117 results on '"Godschalk RW"'
Search Results
2. Transcriptional profiling of the acute pulmonary inflammatory response induced by LPS: role of neutrophils.
- Author
-
Güngör N, Pennings JL, Knaapen AM, Chiu RK, Peluso M, Godschalk RW, Van Schooten FJ, Güngör, Nejla, Pennings, Jeroen L A, Knaapen, Ad M, Chiu, Roland K, Peluso, Marco, Godschalk, Roger W L, and Van Schooten, Frederik J
- Abstract
Background: Lung cancer often develops in association with chronic pulmonary inflammatory diseases with an influx of neutrophils. More detailed information on inflammatory pathways and the role of neutrophils herein is a prerequisite for understanding the mechanism of inflammation associated cancer.Methods: In the present study, we used microarrays in order to obtain a global view of the transcriptional responses of the lung to LPS in mice, which mimics an acute lung inflammation. To investigate the influence of neutrophils in this process, we depleted mice from circulating neutrophils by treatment with anti-PMN antibodies prior to LPS exposure.Results: A total of 514 genes was greater than 1.5-fold differentially expressed in the LPS induced lung inflammation model. 394 of the 514 were up regulated genes mostly involved in cell cycle and immune/inflammation related processes, such as cytokine/chemokine activity and signalling. Down regulated genes represented nonimmune processes, such as development, metabolism and transport. Notably, the number of genes and pathways that were differentially expressed, was reduced when animals were depleted from circulating neutrophils, confirming the central role of neutrophils in the inflammatory response. Furthermore, there was a significant correlation between the differentially expressed gene list and the promutagenic DNA lesion M1dG, suggesting that it is the extent of the immune response which drives genetic instability in the inflamed lung. Several genes that were specifically regulated by the presence of activated neutrophils could be identified and these were mostly involved in interferon signalling, oxidative stress response and cell cycle progression. The latter possibly refers to a higher rate of cell turnover in the inflamed lung with neutrophils, suggesting that the neutrophil influx is associated with a higher risk for the accumulation and fixation of mutations.Conclusion: Gene expression profiling identified specific genes and pathways that are related to neutrophilic inflammation and could be associated to cancer development and indicate an active role of neutrophils in mediating the LPS induced inflammatory response in the mouse lung. [ABSTRACT FROM AUTHOR]- Published
- 2010
- Full Text
- View/download PDF
3. Maternal fatty acid status during pregnancy versus offspring inflammatory markers: a canonical correlation analysis of the MEFAB cohort.
- Author
-
Rouschop SH, Smolinska A, Gielen M, de Groot RHM, Zeegers MP, Opperhuizen A, van Schooten FJ, and Godschalk RW
- Abstract
The development of inflammatory lung disorders in children may be related to maternal fatty acid intake during pregnancy. We therefore examined maternal fatty acid (FA) status during pregnancy and its associations with inflammatory markers and lung conditions in the child by analyzing data from the MEFAB cohort using multivariate canonical correlation analysis (CCA). In the MEFAB cohort, 39 different phospholipid FAs were measured in maternal plasma at 16, 22 and 32 weeks of pregnancy, and at day of birth. Child inflammatory markers and self-reported doctor diagnosis of inflammatory lung disorders were assessed at 7 years of age. Using CCA, we found that maternal FA levels during pregnancy were significantly associated with child inflammatory markers at 7 years of age and that Mead acid (20:3n-9) was the most important FA for this correlation. To further verify the importance of Mead acid, we examined the relation between maternal Mead acid levels at the day of birth with the development of inflammatory lung disorders in children at age 7. After stratification for the child's sex, maternal Mead acid levels at day of birth were significantly related with self-reported doctor diagnosis of asthma and lung infections in boys, and bronchitis and total number of lung disorders in girls. Future studies should investigate whether the importance of Mead acid in the relation between maternal FA status and inflammation and lung disorders in the child is due to its role as biomarker for essential fatty acid deficiency or due to its own biological function as pro-inflammatory mediator., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Rouschop, Smolinska, Gielen, de Groot, Zeegers, Opperhuizen, van Schooten and Godschalk.)
- Published
- 2023
- Full Text
- View/download PDF
4. Perinatal High-Fat Diet Influences Ozone-Induced Responses on Pulmonary Oxidant Status and the Molecular Control of Mitophagy in Female Rat Offspring.
- Author
-
Rouschop SH, Snow SJ, Kodavanti UP, Drittij MJ, Maas LM, Opperhuizen A, van Schooten FJ, Remels AH, and Godschalk RW
- Subjects
- Animals, Animals, Newborn, Female, Gene Expression Profiling, Lung metabolism, Male, Maternal Nutritional Physiological Phenomena, Mitochondria metabolism, Oxidative Phosphorylation, Oxidative Stress, Pregnancy, Prenatal Exposure Delayed Effects etiology, Rats, Rats, Long-Evans, Diet, High-Fat adverse effects, Lung pathology, Mitochondria pathology, Mitophagy, Oxidants metabolism, Ozone adverse effects, Prenatal Exposure Delayed Effects pathology
- Abstract
Previous research has shown that a perinatal obesogenic, high-fat diet (HFD) is able to exacerbate ozone-induced adverse effects on lung function, injury, and inflammation in offspring, and it has been suggested that mitochondrial dysfunction is implicated herein. The aim of this study was to investigate whether a perinatal obesogenic HFD affects ozone-induced changes in offspring pulmonary oxidant status and the molecular control of mitochondrial function. For this purpose, female Long-Evans rats were fed a control diet or HFD before and during gestation, and during lactation, after which the offspring were acutely exposed to filtered air or ozone at a young-adult age (forty days). Directly following this exposure, the offspring lungs were examined for markers related to oxidative stress; oxidative phosphorylation; and mitochondrial fusion, fission, biogenesis, and mitophagy. Acute ozone exposure significantly increased pulmonary oxidant status and upregulated the molecular machinery that controls receptor-mediated mitophagy. In female offspring, a perinatal HFD exacerbated these responses, whereas in male offspring, responses were similar for both diet groups. The expression of the genes and proteins involved in oxidative phosphorylation and mitochondrial biogenesis, fusion, and fission was not affected by ozone exposure or perinatal HFD. These findings suggest that a perinatal HFD influences ozone-induced responses on pulmonary oxidant status and the molecular control of mitophagy in female rat offspring.
- Published
- 2021
- Full Text
- View/download PDF
5. Gene expression and DNA methylation as mechanisms of disturbed metabolism in offspring after exposure to a prenatal HF diet.
- Author
-
Rouschop SH, Karl T, Risch A, van Ewijk PA, Schrauwen-Hinderling VB, Opperhuizen A, van Schooten FJ, and Godschalk RW
- Subjects
- Animals, Cell Proliferation drug effects, Cell Proliferation genetics, DNA metabolism, DNA Methylation drug effects, DNA Methylation genetics, High-Throughput Nucleotide Sequencing, Liver drug effects, Liver metabolism, Male, Mice, Inbred C57BL, Oxidation-Reduction drug effects, Oxidative Stress drug effects, Phosphatidate Phosphatase genetics, Phosphatidate Phosphatase metabolism, Real-Time Polymerase Chain Reaction, Diet, High-Fat adverse effects, Lipid Metabolism drug effects, Lipid Metabolism genetics
- Abstract
Exposure to a prenatal high-fat (HF) diet leads to an impaired metabolic phenotype in mouse offspring. The underlying mechanisms, however, are not yet fully understood. Therefore, this study investigated whether the impaired metabolic phenotype may be mediated through altered hepatic DNA methylation and gene expression. We showed that exposure to a prenatal HF diet altered the offspring's hepatic gene expression of pathways involved in lipid synthesis and uptake (SREBP), oxidative stress response [nuclear factor (erythroid-derived 2)-like 2 (Nrf2)], and cell proliferation. The downregulation of the SREBP pathway related to previously reported decreased hepatic lipid uptake and postprandial hypertriglyceridemia in the offspring exposed to the prenatal HF diet. The upregulation of the Nrf2 pathway was associated with increased oxidative stress levels in offspring livers. The prenatal HF diet also induced hypermethylation of transcription factor (TF) binding sites upstream of lipin 1 ( Lpin1 ), a gene involved in lipid metabolism. Furthermore, DNA methylation of Lpin1 TF binding sites correlated with mRNA expression of Lpin1 These findings suggest that the effect of a prenatal HF diet on the adult offspring's metabolic phenotype are regulated by changes in hepatic gene expression and DNA methylation., (Copyright © 2019 Rouschop et al. Published by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2019
- Full Text
- View/download PDF
6. Polyunsaturated fatty acid status at birth, childhood growth, and cardiometabolic risk: a pooled analysis of the MEFAB and RHEA cohorts.
- Author
-
Stratakis N, Gielen M, Margetaki K, de Groot RHM, Apostolaki M, Chalkiadaki G, Vafeiadi M, Leventakou V, Karachaliou M, Godschalk RW, Kogevinas M, Stephanou EG, Zeegers MP, and Chatzi L
- Subjects
- Adult, Body Mass Index, Child, Child, Preschool, Diet, Female, Follow-Up Studies, Greece, Humans, Infant, Infant, Newborn, Maternal Exposure, Mothers, Netherlands, Risk, Waist Circumference physiology, Child Development physiology, Fatty Acids, Unsaturated blood, Pediatric Obesity epidemiology
- Abstract
Background/objectives: Polyunsaturated fatty acid (PUFA) status during pregnancy has been suggested to influence offspring obesity and cardiometabolic health. We assessed whether prenatal PUFA exposure is associated with rapid infant growth, childhood BMI, and cardiometabolic profile., Subjects/methods: In the Dutch MEFAB (n = 266) and Greek RHEA (n = 263) cohorts, we measured n-3 and n-6 PUFA concentrations in cord blood phospholipids, which reflect fetal exposure in late pregnancy. We defined rapid infant growth from birth to 6 months of age as an increase in weight z-score >0.67. We analyzed body mass index (BMI) as continuous and in categories of overweight/obesity at 4 and 6 years. We computed a cardiometabolic risk score at 6-7 years as the sum of waist circumference, non-high-density lipoprotein cholesterol and blood pressure z-scores. Associations of PUFAs with child health outcomes were assessed using generalized linear models for binary outcomes and linear regression models for continuous ones after adjusting for important covariates, and for the pooled estimates, a cohort indicator., Results: In pooled analyses, we found no association of PUFA levels with rapid infant growth, childhood BMI (β per SD increase in the total n-3:n-6 PUFA ratio = -0.04 SD; 99% CI: -0.15, 0.06; P = 0.65 at 4 years, and -0.05 SD; 99% CI: -0.18, 0.08; P = 0.78 at 6 years), and overweight/obesity. We also found no associations for clustered cardiometabolic risk and its individual components. The results were similar across cohorts., Conclusions: Our findings suggest that PUFA concentrations at birth are not associated with later obesity development and cardiometabolic risk in childhood.
- Published
- 2019
- Full Text
- View/download PDF
7. Bulky DNA adduct levels in normal-appearing colon mucosa, and the prevalence of colorectal adenomas.
- Author
-
Ho V, Peacock S, Massey TE, Godschalk RW, van Schooten FJ, Ashbury JE, Vanner SJ, and King WD
- Subjects
- Adenoma etiology, Adult, Female, Genetic Predisposition to Disease, Humans, Male, Middle Aged, Polymorphism, Genetic, Prevalence, Xeroderma Pigmentosum Group A Protein genetics, Colorectal Neoplasms etiology, DNA Adducts analysis, Intestinal Mucosa chemistry
- Abstract
Purpose: Examine the association between bulky DNA adduct levels in colon mucosa and colorectal adenoma prevalence, and explore the correlation between adduct levels in leukocytes and colon tissue., Methods: Bulky DNA adduct levels were measured using
32 P-postlabelling in biopsies of normal-appearing colon tissue and blood donated by 202 patients. Multivariable logistic regression was used to examine associations between DNA adducts, and interactions of DNA adduct-DNA repair polymorphisms, with the prevalence of colorectal adenomas. Correlation between blood and tissue levels of DNA adducts was evaluated using Spearman's correlation coefficient., Results: An interaction between bulky DNA adduct levels and XPA rs1800975 on prevalence of colorectal adenoma was observed. Among individuals with lower DNA repair activity, increased DNA adduct levels were associated with increased colorectal adenoma prevalence (OR = 1.41 per SD increase, 95%CI: 0.92-2.18). Conversely, among individuals with normal DNA activity, an inverse association was observed (OR = 0.60 per SD increase, 95%CI: 0.34-1.07). Blood and colon DNA adduct levels were inversely correlated (ρ = -0.20)., Conclusions: Among genetically susceptible individuals, higher bulky DNA adducts in the colon was associated with the prevalence of colorectal adenomas. The inverse correlation between blood and colon tissue measures demonstrates the importance of quantifying biomarkers in target tissues.- Published
- 2018
- Full Text
- View/download PDF
8. Sirtuin 1 genetic variation, energy balance and colorectal cancer risk by sex and subsite in the Netherlands Cohort Study.
