and the biological and mutagenic consequences of 1O2-induced DNA damage have been studied. Using anion exchange HPLC, reverse-phase HPLC with electrochemical detection and 32P-postlabelling methods, we have shown that 1O2 reacts with 2'-deoxyguanine 3'-monophosphate (dGp) but not with any other dNp. Reaction with dGp yields a large number of products; one minor product was identified as 7-hydro-8-oxo-2'-deoxyguanosine 3'-monophosphate (8-oxo-dGp), and a second tentatively as a formamidopyridine derivative of dGp. 8-Oxo-dGp was also found after reaction of 1O2 with single-stranded (ss) DNA, double-stranded (ds) DNA or an oligonucleotide (16-mer) having one G. With the oligonucleotide we found a second unidentified reaction product. With ss DNA, 8-oxo-dG was a much more prominent product than in the reaction of 1O2 with free dGp and the yield was about eight-fold higher than with ds DNA. This agrees with our finding that ss M13 DNA is at least 100-fold more sensitive than ds M13 DNA to biological inactivation by 1O2. The inactivation of ss M13 DNA must be largely due to 1O2-induced lesions other than 8-oxo-dG. In agreement with the observed preferential reaction of 1O2 with dG, most of the mutations induced by 1O2 in ss or ds M13mp10 DNA occurred at a G or G/C basepair, respectively. A preference for G(C) to T(A) transversions was observed for which 8-oxo-dG might have been responsible. In ss DNA, a significant number of mutations are characterized by the fact that a G is deleted.
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Daño del ADN , Oxígeno/toxicidad , Bacteriófago M13/efectos de los fármacos , Bacteriófago M13/genética , Secuencia de Bases , Cromatografía en Capa Delgada , ADN/análisis , ADN/efectos de los fármacos , ADN/genética , Análisis Mutacional de ADN , ADN Viral/efectos de los fármacos , ADN Viral/genética , Técnicas In Vitro , Datos de Secuencia Molecular , Radioisótopos de Fósforo , Oxígeno SingleteRESUMEN
BACKGROUND & AIMS: This study evaluated the effect of long-term gastric acid suppressive therapy with omeprazole on intragastric levels of carcinogenic N-nitrosamines and related parameters. METHODS: Forty-five patients on long-term omeprazole medication (mean, 35 months) and 13 healthy subjects without medication participated. Volatile N-nitrosamines were determined in gastric juice and urine. Intragastric pH, nitrite, nitrate, and H. pylori status were determined. DNA isolated from gastric biopsy specimens was analyzed for precarcinogenic alkyl-DNA adducts. RESULTS: The intragastric pH in patients was significantly higher compared with controls (P = 0.0001). Gastric nitrite levels in patients were nonsignificantly higher. There was no difference in total levels of intragastric volatile N-nitrosamines between patients and controls, however, urinary N-nitrosodimethylamine excretion was higher in patients (P = 0.001). On omeprazole, Helicobacter pylori-positive vs. -negative patients had a nonsignificantly higher intragastric nitrite level and higher urinary N-nitrosodimethylamine excretion. No alkyl-DNA adducts could be detected in gastric epithelium. CONCLUSIONS: Increased intragastric pH caused by long-term treatment with omeprazole does not result in increased intragastric levels of nitrite and volatile N-nitrosamines. The significantly higher urinary N-nitrosamine excretion implies the risk of increased endogenous formation of N-nitrosamines during long-term omeprazole treatment. This risk may be higher in H. pylori-positive patients.
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Antiulcerosos/efectos adversos , Infecciones por Helicobacter/patología , Helicobacter pylori/aislamiento & purificación , Concentración de Iones de Hidrógeno/efectos de los fármacos , Nitritos/análisis , Nitrosaminas/análisis , Omeprazol/efectos adversos , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Femenino , Jugo Gástrico/química , Jugo Gástrico/microbiología , Infecciones por Helicobacter/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Inhibidores de la Bomba de Protones , Factores de Riesgo , Neoplasias Gástricas/epidemiología , Neoplasias Gástricas/patologíaRESUMEN
We investigated the effects of smoking-induced oxidative stress in healthy volunteers (21 smokers versus 24 non-smokers) by quantifying various markers of oxidative DNA damage and repair, and antioxidative defense mechanisms. Lymphocytic 7-hydroxy-8-oxo-2'-deoxyguanosine (8-oxo-dG) levels measured by high performance liquid chromatography with electrochemical detection, were significantly lower in smokers as compared with non-smokers (38.6 +/- 5.2 versus 50.9 +/- 4.6/10(6) dG, P = 0.05). The levels of oxidized pyrimidine bases in lymphocytes of smokers quantified by the endonuclease III-modified comet assay were non-significantly lower than those of non-smokers (% DNA in tail: 13 +/- 3 versus 14 +/- 2; tail length: 69 +/- 13 versus 96 +/- 10; tail moment: 6416 +/- 1220 versus 7545 +/- 1234). Urinary excretion levels of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) assessed by enzyme-linked immunosorbent assay did not differ significantly between smokers and non-smokers (197 +/- 31 versus 240 +/- 33 ng/body mass index, P = 0.3). Overall DNA repair activity expressed as unscheduled DNA synthesis in blood leukocytes, was not significantly different between smokers and non-smokers (2.9 +/- 0.3 versus 3.3 +/- 0.3, P = 0.4). Plasma antioxidative capacity measured by the Trolox equivalent antioxidant capacity assay was slightly higher in smokers as compared with non-smokers (440 +/- 16 versus 400 +/- 15 microM Trolox equivalent, P = 0.09), and it was significantly related to lymphocytic 8-oxo-dG levels (r = 0.4, P = 0.001). Genotyping of human 8-OH-dG glycosylase/apurinic lyase and glutathione S-transferase M1 showed that a polymorphism in either or both of the two genes does not affect any of the quantified biomarkers. We conclude that oxidative stress imposed by cigarette smoking has a low impact upon certain pathways involved in DNA damage and the antioxidative defense system.
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Antioxidantes/metabolismo , Biomarcadores/sangre , Daño del ADN , Reparación del ADN , Desoxiguanosina/análogos & derivados , Estrés Oxidativo , 8-Hidroxi-2'-Desoxicoguanosina , Adulto , Cromatografía Líquida de Alta Presión , Desoxiguanosina/sangre , Desoxiguanosina/orina , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
In the multistage model of human colorectal tumorigenesis, both genetic and environmental factors play an important role. The identity of the environmental factors involved, however, still remains to be elucidated. As fecal bile acids are proposed as candidates, we compared the concentration of bile acids in fecal water from patients at different risk of developing colorectal cancer. In addition, pH of fecal water as well as its cytotoxicity to HT-29 colonic cells was determined. The high-risk group consisted of individuals diagnosed with one or more (tubulo)villous colorectal adenomas larger than 1 cm in diameter and containing moderate or severe dysplasia (N = 20). Subjects with colorectal adenomas smaller than 1 cm and showing only minor dysplasia were assigned to the medium risk group (N = 19). The control group consisted of persons with normal findings by colonoscopy (N = 25). The results show no significant differences in fecal water bile acid concentrations between the three groups. However, 46% of the observed cytotoxicity is explained in a regression model that includes pH and the concentrations of deoxycholic acid, cholic acid, and ursodeoxycholic acid. The pH of fecal water is found to be significantly lower in the high risk group as compared to the controls, suggesting that a relatively high fecal pH has a protective effect on the development of colorectal adenomas. Although hyperproliferation as a result of cytotoxicity has been suggested to contribute to tumor formation in the colon, the pH-dependent cytotoxicity of bile acids in fecal water was not found to be associated with adenoma formation in the present study.