- Author
-
Simons CCJM, Schouten LJ, Godschalk RW, van Schooten FJ, van den Brandt PA, and Weijenberg MP
- Subjects
- Aged, Body Mass Index, Cohort Studies, Female, Genetic Predisposition to Disease, Humans, Male, Middle Aged, Netherlands, Self Report, Sex Factors, Waist Circumference, Colorectal Neoplasms genetics, Polymorphism, Single Nucleotide, Sirtuin 1 genetics
- Abstract
Sirtuin 1 (SIRT1) is an energy-sensing protein, which may affect tumorigenesis. We used SIRT1 variants as time-independent indicators of SIRT1 involvement in carcinogenesis and we studied two tagging SIRT1 variants in relation to colorectal cancer (CRC) risk. We also evaluated known energy balance-related CRC risk factors within SIRT1 genotype strata. The Netherlands Cohort Study includes 120,852 individuals and has 20.3 years follow-up (case-cohort: n
subcohort = 5000; nCRC cases = 4667). At baseline, participants self-reported weight, weight at age 20, height, trouser/skirt size reflecting waist circumference, physical activity, and early life energy restriction. SIRT1 rs12778366 and rs10997870 were genotyped in toenail DNA available for ~75% of the cohort. Sex- and subsite-specific Cox hazard ratios (HRs) showed that the rs12778366 CC versus TT genotype decreased CRC and colon cancer risks in women (HRCRC = 0.53, 95% confidence interval: 0.30-0.94) but not men. Multiplicative interactions were observed between SIRT1 variants and energy balance-related factors in relation to CRC endpoints, but the direction of associations was not always conform expectation nor specific to one genotype stratum. In conclusion, these results support SIRT1 involvement in colon cancer development in women. No conclusions could be made regarding a modifying effect of SIRT1 variants on associations between energy balance-related factors and CRC risk.- Published
- 2018
- Full Text
- View/download PDF
9. PUFA status at birth and allergy-related phenotypes in childhood: a pooled analysis of the Maastricht Essential Fatty Acid Birth (MEFAB) and RHEA birth cohorts.
- Author
-
Stratakis N, Gielen M, Margetaki K, de Groot RHM, Apostolaki M, Chalkiadaki G, Vafeiadi M, Leventakou V, Godschalk RW, Kogevinas M, Stephanou EG, Zeegers MP, and Chatzi L
- Subjects
- Adult, Asthma epidemiology, Child, Cohort Studies, Eczema epidemiology, Fatty Acids, Omega-3 blood, Fatty Acids, Omega-6 blood, Female, Follow-Up Studies, Greece epidemiology, Humans, Infant, Newborn, Male, Netherlands epidemiology, Phenotype, Pregnancy, Respiratory Sounds, Risk, Fatty Acids, Essential, Fatty Acids, Unsaturated blood, Fetal Blood chemistry, Hypersensitivity blood, Prenatal Exposure Delayed Effects
- Abstract
Lower prenatal exposure to n-3 PUFA relative to n-6 PUFA has been hypothesised to influence allergy development, but evidence remains largely inconsistent. In the Dutch Maastricht Essential Fatty Acid Birth (MEFAB) (n 293) and Greek RHEA Mother-Child (n 213) cohorts, we investigated whether cord blood phospholipid PUFA concentrations are associated with symptoms of wheeze, asthma, rhinitis and eczema at the age of 6-7 years. Information on allergy-related phenotypes was collected using validated questionnaires. We estimated relative risks (RR) and 95 % CI for associations of PUFA with child outcomes using multivariable generalised linear regression models. In pooled analyses, higher concentration of the n-3 long-chain EPA and DHA and a higher total n-3:n-6 PUFA ratio were associated with lower risk of current wheeze (RR 0·61; 95 % CI 0·45, 0·82 per sd increase in EPA+DHA and 0·54; 95 % CI 0·39, 0·75 per unit increase in the n-3:n-6 ratio) and reduced asthma risk (RR 0·50; 95 % CI 0·31, 0·79 for EPA+DHA and 0·43; 95 % CI 0·26, 0·70 for the n-3:n-6 ratio). No associations were observed for other allergy-related phenotypes. The results were similar across cohorts. In conclusion, higher EPA and DHA concentrations and a higher n-3:n-6 fatty acid ratio at birth were associated with lower risk of child wheeze and asthma. Our findings suggest that dietary interventions resulting in a marked increase in the n-3:n-6 PUFA ratio, and mainly in n-3 long-chain PUFA intake in late gestation, may reduce the risk of asthma symptoms in mid-childhood.
- Published
- 2018
- Full Text
- View/download PDF
10. Altered gene expression profiles in the lungs of benzo[a]pyrene-exposed mice in the presence of lipopolysaccharide-induced pulmonary inflammation.
- Author
-
Shi Q, Fijten RR, Spina D, Riffo Vasquez Y, Arlt VM, Godschalk RW, and Van Schooten FJ
- Subjects
- Animals, Benzo(a)pyrene metabolism, DNA Adducts genetics, DNA Adducts metabolism, Disease Models, Animal, Gene Expression Profiling methods, Gene Regulatory Networks, Inflammation Mediators metabolism, Lung metabolism, Male, Metabolic Detoxication, Phase I, Metabolic Detoxication, Phase II, Mice, Inbred C57BL, Oligonucleotide Array Sequence Analysis, Pneumonia chemically induced, Pneumonia metabolism, Principal Component Analysis, Benzo(a)pyrene toxicity, Lipopolysaccharides, Lung drug effects, Pneumonia genetics, Transcriptome drug effects
- Abstract
Patients with inflammatory lung diseases are often additionally exposed to polycyclic aromatic hydrocarbons like B[a]P and B[a]P-induced alterations in gene expression in these patients may contribute to the development of lung cancer. Mice were intra-nasally treated with lipopolysaccharide (LPS, 20μg/mouse) to induce pulmonary inflammation and subsequently exposed to B[a]P (0.5mg/mouse) by intratracheal instillation. Gene expression changes were analyzed in mouse lungs by RNA microarrays. Analysis of genes that are known to be involved in the cellular response to B[a]P indicated that LPS significantly inhibited gene expression of various enzymes linked to B[a]P metabolism, which was confirmed by phenotypic analyses of enzyme activity. Ultimately, these changes resulted in higher levels of B[a]P-DNA adducts in the lungs of mice exposed to B[a]P with prior LPS treatment compared to the lungs of mice exposed to B[a]P alone. Using principle component analysis (PCA), we found that of all the genes that were significantly altered in their expression, those that were able to separate the different exposure conditions were predominantly related to immune-response. Moreover, an overall analysis of differentially expressed genes indicated that cell-cell adhesion and cell-cell communication was inhibited in lungs of mice that received both B[a]P and LPS. Our results indicate that pulmonary inflammation increased the genotoxicity of B[a]P via inhibition of both phase I and II metabolism. Therefore, inflammation could be a critical contributor to B[a]P-induced carcinogenesis in humans., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
11. Polyunsaturated fatty acid levels at birth and child-to-adult growth: Results from the MEFAB cohort.
- Author
-
Stratakis N, Gielen M, Margetaki K, Godschalk RW, van der Wurff I, Rouschop S, Ibrahim A, Antoniou E, Chatzi L, de Groot RHM, and Zeegers MP
- Subjects
- Adiposity, Adolescent, Adult, Body Height drug effects, Body Mass Index, Child, Child, Preschool, Female, Fetal Blood metabolism, Humans, Infant, Male, Obesity blood, Obesity pathology, Parturition, Pregnancy, Prenatal Exposure Delayed Effects, Young Adult, Fatty Acids, Omega-3 blood, Fatty Acids, Unsaturated blood, Obesity metabolism, Phospholipids blood
- Abstract
Background: Prenatal exposure to polyunsaturated fatty acids (PUFAs) may influence childhood growth. However, available evidence mostly derived from short-term studies is inconsistent., Objective: To assess whether fetal PUFA exposure is associated with height and body mass index (BMI), a common measure of adiposity, from 6 months to 23 years of age., Methods: In the MEFAB cohort, we assessed cord blood phospholipid n-3 and n-6 PUFA levels, reflecting fetal exposure in late pregnancy. For 250 (45.2% females) participants, we collected a total of 1770 (n= 802 for females) repeated growth measurements from infancy to young adulthood. We examined sex-specific associations of PUFAs with height and BMI at different developmental ages (infant: 6 months; toddler: 2 years; pre-schooler: 4 years; school-aged child: 7 years; adolescent: 12 years; and young adult: 23 years) using fractional polynomial mixed models adjusted for important covariates., Results: Higher n-3 PUFA levels were associated with higher infant length in males (β= 0.44cm [95% CI: 0.07, 0.82] per SD increase), whereas, for females, higher n-6 PUFA concentrations were associated with lower length in infancy (β= -0.69cm [95% CI: -1.08, -0.30] per SD increase). A higher ratio of n-3 to n-6 PUFAs was associated with higher infant length in both sexes (β= 0.40cm [95% CI: 0.01, 0.78] and 0.42cm [95% CI: 0.05, 0.79] per unit increase for males and females, respectively). These associations were not detectable later in childhood and young adulthood. No associations with BMI were found at any time point examined., Conclusions: Our findings suggest a small sex-specific influence of PUFA status at birth on length in infancy, but this does not persist in later life up to young adulthood. PUFA status at birth does not seem to affect BMI from infancy till young adulthood., (Copyright © 2017 Elsevier Ltd. All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
12. Acidic cellular microenvironment modifies carcinogen-induced DNA damage and repair.
- Author
-
Shi Q, Maas L, Veith C, Van Schooten FJ, and Godschalk RW
- Subjects
- A549 Cells, Benzo(a)pyrene metabolism, Carcinogens metabolism, Cell Culture Techniques, Cytochrome P-450 CYP1A1 metabolism, Cytochrome P-450 CYP1B1 metabolism, Humans, Hydrogen-Ion Concentration, Metabolic Networks and Pathways, Benzo(a)pyrene toxicity, Carcinogens toxicity, Cellular Microenvironment drug effects, DNA Damage, DNA Repair
- Abstract
Chronic inflammation creates an acidic microenvironment, which plays an important role in cancer development. To investigate how low pH changes the cellular response to the carcinogen benzo[a]pyrene (B[a]P), we incubated human pulmonary epithelial cells (A549 and BEAS-2B) with nontoxic doses of B[a]P using culturing media of various pH's (extracellular pH (pH
e ) of 7.8, 7.0, 6.5, 6.0 and 5.5) for 6, 24 and 48 h. In most incubations (pHe 7.0-6.5), the pH in the medium returned to the physiological pH 7.8 after 48 h, but at the lowest pH (pHe < 6.0), this recovery was incomplete. Similar changes were observed for the intracellular pH (pHi ). We observed that acidic conditions delayed B[a]P metabolism and at t = 48 h, and the concentration of unmetabolized extracellular B[a]P and B[a]P-7,8-diol was significantly higher in acidic samples than under normal physiological conditions (pHe 7.8) for both cell lines. Cytochrome P450 (CYP1A1/CYP1B1) expression and its activity (ethoxyresorufin-O-deethylase activity) were repressed at low pHe after 6 and 24 h, but were significantly higher at t = 48 h. In addition, a DNA repair assay showed that the incision activity was ~80% inhibited for 6 h at low pHe and concomitant exposure to B[a]P. However, at t = 48 h, the incision activity recovered to more than 100% of the initial activity observed at neutral pHe . After 48 h, higher B[a]P-DNA adduct levels and γ-H2AX foci were observed at low pH samples than at pHe 7.8. In conclusion, acidic pH delayed the metabolism of B[a]P and inhibited DNA repair, ultimately leading to increased B[a]P-induced DNA damage.- Published
- 2017
- Full Text
- View/download PDF
13. Effect of interleukin (IL)-8 on benzo[a]pyrene metabolism and DNA damage in human lung epithelial cells.
- Author
-
Shi Q, Boots AW, Maas L, Veith C, van Kuijk K, Haenen GR, Godschalk RW, and Van Schooten FJ
- Subjects
- Carcinogens toxicity, Cell Line, Cell Survival drug effects, Cytochrome P-450 CYP1A1 genetics, Cytochrome P-450 CYP1A1 metabolism, Cytochrome P-450 CYP1B1 genetics, Cytochrome P-450 CYP1B1 metabolism, Epithelial Cells metabolism, Humans, Lung cytology, NADP metabolism, NADPH Oxidases genetics, NADPH Oxidases metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, Benzo(a)pyrene toxicity, DNA Damage drug effects, Epithelial Cells drug effects, Interleukin-8 pharmacology
- Abstract
It has been well established that inflammation and concurrent mutagenic exposures drive the carcinogenic process in a synergistic way. To elucidate the role of the inflammatory cytokine IL-8 in this process, we studied its effect on the activation and deactivation of the chemical mutagen benzo[a]pyrene B[a]P in the immortalized pulmonary BEAS-2B cell line. After 24h incubation with B[a]P in the presence or absence of IL-8, the B[a]P induced cytochrome P450 1A1 and 1B1 (CYP1A1 and CYP1B1) gene expression and CYP1A1 enzyme activity was significantly higher in the presence of the cytokine. Consistent with these findings, we observed higher concentration of the metabolite B[a]P-7,8-diol under concurrent IL-8 treatment conditions. Interestingly, we also found higher concentrations of unmetabolized B[a]P. To explain this, we examined the downstream effects of IL-8 on NADPH oxidases (NOXes). IL-8 lowered the intracellular NADPH level, but this effect could not explain the changes in B[a]P metabolism. IL-8 also significantly depleted intracellular glutathione (GSH), which also resulted in enhanced levels of unmetabolized B[a]P, but increased concentrations of the metabolite B[a]P-7,8-diol. No differences in B[a]P-DNA adducts level were found between B[a]P and B[a]P combined with IL-8, and this might be due to a 3-fold increase in nucleotide excision repair (NER) after IL-8 treatment. These findings suggest that IL-8 increased the formation of B[a]P-7,8-diol despite an overall delayed B[a]P metabolism via depletion of GSH, but DNA damage levels were unaffected due to an increase in NER capacity., (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
14. The Ageing Brain: Effects on DNA Repair and DNA Methylation in Mice.
- Author
-
Langie SA, Cameron KM, Ficz G, Oxley D, Tomaszewski B, Gorniak JP, Maas LM, Godschalk RW, van Schooten FJ, Reik W, von Zglinicki T, and Mathers JC
- Abstract
Base excision repair (BER) may become less effective with ageing resulting in accumulation of DNA lesions, genome instability and altered gene expression that contribute to age-related degenerative diseases. The brain is particularly vulnerable to the accumulation of DNA lesions; hence, proper functioning of DNA repair mechanisms is important for neuronal survival. Although the mechanism of age-related decline in DNA repair capacity is unknown, growing evidence suggests that epigenetic events (e.g., DNA methylation) contribute to the ageing process and may be functionally important through the regulation of the expression of DNA repair genes. We hypothesize that epigenetic mechanisms are involved in mediating the age-related decline in BER in the brain. Brains from male mice were isolated at 3-32 months of age. Pyrosequencing analyses revealed significantly increased Ogg1 methylation with ageing, which correlated inversely with Ogg1 expression. The reduced Ogg1 expression correlated with enhanced expression of methyl-CpG binding protein 2 and ten-eleven translocation enzyme 2. A significant inverse correlation between Neil1 methylation at CpG-site2 and expression was also observed. BER activity was significantly reduced and associated with increased 8-oxo-7,8-dihydro-2'-deoxyguanosine levels. These data indicate that Ogg1 and Neil1 expression can be epigenetically regulated, which may mediate the effects of ageing on DNA repair in the brain.
- Published
- 2017
- Full Text
- View/download PDF
15. A systematic SNP selection approach to identify mechanisms underlying disease aetiology: linking height to post-menopausal breast and colorectal cancer risk.
- Author
-
Elands RJ, Simons CC, Riemenschneider M, Isaacs A, Schouten LJ, Verhage BA, Van Steen K, Godschalk RW, van den Brandt PA, Stoll M, and Weijenberg MP
- Subjects
- Breast Neoplasms etiology, Colorectal Neoplasms etiology, Female, Genetic Predisposition to Disease, Genome-Wide Association Study, Humans, Body Height genetics, Breast Neoplasms genetics, Colorectal Neoplasms genetics, Polymorphism, Single Nucleotide, Postmenopause
- Abstract
Data from GWAS suggest that SNPs associated with complex diseases or traits tend to co-segregate in regions of low recombination, harbouring functionally linked gene clusters. This phenomenon allows for selecting a limited number of SNPs from GWAS repositories for large-scale studies investigating shared mechanisms between diseases. For example, we were interested in shared mechanisms between adult-attained height and post-menopausal breast cancer (BC) and colorectal cancer (CRC) risk, because height is a risk factor for these cancers, though likely not a causal factor. Using SNPs from public GWAS repositories at p-values < 1 × 10
-5 and a genomic sliding window of 1 mega base pair, we identified SNP clusters including at least one SNP associated with height and one SNP associated with either post-menopausal BC or CRC risk (or both). SNPs were annotated to genes using HapMap and GRAIL and analysed for significantly overrepresented pathways using ConsensuspathDB. Twelve clusters including 56 SNPs annotated to 26 genes were prioritised because these included at least one height- and one BC risk- or CRC risk-associated SNP annotated to the same gene. Annotated genes were involved in Indian hedgehog signalling (p-value = 7.78 × 10-7 ) and several cancer site-specific pathways. This systematic approach identified a limited number of clustered SNPs, which pinpoint potential shared mechanisms linking together the complex phenotypes height, post-menopausal BC and CRC.- Published
- 2017
- Full Text
- View/download PDF
16. Energy restriction at young age, genetic variants in the insulin-like growth factor pathway and colorectal cancer risk in the Netherlands Cohort Study.
- Author
-
Simons CC, Schouten LJ, Godschalk RW, van Engeland M, van den Brandt PA, van Schooten FJ, and Weijenberg MP
- Subjects
- Alleles, Cohort Studies, Female, Genotype, Humans, Male, Middle Aged, Netherlands, Risk Factors, Signal Transduction genetics, Colorectal Neoplasms etiology, Colorectal Neoplasms genetics, Genetic Predisposition to Disease genetics, Insulin-Like Growth Factor I genetics, Polymorphism, Single Nucleotide genetics
- Abstract
The energy restriction (ER)-colorectal cancer (CRC) association is inconsistent in literature. To strengthen the biological plausibility of the ER-CRC association, we investigated whether genetic variation in the insulin-like growth factor (IGF) pathway, a putative underlying mechanism, modulated this association in the Netherlands Cohort Study. Participants completed a questionnaire (n = 120,852) and provided toenail clippings for DNA (∼75%) at baseline. Individuals living in a Western city during the Hunger Winter (1944-45) or Western rural versus non-Western area were exposed to (severe) ER at young age. Genotyping was performed for 3,768 subcohort members and 2,580 CRC cases (case-cohort with 16.3 years follow-up). Cox hazard ratios for CRC were estimated across combined categories of ER and a genetic sum score of unfavorable alleles based on 18 single nucleotide polymorphisms in IGF-related genes and ER and an IGF1 19-CA repeat polymorphism. The reference included ER exposed individuals, so that increased hazard ratios were expected in higher combined categories for calculating relative excess risks due to interaction (additive interactions). Wald tests for multiplicative interactions were also performed. Multiplicative and additive interactions were nonsignificant. Combined ER-genetic sum score categories showed increasing CRC risks in men, but confidence intervals were wide. Women carrying two variant IGF1 19-CA repeat alleles versus those carrying two wild type IGF1 19-CA repeat alleles were at an ∼50% decreased CRC risk, irrespective of ER exposure. In conclusion, data indicate that the IGF pathway might be involved in the ER-CRC association in men, but not women, although interactions were nonsignificant, hampering definite conclusions., (© 2016 UICC.)
- Published
- 2017
- Full Text
- View/download PDF
17. The influence of single nucleotide polymorphisms on the association between dietary acrylamide intake and endometrial cancer risk.
- Author
-
Hogervorst JG, van den Brandt PA, Godschalk RW, van Schooten FJ, and Schouten LJ
- Subjects
- Acrylamide administration & dosage, Acrylamide metabolism, Aged, Carcinogens administration & dosage, Carcinogens metabolism, Carcinogens toxicity, Cohort Studies, Cytochrome P-450 CYP2E1 genetics, Endometrial Neoplasms enzymology, Female, Gene Deletion, Genetic Association Studies, Glutathione Transferase genetics, Humans, Middle Aged, Proportional Hazards Models, Prospective Studies, Risk Assessment, Risk Factors, Acrylamide toxicity, Diet adverse effects, Endometrial Neoplasms etiology, Endometrial Neoplasms genetics, Polymorphism, Single Nucleotide
- Abstract
It is unclear whether the association between dietary acrylamide intake and endometrial cancer risk as observed in some epidemiological studies reflects a causal relationship. We aimed at clarifying the causality by analyzing acrylamide-gene interactions for endometrial cancer risk. The prospective Netherlands Cohort Study on diet and cancer includes 62,573 women, aged 55-69 years. At baseline, a random subcohort of 2589 women was selected for a case cohort analysis approach. Acrylamide intake of subcohort members and endometrial cancer cases (n = 315) was assessed with a food frequency questionnaire. Single nucleotide polymorphisms (SNPs) in genes in acrylamide metabolism, sex steroid systems, oxidative stress and DNA repair were assessed through a MassARRAY iPLEX Platform. Interaction between acrylamide and SNPs was assessed with Cox proportional hazards analysis, based on 11.3 years of follow-up. Among the results for 57 SNPs and 2 gene deletions, there were no statistically significant interactions after adjustment for multiple testing. However, there were nominally statistically significant interactions for SNPs in acrylamide-metabolizing enzymes: CYP2E1 (rs915906 and rs2480258) and the deletions of GSTM1 and GSTT1. Although in need of confirmation, the interactions between acrylamide intake and CYP2E1 SNPs contribute to the evidence for a causal relationship between acrylamide and endometrial cancer risk., Competing Interests: Dr. Leo Schouten received compensation as a member of a scientific advisory panel on acrylamide risk assessment of the European Food Safety Authority. The other authors have no potential competing financial interests to declare.
- Published
- 2016
- Full Text
- View/download PDF
18. Potential role of gene-environment interactions in ion transport mechanisms in the etiology of renal cell cancer.
- Author
-
Deckers IA, van den Brandt PA, van Engeland M, van Schooten FJ, Godschalk RW, Keszei AP, Hogervorst JG, and Schouten LJ
- Abstract
We investigated the ion transport mechanism (ITM) in renal cell cancer (RCC) etiology using gene-environment interactions between candidate single nucleotide polymorphisms (SNPs) and associated environmental factors, including dietary intakes of sodium, potassium and fluid, hypertension and diuretic medication. A literature-based selection of 13 SNPs in ten ITM genes were successfully genotyped in toenail DNA of 3,048 subcohort members and 419 RCC cases from the Netherlands Cohort Study. Diet and lifestyle were measured with baseline questionnaires. Cox regression analyses were conducted for main effects and gene-environment interactions. ADD1_rs4961 was significantly associated with RCC risk, showing a Hazard Ratio (HR) of 1.24 (95% confidence intervals (CI): 1.01-1.53) for the GT + TT (versus GG) genotype. Four of 65 tested gene-environment interactions were statistically significant. Three of these interactions clustered in SLC9A3_rs4957061, including the ones with fluid and potassium intake, and diuretic medication. For fluid intake, the RCC risk was significantly lower for high versus low intake in participants with the CC genotype (HR(95% CI): 0.47(0.26-0.86)), but not for the CT + TT genotype (P-interaction = 0.002). None of the main genetic effects and gene-environment interactions remained significant after adjustment for multiple testing. Data do not support the general hypothesis that the ITM is a disease mechanism in RCC etiology.
- Published
- 2016
- Full Text
- View/download PDF
19. Does Ataxia Telangiectasia Mutated (ATM) protect testicular and germ cell DNA integrity by regulating the redox status?
- Author
-
Godschalk RW, Vanhees K, Maas L, Drittij MJ, Pachen D, van Doorn-Khosrovani Sv, van Schooten FJ, and Haenen GR
- Subjects
- 8-Hydroxy-2'-Deoxyguanosine, Animals, Ataxia Telangiectasia Mutated Proteins genetics, Comet Assay, DNA Damage, Deoxyguanosine analogs & derivatives, Deoxyguanosine metabolism, Genotype, Glucosephosphate Dehydrogenase metabolism, Male, Mice, Mutation, Oxidation-Reduction, Oxidative Stress, Glutathione metabolism, Spermatozoa metabolism, Testis metabolism
- Abstract
A balanced redox homeostasis in the testis is essential for genetic integrity of sperm. Reactive oxygen species can disturb this balance by oxidation of glutathione, which is regenerated using NADPH, formed by glucose-6-phosphate dehydrogenase (G6PDH). G6PDH is regulated by the Ataxia Telangiectasia Mutated (Atm) protein. Therefore, we studied the redox status and DNA damage in testes and sperm of mice that carried a deletion in Atm. The redox status in heterozygote mice, reflected by glutathione levels and antioxidant capacity, was lower than in wild type mice, and in homozygotes the redox status was even lower. The redox status correlated with oxidative DNA damage that was highest in mice that carried Atm deletions. Surprisingly, G6PDH activity was highest in homozygotes carrying the deletion. These data indicate that defective Atm reduces the redox homeostasis of the testis and genetic integrity of sperm by regulating glutathione levels independently from G6PDH activity., (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
20. Association between prenatal and current exposure to selected LCPUFAs and school performance at age 7.
- Author
-
van der Wurff IS, Bakker EC, Hornstra G, Kirschner PA, Gielen M, Godschalk RW, Kremers S, Zeegers MP, and de Groot RH
- Subjects
- Adult, Arachidonic Acid blood, Child, Child Development physiology, Educational Status, Eicosapentaenoic Acid blood, Female, Humans, Male, Pregnancy, Prenatal Care, Cognition physiology, Docosahexaenoic Acids blood, Fatty Acids, Unsaturated blood, Umbilical Cord metabolism
- Abstract
Introduction: Long-chain polyunsaturated fatty acids (LCPUFAs) are important for brain functioning and might, thus, influence cognition and school performance. However, research investigating LCPUFAs relationships with school performance is limited. The objective of this study was to determine the association between levels of the LCPUFAs docosahexaenoic acid (DHA), arachidonic acid (AA), eicosapentaenoic acid (EPA) and n-6 docosapentaenoic acid (Osbond acid, ObA) at study entry, 22 weeks of pregnancy, 32 weeks of pregnancy, at partus, in umbilical cord plasma and child's plasma at age 7 and school performance scores at age 7., Methods: Data from the Maastricht Essential Fatty Acid Birth cohort (MEFAB) were used for this study. Fatty acid levels of plasma phospholipids were measured in maternal blood plasma at study entry, 22 weeks of pregnancy, 32 weeks of pregnancy and partus. Childs fatty acid levels of plasma phospholipids were measured a in umbilical cord blood plasma, and in blood plasma of the child at age 7. Scores on national standardised tests for spelling, reading and arithmetic at age 7 were obtained via the school (scores were available for 149, 159 and 155 children, respectively). Associations between LCPUFA levels and school performance scores were analysed with categorical regression analyses with correction for covariates (smoking, maternal education, sex, breastfeeding, maternal intelligence, birth weight and BMI at age 7)., Results: Significant (p<0.001) associations between DHA level at age 7 and both reading (β=0.158) and spelling (β=0.146) were found. Consistent significant negative associations were observed between all maternal DHA plasma levels and arithmetic scores at age 7 (all p<0.001, all β<-0.019). Additional significant negative associations were observed between maternal LCPUFA plasma levels at study entry and both reading and spelling scores at age 7; these associations were less consistent., Conclusion: Plasma DHA levels at age 7 were positively associated with reading and spelling scores at age 7. Consistent significant negative associations between maternal plasma DHA levels and arithmetic scores of the child at age 7 were found. Although this is an observational study, which cannot proof causality, the consistent negative associations observed between maternal plasma DHA levels and the arithmetic scores of the children at age 7 calls upon prudence when considering DHA supplementation during pregnancy., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
21. The impact of p53 on DNA damage and metabolic activation of the environmental carcinogen benzo[a]pyrene: effects in Trp53(+/+), Trp53(+/-) and Trp53(-/-) mice.
- Author
-
Krais AM, Speksnijder EN, Melis JP, Indra R, Moserova M, Godschalk RW, van Schooten FJ, Seidel A, Kopka K, Schmeiser HH, Stiborova M, Phillips DH, Luijten M, and Arlt VM
- Subjects
- Activation, Metabolic, Animals, Benzo(a)pyrene metabolism, Carcinogens, Environmental metabolism, Cytochrome P-450 CYP1A1 metabolism, DNA Adducts metabolism, DNA Damage drug effects, Inactivation, Metabolic, Kidney drug effects, Kidney metabolism, Male, Mice, Inbred C57BL, Mice, Mutant Strains, Microsomes, Liver drug effects, Microsomes, Liver metabolism, NAD(P)H Dehydrogenase (Quinone) metabolism, Tumor Suppressor Protein p53 metabolism, Benzo(a)pyrene pharmacokinetics, Carcinogens, Environmental pharmacokinetics, DNA Damage genetics, Tumor Suppressor Protein p53 genetics
- Abstract
The tumour suppressor p53 is one of the most important cancer genes. Previous findings have shown that p53 expression can influence DNA adduct formation of the environmental carcinogen benzo[a]pyrene (BaP) in human cells, indicating a role for p53 in the cytochrome P450 (CYP) 1A1-mediated biotransformation of BaP in vitro. We investigated the potential role of p53 in xenobiotic metabolism in vivo by treating Trp53(+/+), Trp53(+/-) and Trp53(-/-) mice with BaP. BaP-DNA adduct levels, as measured by (32)P-postlabelling analysis, were significantly higher in liver and kidney of Trp53(-/-) mice than of Trp53(+/+) mice. Complementarily, significantly higher amounts of BaP metabolites were also formed ex vivo in hepatic microsomes from BaP-pretreated Trp53(-/-) mice. Bypass of the need for metabolic activation by treating mice with BaP-7,8-dihydrodiol-9,10-epoxide resulted in similar adduct levels in liver and kidney in all mouse lines, confirming that the influence of p53 is on the biotransformation of the parent compound. Higher BaP-DNA adduct levels in the livers of Trp53(-/-) mice correlated with higher CYP1A protein levels and increased CYP1A enzyme activity in these animals. Our study demonstrates a role for p53 in the metabolism of BaP in vivo, confirming previous in vitro results on a novel role for p53 in CYP1A1-mediated BaP metabolism. However, our results also suggest that the mechanisms involved in the altered expression and activity of the CYP1A1 enzyme by p53 in vitro and in vivo are different.
- Published
- 2016
- Full Text
- View/download PDF
22. Erratum: "Environmental, Dietary, Maternal, and Fetal Predictors of Bulky DNA Adducts in Cord Blood: A European Mother-Child Study (NewGeneris)".
- Author
-
Pedersen M, Mendez MA, Schoket B, Godschalk RW, Espinosa A, Landström A, Villanueva CM, Merlo DF, Fthenou E, Gracia-Lavedan E, van Schooten FJ, Hoek G, Brunborg G, Meltzer HM, Alexander J, Nielsen JK, Sunyer J, Wright J, Kovács K, de Hoogh K, Gutzkow KB, Hardie LJ, Chatzi L, Knudsen LE, Anna L, Ketzel M, Haugen M, Botsivali M, Nieuwenhuijsen MJ, Cirach M, Toledano MB, Smith RB, Fleming S, Agramunt S, Kyrtopoulos SA, Lukács V, Kleinjans JC, Segerbäck D, and Kogevinas M
- Published
- 2016
- Full Text
- View/download PDF
23. The oxidation of p-phenylenediamine, an ingredient used for permanent hair dyeing purposes, leads to the formation of hydroxyl radicals: Oxidative stress and DNA damage in human immortalized keratinocytes.
- Author
-
Zanoni TB, Hudari F, Munnia A, Peluso M, Godschalk RW, Zanoni MV, den Hartog GJ, Bast A, Barros SB, Maria-Engler SS, Hageman GJ, and de Oliveira DP
- Subjects
- Cell Line, Chromatography, Liquid, DNA Damage drug effects, Dose-Response Relationship, Drug, Humans, Hydrogen Peroxide metabolism, Keratinocytes metabolism, Malondialdehyde metabolism, Reactive Oxygen Species metabolism, Skin cytology, Tandem Mass Spectrometry, Hair Dyes analysis, Hydroxyl Radical metabolism, Keratinocytes drug effects, Oxidative Stress drug effects, Phenylenediamines toxicity
- Abstract
The hair-dyeing ingredient, p-phenylenediamine (PPD), was previously reported to be mutagenic, possibly by inducing oxidative stress. However, the exact mechanism of PPD in inducing oxidative stress upon skin exposure during hair-dyeing in human keratinocytes remains unknown. The aim of our studies was therefore to investigate the toxicity of PPD and its by-products in human immortalized keratinocytes (HaCaT) after auto-oxidation and after reaction with hydrogen peroxide (H2O2). We found that the PPD half maximal effective cytotoxic concentration (EC50) to HaCaT is 39.37 and 35.63 μg/mL after 24 and 48 h, respectively, without addition of H2O2 to induce oxidation. When PPD (10 or 100 μg/mL) is combined with 10.5 μg/mL of H2O2, intracellular ROS production by HaCaT after 1 h was significantly increased and enhanced levels of DNA damage were observed after 4 h of exposure. After 24 h incubations, 20 μg/mL of PPD increased the level of DNA oxidation in HaCaT. Also, we found that the in vitro reaction between PPD and H2O2, even below the maximum allowance by cosmetic industries, released hydroxyl radicals which can damage DNA. Taken together, we conclude that PPD alone and when combined with H2O2 increases the formation of reactive oxygen species in human keratinocytes, leading to oxidative stress and subsequent DNA damage. These alterations suggest that the mechanism by which PPD exposure, alone or combined with H2O2, damages keratinocytes by the formation of the high reactive HO∙ radicals., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
24. Genetic Variants in the Insulin-like Growth Factor Pathway and Colorectal Cancer Risk in the Netherlands Cohort Study.
- Author
-
Simons CC, Schouten LJ, Godschalk RW, van Engeland M, van den Brandt PA, van Schooten FJ, and Weijenberg MP
- Subjects
- Aged, Alleles, Colorectal Neoplasms metabolism, Female, Gene Frequency, Genetic Association Studies, Genotype, Humans, Male, Middle Aged, Netherlands epidemiology, Polymorphism, Single Nucleotide, Proportional Hazards Models, Risk, Signal Transduction, Somatomedins metabolism, Colorectal Neoplasms epidemiology, Colorectal Neoplasms genetics, Genetic Predisposition to Disease, Genetic Variation, Somatomedins genetics
- Abstract
Interrelationships between insulin-like growth factors (IGFs), hyperinsulinaemia, diabetes, and colorectal cancer (CRC) indicate involvement of IGFs in colorectal tumorigenesis. We investigated the CRC risk associated with 24 single nucleotide polymorphisms (SNPs) in 9 genes related to the IGF pathway and an IGF1 19-CA repeat polymorphism. Variants were selected from literature and genotyped in toenail DNA from 3,768 subcohort members and 2,580 CRC cases from the Netherlands Cohort Study, which has a case-cohort design (n = 120,852). We used the follow-up period 1986-2002. Eighteen SNPs were unequivocally associated with selected endpoints in the literature and unfavorable alleles were aggregated into a genetic sum score. Cox regression showed that a higher genetic sum score significantly increased CRC risk at all subsites, except the rectum, in men (highest vs. lowest tertile: HR for CRC = 1.36, 95% CI: 1.11, 1.65; P-trend = 0.002). Single SNPs (except the IGF1 SNP rs5742694) were not associated with risk. Models including the total number of IGF1 19-CA repeats showed CRC risk was halved at all subsites in women carrying < 38 repeats but not > 38 repeats (≤ 36 versus 38 repeats: HR for CRC = 0.44; 95% CI: 0.33, 0.58; P-trend < 0.001). These findings support a role for variants in IGF-related genes in colorectal tumorigenesis.
- Published
- 2015
- Full Text
- View/download PDF
25. Gene-diet interactions in exposure to heterocyclic aromatic amines and bulky DNA adduct levels in blood leukocytes.
- Author
-
Ho V, Peacock S, Massey TE, Godschalk RW, van Schooten FJ, Chen J, and King WD
- Subjects
- Adult, Aged, Amines toxicity, Diet, Environmental Exposure, Female, Humans, Male, Meat, Middle Aged, Sequence Analysis, DNA, Carcinogens toxicity, DNA Adducts, Gene-Environment Interaction, Heterocyclic Compounds toxicity, Leukocytes, Mononuclear physiology
- Abstract
Heterocyclic aromatic amines (HAAs), carcinogens produced in meat when cooked at high temperatures, are an emerging biologic explanation for the meat-colorectal cancer relationship. HAAs form DNA adducts; left unrepaired, adducts can induce mutations, which may initiate/promote carcinogenesis. The purpose of this research was to investigate the relationship between dietary HAAs, genetic susceptibility and bulky DNA adduct levels. Least squares regression was used to examine the relationship between dietary HAA exposure and bulky DNA adduct levels in blood measured using (32)P-postlabeling among 99 healthy volunteers. Gene-diet interactions between dietary HAAs and genetic factors relevant to the biotransformation of HAAs and DNA repair were also examined. No main effects of dietary HAAs on bulky DNA adduct levels was found. However, those with the putative NAT1 rapid acetylator phenotype had lower adduct levels than those with the slow acetylator phenotype (P = 0.02). Furthermore, having five or more 'at-risk' genotypes was associated with higher bulky DNA adduct levels (P = 0.03). Gene-diet interactions were observed between NAT1 polymorphisms and dietary HAAs (P < 0.05); among the slow acetylator phenotype, higher intakes of HAAs were associated with an increase in DNA adduct levels compared to lower intakes. This study provides evidence of a biologic relationship between dietary HAAs, genetic susceptibility and bulky DNA adduct formation. However, the lack of a strong main effect of HAAs suggests that dietary HAAs are not a large contributor to bulky DNA adducts in this population; future studies should consider relevant gene-diet interactions to clarify the role of HAAs in carcinogenesis., (© 2015 Wiley Periodicals, Inc.)
- Published
- 2015
- Full Text
- View/download PDF
26. Pulmonary Inflammation Impacts on CYP1A1-Mediated Respiratory Tract DNA Damage Induced by the Carcinogenic Air Pollutant Benzo[a]pyrene.
- Author
-
Arlt VM, Krais AM, Godschalk RW, Riffo-Vasquez Y, Mrizova I, Roufosse CA, Corbin C, Shi Q, Frei E, Stiborova M, van Schooten FJ, Phillips DH, and Spina D
- Subjects
- Animals, Male, Mice, Mice, Inbred C57BL, Respiratory System metabolism, Respiratory System pathology, Air Pollutants toxicity, Benzo(a)pyrene toxicity, Carcinogens toxicity, Cytochrome P-450 CYP1A1 metabolism, DNA Damage, Pneumonia enzymology
- Abstract
Pulmonary inflammation can contribute to the development of lung cancer in humans. We investigated whether pulmonary inflammation alters the genotoxicity of polycyclic aromatic hydrocarbons (PAHs) in the lungs of mice and what mechanisms are involved. To model nonallergic acute inflammation, mice were exposed intranasally to lipopolysaccharide (LPS; 20 µg/mouse) and then instilled intratracheally with benzo[a]pyrene (BaP; 0.5 mg/mouse). BaP-DNA adduct levels, measured by (32)P-postlabeling analysis, were approximately 3-fold higher in the lungs of LPS/BaP-treated mice than in mice treated with BaP alone. Pulmonary Cyp1a1 enzyme activity was decreased in LPS/BaP-treated mice relative to BaP-treated mice suggesting that pulmonary inflammation impacted on BaP-induced Cyp1a1 activity in the lung. Our results showed that Cyp1a1 appears to be important for BaP detoxification in vivo and that the decrease of pulmonary Cyp1a1 activity in LPS/BaP-treated mice results in a decrease of pulmonary BaP detoxification, thereby enhancing BaP genotoxicity (ie, DNA adduct formation) in the lung. Because less BaP was detoxified by Cyp1a1 in the lungs of LPS/BaP-treated mice, more BaP circulated via the blood to extrapulmonary tissues relative to mice treated with BaP only. Indeed, we observed higher BaP-DNA adduct levels in livers of LPS/BaP-treated mice compared with BaP-treated mice. Our results indicate that pulmonary inflammation could be a critical determinant in the induction of genotoxicity in the lung by PAHs like BaP. Cyp1a1 appears to be involved in both BaP bioactivation and detoxification although the contribution of other enzymes to BaP-DNA adduct formation in lung and liver under inflammatory conditions remains to be explored., (© The Author 2015. Published by Oxford University Press on behalf of the Society of Toxicology.)
- Published
- 2015
- Full Text
- View/download PDF
27. Environmental, dietary, maternal, and fetal predictors of bulky DNA adducts in cord blood: a European mother-child study (NewGeneris).
- Author
-
Pedersen M, Mendez MA, Schoket B, Godschalk RW, Espinosa A, Landström A, Villanueva CM, Merlo DF, Fthenou E, Gracia-Lavedan E, van Schooten FJ, Hoek G, Brunborg G, Meltzer HM, Alexander J, Nielsen JK, Sunyer J, Wright J, Kovács K, de Hoogh K, Gutzkow KB, Hardie LJ, Chatzi L, Knudsen LE, Anna L, Ketzel M, Haugen M, Botsivali M, Nieuwenhuijsen MJ, Cirach M, Toledano MB, Smith RB, Fleming S, Agramunt S, Kyrtopoulos SA, Lukács V, Kleinjans JC, Segerbäck D, and Kogevinas M
- Subjects
- Adult, Cohort Studies, Drinking Water chemistry, Europe, Female, Fetal Blood, Humans, Infant, Newborn, Male, Maternal-Fetal Exchange, Nitrogen Dioxide toxicity, Particulate Matter toxicity, Pregnancy, Trihalomethanes toxicity, Air Pollutants toxicity, DNA Adducts blood, Diet, Maternal Exposure adverse effects, Prenatal Exposure Delayed Effects blood
- Abstract
Background: Bulky DNA adducts reflect genotoxic exposures, have been associated with lower birth weight, and may predict cancer risk., Objective: We selected factors known or hypothesized to affect in utero adduct formation and repair and examined their associations with adduct levels in neonates., Methods: Pregnant women from Greece, Spain, England, Denmark, and Norway were recruited in 2006-2010. Cord blood bulky DNA adduct levels were measured by the 32P-postlabeling technique (n = 511). Diet and maternal characteristics were assessed via questionnaires. Modeled exposures to air pollutants and drinking-water disinfection by-products, mainly trihalomethanes (THMs), were available for a large proportion of the study population., Results: Greek and Spanish neonates had higher adduct levels than the northern European neonates [median, 12.1 (n = 179) vs. 6.8 (n = 332) adducts per 108 nucleotides, p < 0.001]. Residence in southern European countries, higher maternal body mass index, delivery by cesarean section, male infant sex, low maternal intake of fruits rich in vitamin C, high intake of dairy products, and low adherence to healthy diet score were statistically significantly associated with higher adduct levels in adjusted models. Exposure to fine particulate matter and nitrogen dioxide was associated with significantly higher adducts in the Danish subsample only. Overall, the pooled results for THMs in water show no evidence of association with adduct levels; however, there are country-specific differences in results with a suggestion of an association in England., Conclusion: These findings suggest that a combination of factors, including unknown country-specific factors, influence the bulky DNA adduct levels in neonates.
- Published
- 2015
- Full Text
- View/download PDF
28. Canonical autophagy does not contribute to cellular radioresistance.
- Author
-
Schaaf MB, Jutten B, Keulers TG, Savelkouls KG, Peeters HJ, van den Beucken T, van Schooten FJ, Godschalk RW, Vooijs M, and Rouschop KM
- Subjects
- Adenine analogs & derivatives, Adenine pharmacology, Cell Line, Tumor, Chloroquine pharmacology, DNA Repair drug effects, Humans, Phosphorylation, Radiation Tolerance genetics, Radiation, Ionizing, Radiation-Sensitizing Agents pharmacology, Signal Transduction drug effects, Autophagy drug effects
- Abstract
Background: (Pre)clinical studies indicate that autophagy inhibition increases response to anti-cancer therapies. Although promising, due to contradicting reports, it remains unclear if radiation therapy changes autophagy activity and if autophagy inhibition changes the cellular intrinsic radiosensitivity. Discrepancies may result from different assays and models through off-target effects and influencing other signaling routes. In this study, we directly compared the effects of genetic and pharmacological inhibition of autophagy after irradiation in human cancer cell lines., Materials and Methods: Changes in autophagy activity after ionizing radiation (IR) were assessed by flux analysis in eight cell lines. Clonogenic survival, DNA damage (COMET-assay) and H2AX phosphorylation were assessed after chloroquine or 3-methyladenine pretreatment and after ATG7 or LC3b knockdown., Results: IR failed to induce autophagy and chloroquine failed to change intrinsic radiosensitivity of cells. Interestingly, 3-methyladenine and ATG7- or LC3b-deficiency sensitized cancer cells to irradiation. Surprisingly, the radiosensitizing effect of 3-methyladenine was also observed in ATG7 and LC3b deficient cells and was associated with attenuated γ-H2AX formation and DNA damage repair., Conclusion: Our data demonstrate that the anti-tumor effects of chloroquine are independent of changes in intrinsic radioresistance. Furthermore, ATG7 and LC3b support radioresistance independent of canonical autophagy that involves lysosomal degradation., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
29. Polymorphisms in genes of the renin-angiotensin-aldosterone system and renal cell cancer risk: interplay with hypertension and intakes of sodium, potassium and fluid.
- Author
-
Deckers IA, van den Brandt PA, van Engeland M, van Schooten FJ, Godschalk RW, Keszei AP, and Schouten LJ
- Subjects
- Aged, Angiotensinogen genetics, Carcinoma, Renal Cell diet therapy, Carcinoma, Renal Cell pathology, DNA, Neoplasm genetics, Female, Follow-Up Studies, Gene-Environment Interaction, Humans, Hypertension diet therapy, Hypertension genetics, Kidney Neoplasms diet therapy, Kidney Neoplasms pathology, Male, Middle Aged, Peptidyl-Dipeptidase A genetics, Polymerase Chain Reaction, Prognosis, Prospective Studies, Receptor, Angiotensin, Type 1 genetics, Carcinoma, Renal Cell etiology, Hypertension complications, Kidney Neoplasms etiology, Polymorphism, Genetic genetics, Potassium, Dietary administration & dosage, Renin-Angiotensin System genetics, Sodium, Dietary administration & dosage
- Abstract
Hypertension is an established risk factor for renal cell cancer (RCC). The renin-angiotensin-aldosterone system (RAAS) regulates blood pressure and is closely linked to hypertension. RAAS additionally influences homeostasis of electrolytes (e.g. sodium and potassium) and fluid. We investigated single nucleotide polymorphisms (SNPs) in RAAS and their interactions with hypertension and intakes of sodium, potassium and fluid regarding RCC risk in the Netherlands Cohort Study (NLCS), which was initiated in 1986 and included 120,852 participants aged 55 to 69 years. Diet and lifestyle were assessed by questionnaires and toenail clippings were collected. Genotyping of toenail DNA was performed using the SEQUENOM® MassARRAY® platform for a literature-based selection of 13 candidate SNPs in seven key RAAS genes. After 20.3 years of follow-up, Cox regression analyses were conducted using a case-cohort approach including 3,583 subcohort members and 503 RCC cases. Two SNPs in AGTR1 were associated with RCC risk. AGTR1_rs1492078 (AA vs. GG) decreased RCC risk [hazard ratio (HR) (95% confidence interval (CI)): 0.70(0.49-1.00)], whereas AGTR1_rs5186 (CC vs. AA) increased RCC risk [HR(95%CI): 1.49(1.08-2.05)]. Associations were stronger in participants with hypertension. The RCC risk for AGT_rs3889728 (AG + AA vs. GG) was modified by hypertension (p interaction = 0.039). SNP-diet interactions were not significant, although HRs suggested interaction between SNPs in ACE and sodium intake. SNPs in AGTR1 and AGT influenced RCC susceptibility, and their effects were modified by hypertension. Sodium intake was differentially associated with RCC risk across genotypes of several SNPs, yet some analyses had probably inadequate power to show significant interaction. Results suggest that RAAS may be a candidate pathway in RCC etiology., (© 2014 UICC.)
- Published
- 2015
- Full Text
- View/download PDF
30. The shifting perception on antioxidants: the case of vitamin E and β-carotene.
- Author
-
Vrolijk MF, Opperhuizen A, Jansen EH, Godschalk RW, Van Schooten FJ, Bast A, and Haenen GR
- Subjects
- 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide pharmacology, Ascorbic Acid pharmacology, Cell Line, DNA Adducts biosynthesis, Epithelial Cells cytology, Epithelial Cells drug effects, Epithelial Cells metabolism, Glutathione S-Transferase pi metabolism, Humans, Oxidative Stress, Respiratory Mucosa cytology, Respiratory Mucosa drug effects, Respiratory Mucosa metabolism, Risk Factors, Antioxidants pharmacology, Glutathione S-Transferase pi antagonists & inhibitors, Vitamin E pharmacology, beta Carotene pharmacology
- Abstract
Antioxidants are vital for aerobic life, and for decades the expectations of antioxidants as health promoting agents were very high. However, relatively recent meta-analyses of clinical studies show that supplementation of antioxidants does not result in the presumed health benefit, but is associated with increased mortality. The dilemma that still needs to be solved is: what are antioxidants in the end, healthy or toxic? We have evaluated this dilemma by examining the presumed health effects of two individual antioxidants with opposite images i.e. the "poisonous" β-carotene and the "wholesome" vitamin E and focused on one aspect, namely their role in inducing BPDE-DNA adducts. It appears that both antioxidants promote DNA adduct formation indirectly by inhibition of the protective enzyme glutathione-S-transferase π (GST π). Despite their opposite image, both antioxidants display a similar type of toxicity. It is concluded that, in the appreciation of antioxidants, first their benefits should be identified and substantiated by elucidating their molecular mechanism. Subsequently, the risks should be identified including the molecular mechanism. The optimal benefit-risk ratio has to be determined for each antioxidant and each individual separately, also considering the dose., (Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
31. DNA from nails for genetic analyses in large-scale epidemiologic studies.
- Author
-
Hogervorst JG, Godschalk RW, van den Brandt PA, Weijenberg MP, Verhage BA, Jonkers L, Goessens J, Simons CC, Vermeesch JR, van Schooten FJ, and Schouten LJ
- Subjects
- Aged, Cohort Studies, Female, Genetic Testing, Genotype, Humans, Male, Middle Aged, Nails cytology, Prospective Studies, DNA genetics, Epidemiologic Studies, Nails metabolism
- Abstract
Background: Nails contain genomic DNA that can be used for genetic analyses, which is attractive for large epidemiologic studies that have collected or are planning to collect nail clippings. Study participants will more readily participate in a study when asked to provide nail samples than when asked to provide a blood sample. In addition, nails are easy and cheap to obtain and store compared with other tissues., Methods: We describe our findings on toenail DNA in terms of yield, quality, genotyping a limited set of SNPs with the Sequenom MassARRAY iPLEX platform and high-density genotyping with the Illumina HumanCytoSNP_FFPE-12 DNA array (>262,000 markers). We discuss our findings together with other studies on nail DNA and we compare nails and other frequently used tissue samples as DNA sources., Results: Although nail DNA is considerably degraded, genotyping a limited set of SNPs with the Sequenom MassARRAY iPLEX platform (average sample call rate, 97.1%) and high-density genotyping with the Illumina HumanCytoSNP_FFPE chip (average sample call rate, 93.8%) were successful., Conclusions: Nails are a suitable source of DNA for genotyping in large-scale epidemiologic studies, provided that methods are used that are suitable or optimized for degraded DNA. For genotyping through (next generation) sequencing where DNA degradation is less of an issue, nails may be an even more attractive DNA source, because it surpasses other sources in terms of ease and costs of obtaining and storing the samples., Impact: It is worthwhile to consider nails as a source of DNA for genotyping in large-scale epidemiologic studies. See all the articles in this CEBP Focus section, "Biomarkers, Biospecimens, and New Technologies in Molecular Epidemiology." Cancer Epidemiol Biomarkers Prev; 23(12); 2703-12. ©2014 AACR., (©2014 American Association for Cancer Research.)
- Published
- 2014
- Full Text
- View/download PDF
32. Hypoxia diminishes the detoxification of the environmental mutagen benzo[a]pyrene.
- Author
-
Schults MA, Sanen K, Godschalk RW, Theys J, van Schooten FJ, and Chiu RK
- Subjects
- 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide metabolism, Cell Hypoxia drug effects, Cell Line, Tumor, Chromatography, High Pressure Liquid, Culture Media chemistry, Cytochrome P-450 CYP1A1 metabolism, Cytochrome P-450 CYP1B1 metabolism, DNA Adducts metabolism, Dihydroxydihydrobenzopyrenes metabolism, Gene Expression Regulation, Enzymologic drug effects, Glucuronosyltransferase genetics, Glucuronosyltransferase metabolism, Humans, Inactivation, Metabolic drug effects, Kinetics, Oxygen pharmacology, Time Factors, Benzo(a)pyrene toxicity, Environmental Pollutants toxicity, Mutagens toxicity
- Abstract
Hypoxia promotes genetic instability and is therefore an important factor in carcinogenesis. We have previously shown that activation of the hypoxia responsive transcription factor HIFα can enhance the mutagenic phenotype induced by the environmental mutagen benzo[a]pyrene (BaP). To further elucidate the mechanism behind the ability of hypoxia to increase mutagenicity of carcinogens, we examined the activation and detoxification of BaP under hypoxic conditions. To this end, the human lung carcinoma cell line A549 was treated with BaP under 20%, 5% or 0.2% oxygen for 18h and alterations in BaP metabolism were assayed. First, BaP-induced expression of key metabolic enzymes was analysed; expression levels of the activating CYP1A1 and CYP1B1 were increased, while the detoxifying enzymes UGT1A6 and UGT2B7 were significantly reduced by hypoxia. To evaluate whether these changes had an effect on metabolism, levels of BaP and several of its metabolites were determined. Cells under hypoxia have a reduced capacity to metabolise BaP leaving more of the parent molecule intact. Additionally, BaP-7,8-dihydrodiol, the pre-cursor metabolite of the reactive metabolite BaP-7,8-dihydroxy-9,10-epoxide (BPDE), was formed in higher concentrations. Finally, under hypoxia, DNA adducts accumulated over a period of 168 h, whereas adducts were efficiently removed in 20% oxygen conditions. The delayed detoxification kinetics resulted in a 1.5-fold increase in DNA adducts. These data indicate that the metabolism under hypoxic conditions has shifted towards increased activation of BaP instead of detoxification and support the idea that modulation of carcinogen metabolism is an important additional mechanism for the observed HIF1 mediated genetic instability., (© The Author 2014. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2014
- Full Text
- View/download PDF
33. Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories.
- Author
-
Godschalk RW, Ersson C, Stępnik M, Ferlińska M, Palus J, Teixeira JP, Costa S, Jones GD, Higgins JA, Kain J, Möller L, Forchhammer L, Loft S, Lorenzo Y, Collins AR, van Schooten FJ, Laffon B, Valdiglesias V, Cooke M, Mistry V, Karbaschi M, Phillips DH, Sozeri O, Routledge MN, Nelson-Smith K, Riso P, Porrini M, López de Cerain A, Azqueta A, Matullo G, Allione A, and Møller P
- Subjects
- Adult, Calibration, Comet Assay, DNA Breaks, Double-Stranded, DNA-Formamidopyrimidine Glycosylase metabolism, Female, Humans, Middle Aged, Mutagenicity Tests, Regression Analysis, Cell Separation methods, DNA Damage, Laboratories, Leukocytes, Mononuclear metabolism
- Abstract
This study investigated the levels of DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG) sensitive sites, as assessed by the comet assay, in peripheral blood mononuclear cells (PBMC) from healthy women from five different countries in Europe. The laboratory in each country (referred to as 'centre') collected and cryopreserved PBMC samples from three donors, using a standardised cell isolation protocol. The samples were analysed in 13 different laboratories for DNA damage, which is measured by the comet assay. The study aim was to assess variation in DNA damage in PBMC samples that were collected in the same way and processed using the same blood isolation procedure. The inter-laboratory variation was the prominent contributor to the overall variation. The inter-laboratory coefficient of variation decreased for both DNA strand breaks (from 68 to 26%) and FPG sensitive sites (from 57 to 12%) by standardisation of the primary comet assay endpoint with calibration curve samples. The level of DNA strand breaks in the samples from two of the centres (0.56-0.61 lesions/10(6) bp) was significantly higher compared with the other three centres (0.41-0.45 lesions/10(6) bp). In contrast, there was no difference between the levels of FPG sensitive sites in PBMC samples from healthy donors in the different centres (0.41-0.52 lesion/10(6) bp)., (© The Author 2014. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2014
- Full Text
- View/download PDF
34. Redox and epigenetic regulation of the APE1 gene in the hippocampus of piglets: The effect of early life exposures.
- Author
-
Langie SA, Kowalczyk P, Tomaszewski B, Vasilaki A, Maas LM, Moonen EJ, Palagani A, Godschalk RW, Tudek B, van Schooten FJ, Berghe WV, Zabielski R, and Mathers JC
- Subjects
- 8-Hydroxy-2'-Deoxyguanosine, Animals, Animals, Newborn, CpG Islands, Deoxyguanosine analogs & derivatives, Deoxyguanosine metabolism, Epigenesis, Genetic, Female, Gene Expression Regulation, Developmental, Glutathione metabolism, Male, Oxidation-Reduction, Pregnancy, Promoter Regions, Genetic, Swine, Antioxidants pharmacology, DNA Methylation drug effects, DNA Repair, DNA-(Apurinic or Apyrimidinic Site) Lyase genetics, DNA-(Apurinic or Apyrimidinic Site) Lyase metabolism, Hippocampus metabolism
- Abstract
Oxidative stress via redox reactions can regulate DNA repair pathways. The base excision repair (BER) enzyme apurinic/apyrimidinic endonuclease 1 (APE1) is a key player in the redox regulation of DNA repair. Environmental factors can alter the methylation of DNA repair genes, change their expression and thus modulate BER activity and susceptibility to oxidative DNA damage. Therefore, we hypothesized that epigenetic modifications play a role in the redox regulation of APE1 in hippocampi of newborns and investigated the effect of supplementation of pregnant sows with a diet enriched in antioxidants and other nutrients on oxidative stress, DNA methylation and DNA repair in their offspring. High levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) and low levels of glutathione were detected in control piglets after birth compared with supplemented piglets, indicating the presence of oxidative stress. In control animals, this oxidative stress was associated with genomic DNA demethylation, decreased APE1 promoter methylation, increased APE1 expression and with slightly but not statistically significant increased BER-related DNA incision activity. Supplementation of piglets with antioxidants and other nutrients significantly lowered 8-oxodG levels compared to control animals, which was accompanied by overall lower APE1 promoter methylation and enhanced APE1 expression at day 7-28 after birth in supplemented piglets, although DNA incision activity was not significantly different between groups. Preliminary attempts to study the interaction between redox and epigenetic regulatory mechanisms revealed an inverse correlation between APE1 expression and methylation of CpG-sites 11 and 13 in the promoter region, which according to Genomatix "MatInspector" are located in the core binding sites of redox-sensitive transcription factors. We are the first to study methylation of the APE1 promoter and its role in mediating the functional effects of redox reactions induced by oxidative stress. Epigenetic and redox mechanisms may interact in regulating APE1-related DNA repair processes, involving redox-sensitive TFs., (Copyright © 2014 Elsevier B.V. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
35. Micronuclei in cord blood lymphocytes and associations with biomarkers of exposure to carcinogens and hormonally active factors, gene polymorphisms, and gene expression: the NewGeneris cohort.
- Author
-
Merlo DF, Agramunt S, Anna L, Besselink H, Botsivali M, Brady NJ, Ceppi M, Chatzi L, Chen B, Decordier I, Farmer PB, Fleming S, Fontana V, Försti A, Fthenou E, Gallo F, Georgiadis P, Gmuender H, Godschalk RW, Granum B, Hardie LJ, Hemminki K, Hochstenbach K, Knudsen LE, Kogevinas M, Kovács K, Kyrtopoulos SA, Løvik M, Nielsen JK, Nygaard UC, Pedersen M, Rydberg P, Schoket B, Segerbäck D, Singh R, Sunyer J, Törnqvist M, van Loveren H, van Schooten FJ, Vande Loock K, von Stedingk H, Wright J, Kleinjans JC, Kirsch-Volders M, and van Delft JH
- Subjects
- Carcinogens toxicity, Child, Cohort Studies, DNA Adducts adverse effects, DNA Adducts analysis, Europe epidemiology, Female, Fetal Blood chemistry, Gene Expression Profiling, Gene Expression Regulation drug effects, Genotype, Hormones adverse effects, Humans, Leukemia chemically induced, Malondialdehyde adverse effects, Malondialdehyde analysis, Micronucleus Tests, Pregnancy, Prenatal Exposure Delayed Effects chemically induced, T-Lymphocytes drug effects, Biomarkers analysis, Carcinogens analysis, Fetal Blood cytology, Hormones analysis, Leukemia epidemiology, Prenatal Exposure Delayed Effects epidemiology, T-Lymphocytes chemistry
- Abstract
Background: Leukemia incidence has increased in recent decades among European children, suggesting that early-life environmental exposures play an important role in disease development., Objectives: We investigated the hypothesis that childhood susceptibility may increase as a result of in utero exposure to carcinogens and hormonally acting factors. Using cord blood samples from the NewGeneris cohort, we examined associations between a range of biomarkers of carcinogen exposure and hormonally acting factors with micronuclei (MN) frequency as a proxy measure of cancer risk. Associations with gene expression and genotype were also explored., Methods: DNA and protein adducts, gene expression profiles, circulating hormonally acting factors, and GWAS (genome-wide association study) data were investigated in relation to genomic damage measured by MN frequency in lymphocytes from 623 newborns enrolled between 2006 and 2010 across Europe., Results: Malondialdehyde DNA adducts (M1dG) were associated with increased MN frequency in binucleated lymphocytes (MNBN), and exposure to androgenic, estrogenic, and dioxin-like compounds was associated with MN frequency in mononucleated lymphocytes (MNMONO), although no monotonic exposure-outcome relationship was observed. Lower frequencies of MNBN were associated with a 1-unit increase expression of PDCD11, LATS2, TRIM13, CD28, SMC1A, IL7R, and NIPBL genes. Gene expression was significantly higher in association with the highest versus lowest category of bulky and M1dG-DNA adducts for five and six genes, respectively. Gene expression levels were significantly lower for 11 genes in association with the highest versus lowest category of plasma AR CALUX® (chemically activated luciferase expression for androgens) (8 genes), ERα CALUX® (for estrogens) (2 genes), and DR CALUX® (for dioxins). Several SNPs (single-nucleotide polymorphisms) on chromosome 11 near FOLH1 significantly modified associations between androgen activity and MNBN frequency. Polymorphisms in EPHX1/2 and CYP2E1 were associated with MNBN., Conclusion: We measured in utero exposure to selected environmental carcinogens and circulating hormonally acting factors and detected associations with MN frequency in newborns circulating T lymphocytes. The results highlight mechanisms that may contribute to carcinogen-induced leukemia and require further research.
- Published
- 2014
- Full Text
- View/download PDF
36. You are what you eat, and so are your children: the impact of micronutrients on the epigenetic programming of offspring.
- Author
-
Vanhees K, Vonhögen IG, van Schooten FJ, and Godschalk RW
- Subjects
- DNA metabolism, DNA Methylation, Endocrine Disruptors toxicity, Female, Flavonoids pharmacology, Humans, Maternal-Fetal Relations, Oxidative Stress drug effects, Pregnancy, Epigenomics, Micronutrients metabolism
- Abstract
The research field of fetal programming has developed tremendously over the years and increasing knowledge suggests that both maternal and paternal unbalanced diet can have long-lasting effects on the health of offspring. Studies implicate that macronutrients play an important role in fetal programming, although the importance of micronutrients is also becoming increasingly apparent. Folic acid and vitamins B2, B6 and B12 are essential for one-carbon metabolism and are involved in DNA methylation. They can therefore influence the programming of the offspring's epigenome. Also, other micronutrients such as vitamins A and C, iron, chromium, zinc and flavonoids play a role in fetal programming. Since it is estimated that approximately 78 % of pregnant women in the US take vitamin supplements during pregnancy, more attention should be given to the long-term effects of these supplements on offspring. In this review we address several different studies which illustrate that an unbalanced diet prior and during pregnancy, regarding the intake of micronutrients of both mother and father, can have long-lasting effects on the health of adult offspring.
- Published
- 2014
- Full Text
- View/download PDF
37. Loss of VHL in RCC Reduces Repair and Alters Cellular Response to Benzo[a]pyrene.
- Author
-
Schults MA, Oligschlaeger Y, Godschalk RW, Van Schooten FJ, and Chiu RK
- Abstract
Mutations of the von Hippel-Lindau (VHL) tumor suppressor gene occur in the majority of sporadic renal-cell carcinomas (RCC). Loss of VHL function is associated with stabilization of hypoxia-inducible factor α (HIFα). We and others demonstrated that there is a two-way interaction between the aryl hydrocarbon receptor, which is an important mediator in the metabolic activation and detoxification of carcinogens, and the HIF1-pathway leading to an increased genetic instability when both pathways are simultaneously activated. The aim of this study was to investigate how environmental carcinogens, such as benzo[a]pyrene (BaP), which can be metabolically activated to BaP-7,8-diOH-9,10-epoxide (BPDE) play a role in the etiology of RCC. We exposed VHL-deficient RCC4 cells, in which HIFα is stabilized regardless of oxygen tension, to 0.1 μM BaP for 18 h. The mutagenic BPDE-DNA adduct levels were increased in HIFα stabilized cells. Using qRT-PCR, we demonstrated that absence of VHL significantly induced the mRNA levels of AhR downstream target CYP1A1. Furthermore, HPLC analysis indicated that loss of VHL increased the concentration of BaP-7,8-dihydroxydiol, the pre-cursor metabolite of BPDE. Interestingly, the capacity to repair BPDE-DNA adducts in the HIFα stabilized RCC4 cells, was markedly reduced. Taken together, these data indicate that loss of VHL affects BaP-mediated genotoxic responses in RCC and decreases repair capacity.
- Published
- 2013
- Full Text
- View/download PDF
38. Bulky dna adducts in cord blood, maternal fruit-and-vegetable consumption, and birth weight in a European mother-child study (NewGeneris).
- Author
-
Pedersen M, Schoket B, Godschalk RW, Wright J, von Stedingk H, Törnqvist M, Sunyer J, Nielsen JK, Merlo DF, Mendez MA, Meltzer HM, Lukács V, Landström A, Kyrtopoulos SA, Kovács K, Knudsen LE, Haugen M, Hardie LJ, Gützkow KB, Fleming S, Fthenou E, Farmer PB, Espinosa A, Chatzi L, Brunborg G, Brady NJ, Botsivali M, Arab K, Anna L, Alexander J, Agramunt S, Kleinjans JC, Segerbäck D, and Kogevinas M
- Subjects
- Female, Humans, Maternal Exposure adverse effects, Polycyclic Aromatic Hydrocarbons toxicity, Birth Weight physiology, DNA Adducts blood, Diet, Fetal Blood chemistry, Fruit, Vegetables
- Abstract
Background: Tobacco-smoke, airborne, and dietary exposures to polycyclic aromatic hydrocarbons (PAHs) have been associated with reduced prenatal growth. Evidence from biomarker-based studies of low-exposed populations is limited. Bulky DNA adducts in cord blood reflect the prenatal effective dose to several genotoxic agents including PAHs., Objectives: We estimated the association between bulky DNA adduct levels and birth weight in a multicenter study and examined modification of this association by maternal intake of fruits and vegetables during pregnancy., Methods: Pregnant women from Denmark, England, Greece, Norway, and Spain were recruited in 2006-2010. Adduct levels were measured by the 32P-postlabeling technique in white blood cells from 229 mothers and 612 newborns. Maternal diet was examined through questionnaires., Results: Adduct levels in maternal and cord blood samples were similar and positively correlated (median, 12.1 vs. 11.4 adducts in 108 nucleotides; Spearman rank correlation coefficient = 0.66, p < 0.001). Cord blood adduct levels were negatively associated with birth weight, with an estimated difference in mean birth weight of -129 g (95% CI: -233, -25 g) for infants in the highest versus lowest tertile of adducts. The negative association with birth weight was limited to births in Norway, Denmark, and England, the countries with the lowest adduct levels, and was more pronounced in births to mothers with low intake of fruits and vegetables (-248 g; 95% CI: -405, -92 g) compared with those with high intake (-58 g; 95% CI: -206, 90 g)., Conclusions: Maternal exposure to genotoxic agents that induce the formation of bulky DNA adducts may affect intrauterine growth. Maternal fruit and vegetable consumption may be protective.
- Published
- 2013
- Full Text
- View/download PDF
39. DNA-repair measurements by use of the modified comet assay: an inter-laboratory comparison within the European Comet Assay Validation Group (ECVAG).
- Author
-
Godschalk RW, Ersson C, Riso P, Porrini M, Langie SA, van Schooten FJ, Azqueta A, Collins AR, Jones GD, Kwok RW, Phillips DH, Sozeri O, Allione A, Matullo G, Möller L, Forchhammer L, Loft S, and Møller P
- Subjects
- Cell Line radiation effects, Environmental Monitoring, Humans, Monocytes cytology, Comet Assay methods, DNA Damage genetics, DNA Repair genetics, Monocytes radiation effects
- Abstract
The measurement of DNA-repair activity by extracts from cells or tissues by means of the single-cell gel electrophoresis (comet) assay has a high potential to become widely used in biomonitoring studies. We assessed the inter-laboratory variation in reported values of DNA-repair activity on substrate cells that had been incubated with Ro19-8022 plus light to generate oxidatively damaged DNA. Eight laboratories assessed the DNA-repair activity of three cell lines (i.e. one epithelial and two fibroblast cell lines), starting with cell pellets or with cell extracts provided by the coordinating laboratory. There was a large inter-laboratory variation, as evidenced by the range in the mean level of repair incisions between the laboratory with the lowest (0.002incisions/10(6)bp) and highest (0.988incisions/10(6)bp) incision activity. Nevertheless, six out of eight laboratories reported the same cell line as having the highest level of DNA-repair activity. The two laboratories that reported discordant results (with another cell line having the highest level of DNA-repair activity) were those that reported to have little experience with the modified comet assay to assess DNA repair. The laboratories were also less consistent in ordering the repair activity of the other two cell lines, probably because the DNA-repair activity by extracts from these cell lines were very similar (on average approximately 60-65% of the cell line with the highest repair capacity). A significant correlation was observed between the repair activity found in the provided and the self-made cell extracts (r=0.71, P<0.001), which indicates that the predominant source for inter-laboratory variation is derived from the incubation of the extract with substrate cells embedded in the gel. Overall, we conclude that the incubation step of cell extracts with the substrate cells can be identified as a major source of inter-laboratory variation in the modified comet assay for base-excision repair., (Copyright © 2013 Elsevier B.V. All rights reserved.)
- Published
- 2013
- Full Text
- View/download PDF
40. Maternal folate depletion and high-fat feeding from weaning affects DNA methylation and DNA repair in brain of adult offspring.
- Author
-
Langie SA, Achterfeldt S, Gorniak JP, Halley-Hogg KJ, Oxley D, van Schooten FJ, Godschalk RW, McKay JA, and Mathers JC
- Subjects
- 5-Methylcytosine metabolism, Animals, Base Sequence, Brain growth & development, Brain metabolism, DNA Damage, DNA Glycosylases genetics, DNA-Binding Proteins genetics, Diet, High-Fat, Dietary Fats administration & dosage, Female, Folic Acid administration & dosage, Gene Expression Regulation, Developmental drug effects, Male, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Pregnancy, Reverse Transcriptase Polymerase Chain Reaction, Vitamin B Complex administration & dosage, Vitamin B Complex pharmacology, Weaning, X-ray Repair Cross Complementing Protein 1, Brain drug effects, DNA Methylation drug effects, DNA Repair drug effects, Dietary Fats pharmacology, Folic Acid pharmacology, Maternal Nutritional Physiological Phenomena
- Abstract
The mechanisms through which environmental and dietary factors modulate DNA repair are still unclear but may include dysregulation of gene expression due to altered epigenetic markings. In a mouse model, we investigated the effect of maternal folate depletion during pregnancy and lactation, and high-fat feeding from weaning, on base excision repair (BER) and DNA methylation and expression of selected BER-related genes in the brain of adult offspring. While folate depletion did not affect BER activity of the mothers, BER increased in the offspring at weaning (P=0.052). In the long term, as observed in 6-mo-old offspring, the double insult, i.e., maternal low-folate supply and high-fat feeding from weaning, decreased BER activity significantly in the cortex, cerebellum, hippocampus, and subcortical regions (P≤0.017). This fall in BER activity was associated with small changes in methylation or expression of BER-related genes. Maternal folate depletion led to slightly increased oxidative DNA damage levels in subcortical regions of adult offspring, which may increase sensitivity to oxidative stress and predispose to neurological disorders. In summary, our data suggest that low-folate supply during early life may leave an epigenetic mark that can predispose the offspring to further dietary insults, causing adverse effects during adult life.
- Published
- 2013
- Full Text
- View/download PDF
41. Paternal lifestyle as a potential source of germline mutations transmitted to offspring.
- Author
-
Linschooten JO, Verhofstad N, Gutzkow K, Olsen AK, Yauk C, Oligschläger Y, Brunborg G, van Schooten FJ, and Godschalk RW
- Subjects
- Adult, Alleles, Child, DNA Mutational Analysis, Female, Gene Frequency, Humans, Income, Life Style, Male, Mutation Rate, Nuclear Family, Paternal Behavior, Pregnancy, Prospective Studies, Surveys and Questionnaires, Germ-Line Mutation, Intracellular Signaling Peptides and Proteins genetics, Minisatellite Repeats genetics, Smoking
- Abstract
Paternal exposure to high levels of radioactivity causes heritable germline minisatellite mutations. However, the effect of more general paternal exposures, such as cigarette smoking, on germline mutations remains unexplored. We analyzed two of the most commonly used minisatellite loci (CEB1 and B6.7) to identify germline mutations in blood samples of complete mother-father-child triads from the Norwegian Mother and Child Cohort Study (MoBa). The presence of mutations was subsequently related to general lifestyle factors, including paternal smoking before the partner became pregnant. Paternally derived mutations at the B6.7 locus (mutation frequency 0.07) were not affected by lifestyle. In contrast, high gross yearly income as a general measure of a healthy lifestyle coincided with low-mutation frequencies at the CEB1 locus (P=0.047). Income was inversely related to smoking behavior, and paternally derived CEB1 mutations were dose dependently increased when the father smoked in the 6 mo before pregnancy, 0.21 vs. 0.05 in smoking and nonsmoking fathers, respectively (P=0.061). These results suggest that paternal lifestyle can affect the chance of heritable mutations in unstable repetitive DNA sequences. To our knowledge, this is the first study reporting an effect of lifestyle on germline minisatellite mutation frequencies in a human population with moderate paternal exposures.
- Published
- 2013
- Full Text
- View/download PDF
42. DNA adducts and combinations of multiple lung cancer at-risk alleles in environmentally exposed and smoking subjects.
- Author
-
Peluso ME, Munnia A, Srivatanakul P, Jedpiyawongse A, Sangrajrang S, Ceppi M, Godschalk RW, van Schooten FJ, and Boffetta P
- Subjects
- DNA-(Apurinic or Apyrimidinic Site) Lyase genetics, Environmental Exposure, Epoxide Hydrolases genetics, Genetic Predisposition to Disease, Genetic Variation, Humans, NAD(P)H Dehydrogenase (Quinone) genetics, Risk, Superoxide Dismutase genetics, DNA Adducts, DNA Damage, Lung Neoplasms genetics, Smoking, Tobacco Smoke Pollution
- Abstract
Interindividual variation in DNA adduct levels in individuals exposed to similar amounts of environmental carcinogens may be due to genetic variability. We analysed the influence of genes involved in determining/modifying DNA damage, including microsomal epoxide hydrolase1 (EPHX1) His139Arg, N-acetyl-transferase, NAD(P)H:quinone oxidoreductase1 (NQO1) Pro187Ser, manganese superoxide dismutase2 (MnSOD2) Val16Ala, and apurinic/apyrimidinic endonuclease1 (APE1) Asp148Glu polymorphisms in blood of 120 smokers. Subsequently, we examined the effects of the combinations of the variant alleles of EPHX, NQO1 and MnSOD2 together with the wild type allele of APE1 on DNA damage by calculating the "sum of at-risk alleles." We reviewed the studies examining the relationships of DNA adducts with at-risk alleles in environmentally exposed subjects. Our findings showed that smokers carrying the EPHX1-139Arg and the NQO1-187Ser variants were significantly more likely to have higher adduct levels. Null associations were found with the other variants. Nevertheless, DNA adduct levels in smokers with ≥5 at-risk alleles were significantly different from those with fewer than two alleles. A similar picture emerged from studies of DNA adducts and at-risk alleles in environmentally exposed and smoking subjects. Certain at-risk allele combinations may confer a greater likelihood of increased levels of adducts after environmental insults. The increase in DNA adduct levels in susceptible subjects exposed to environmental carcinogens may reflect changes in the mechanisms that protect cells from the accumulation of genetic damage. Alterations of the physiological processes designed to maintain homeostasis may reduce the individual "genotoxic tolerance" to environmental challenges and result in phenotypes characterized by high levels of DNA adducts., (© 2013 Wiley Periodicals, Inc.)
- Published
- 2013
- Full Text
- View/download PDF
43. An ECVAG inter-laboratory validation study of the comet assay: inter-laboratory and intra-laboratory variations of DNA strand breaks and FPG-sensitive sites in human mononuclear cells.
- Author
-
Ersson C, Møller P, Forchhammer L, Loft S, Azqueta A, Godschalk RW, van Schooten FJ, Jones GD, Higgins JA, Cooke MS, Mistry V, Karbaschi M, Phillips DH, Sozeri O, Routledge MN, Nelson-Smith K, Riso P, Porrini M, Matullo G, Allione A, Stepnik M, Ferlińska M, Teixeira JP, Costa S, Corcuera LA, López de Cerain A, Laffon B, Valdiglesias V, Collins AR, and Möller L
- Subjects
- Adult, Dose-Response Relationship, Radiation, Female, Gamma Rays adverse effects, Humans, Leukocytes, Mononuclear radiation effects, Middle Aged, Reproducibility of Results, Comet Assay methods, DNA Breaks radiation effects, DNA-Formamidopyrimidine Glycosylase metabolism, Leukocytes, Mononuclear metabolism
- Abstract
The alkaline comet assay is an established, sensitive method extensively used in biomonitoring studies. This method can be modified to measure a range of different types of DNA damage. However, considerable differences in the protocols used by different research groups affect the inter-laboratory comparisons of results. The aim of this study was to assess the inter-laboratory, intra-laboratory, sample and residual (unexplained) variations in DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG)-sensitive sites measured by the comet assay by using a balanced Latin square design. Fourteen participating laboratories used their own comet assay protocols to measure the level of DNA strand breaks and FPG-sensitive sites in coded samples containing peripheral blood mononuclear cells (PBMC) and the level of DNA strand breaks in coded calibration curve samples (cells exposed to different doses of ionising radiation) on three different days of analysis. Eleven laboratories found dose-response relationships in the coded calibration curve samples on two or three days of analysis, whereas three laboratories had technical problems in their assay. In the coded calibration curve samples, the dose of ionising radiation, inter-laboratory variation, intra-laboratory variation and residual variation contributed to 60.9, 19.4, 0.1 and 19.5%, respectively, of the total variation. In the coded PBMC samples, the inter-laboratory variation explained the largest fraction of the overall variation of DNA strand breaks (79.2%) and the residual variation (19.9%) was much larger than the intra-laboratory (0.3%) and inter-subject (0.5%) variation. The same partitioning of the overall variation of FPG-sensitive sites in the PBMC samples indicated that the inter-laboratory variation was the strongest contributor (56.7%), whereas the residual (42.9%), intra-laboratory (0.2%) and inter-subject (0.3%) variations again contributed less to the overall variation. The results suggest that the variation in DNA damage, measured by comet assay, in PBMC from healthy subjects is assay variation rather than variation between subjects.
- Published
- 2013
- Full Text
- View/download PDF
44. Malondialdehyde-deoxyguanosine and bulky DNA adducts in schoolchildren resident in the proximity of the Sarroch industrial estate on Sardinia Island, Italy.
- Author
-
Peluso M, Munnia A, Ceppi M, Giese RW, Catelan D, Rusconi F, Godschalk RW, and Biggeri A
- Subjects
- Adolescent, Air Pollution, Child, Female, Humans, Islands, Italy, Male, Nasal Mucosa metabolism, Rural Population, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Urban Population, Volatile Organic Compounds adverse effects, DNA Adducts chemistry, Deoxyguanosine chemistry, Malondialdehyde chemistry, Occupational Exposure adverse effects
- Abstract
Air quality is a primary environmental concern in highly industrialised areas, with potential health effects in children residing nearby. The Sarroch industrial estate in Cagliari province, Sardinia Island, Italy, hosts the world's largest power plant and the second largest European oil refinery and petrochemical park. This industrial estate produces a complex mixture of air pollutants, including benzene, heavy metals and polycyclic aromatic hydrocarbons. Thus, we conducted a cross-sectional study to evaluate the prevalence of malondialdehyde-deoxyguanosine adducts in the nasal epithelium of 75 representative children, aged 6-14 years, attending primary and secondary schools in Sarroch in comparison with 73 rural controls. Additionally, the levels of bulky DNA adducts were analysed in a subset of 62 study children. DNA damage was measured by (32)P-postlabelling methodologies. The air concentrations of benzene and ethyl benzene were measured in the school gardens of Sarroch and a rural village by diffusive samplers. Outdoor measurements were also performed in other Sarroch areas and in the proximity of the industrial estate. The outdoor levels of benzene and ethyl benzene were significantly higher in the school gardens of Sarroch than in the rural village. Higher concentrations were also found in other Sarroch areas and in the vicinity of the industrial park. The mean levels of malondialdehyde-deoxyguanosine adducts per 10(8) normal nucleotides ± standard error (SE) were 74.6±9.1 and 34.1±4.4 in the children from Sarroch and the rural village, respectively. The mean ratio was 2.53, 95% confidence interval (CI): 1.71-2.89, P < 0.001, versus rural controls. Similarly, the levels of bulky DNA adducts per 10(8) normal nucleotides ± SE were 2.9±0.4 and 1.6±0.2 in the schoolchildren from Sarroch and the rural village, respectively. The means ratio was 1.90, 95% CI: 1.25-2.89, P = 0.003 versus rural controls. Our study indicates that children residing near the industrial estate have a significant increment of DNA damage.
- Published
- 2013
- Full Text
- View/download PDF
45. Intrauterine exposure to flavonoids modifies antioxidant status at adulthood and decreases oxidative stress-induced DNA damage.
- Author
-
Vanhees K, van Schooten FJ, van Waalwijk van Doorn-Khosrovani SB, van Helden S, Munnia A, Peluso M, Briedé JJ, Haenen GR, and Godschalk RW
- Subjects
- Animals, Anticarcinogenic Agents pharmacology, Antioxidants metabolism, Female, Flavonoids metabolism, Genistein pharmacology, Glutathione biosynthesis, Liver drug effects, Liver metabolism, Lung drug effects, Lung metabolism, Mice, Mice, Inbred C57BL, NF-E2-Related Factor 2 biosynthesis, Pregnancy, Quercetin pharmacology, Superoxide Dismutase biosynthesis, Antioxidants pharmacology, DNA Damage, Flavonoids pharmacology, Oxidative Stress, Prenatal Exposure Delayed Effects
- Abstract
Maternal intake of flavonoids, known for their antioxidant properties, may affect the offspring's susceptibility to developing chronic diseases at adult age, especially those related to oxidative stress, via developmental programming. Therefore, we supplemented female mice with the flavonoids genistein and quercetin during gestation, to study their effect on the antioxidant capacity of lung and liver of adult offspring. Maternal intake of quercetin increased the expression of Nrf2 and Sod2 in fetal liver at gestational day 14.5. At adult age, in utero exposure to both flavonoids resulted in the increased expression of several enzymatic antioxidant genes, which was more pronounced in the liver than in the adult lung. Moreover, prenatal genistein exposure induced the nonenzymatic antioxidant capacity in the adult lung, partly by increasing glutathione levels. Prenatal exposure to both flavonoids resulted in significantly lower levels of oxidative stress-induced DNA damage in liver only. Our observations lead to the hypothesis that a preemptive trigger of the antioxidant defense system in utero had a persistent effect on antioxidant capacity and as a result decreased oxidative stress-induced DNA damage in the liver., (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Published
- 2013
- Full Text
- View/download PDF
46. Benzo[a]pyrene-induced transcriptomic responses in primary hepatocytes and in vivo liver: toxicokinetics is essential for in vivo-in vitro comparisons.
- Author
-
van Kesteren PC, Zwart PE, Schaap MM, Pronk TE, van Herwijnen MH, Kleinjans JC, Bokkers BG, Godschalk RW, Zeilmaker MJ, van Steeg H, and Luijten M
- Subjects
- Animals, Apoptosis drug effects, Apoptosis genetics, Benzo(a)pyrene pharmacokinetics, Carcinogens pharmacokinetics, Cell Transformation, Neoplastic chemically induced, Cell Transformation, Neoplastic genetics, Cell Transformation, Neoplastic metabolism, Cell Transformation, Neoplastic pathology, Cells, Cultured, Computer Simulation, DNA Adducts metabolism, DNA Replication drug effects, Dose-Response Relationship, Drug, Hepatocytes metabolism, Hepatocytes pathology, High-Throughput Screening Assays, Liver metabolism, Liver pathology, Liver Neoplasms genetics, Liver Neoplasms metabolism, Liver Neoplasms pathology, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Models, Biological, Primary Cell Culture, Risk Assessment, Transcription, Genetic drug effects, Tumor Suppressor Protein p53 genetics, Xeroderma Pigmentosum Group A Protein genetics, Benzo(a)pyrene toxicity, Carcinogenicity Tests methods, Carcinogens toxicity, Gene Expression Profiling, Gene Expression Regulation, Neoplastic drug effects, Hepatocytes drug effects, Liver drug effects, Liver Neoplasms chemically induced
- Abstract
The traditional 2-year cancer bioassay needs replacement by more cost-effective and predictive tests. The use of toxicogenomics in an in vitro system may provide a more high-throughput method to investigate early alterations induced by carcinogens. Recently, the differential gene expression response in wild-type and cancer-prone Xpa (-/-) p53 (+/-) primary mouse hepatocytes after exposure to benzo[a]pyrene (B[a]P) revealed downregulation of cancer-related pathways in Xpa (-/-) p53 (+/-) hepatocytes only. Here, we investigated pathway regulation upon in vivo B[a]P exposure of wild-type and Xpa (-/-) p53 (+/-) mice. In vivo transcriptomics analysis revealed a limited gene expression response in mouse livers, but with a significant induction of DNA replication and apoptotic/anti-apoptotic cellular responses in Xpa (-/-) p53 (+/-) livers only. In order to be able to make a meaningful in vivo-in vitro comparison we estimated internal in vivo B[a]P concentrations using DNA adduct levels and physiologically based kinetic modeling. Based on these results, the in vitro concentration that corresponded best with the internal in vivo dose was chosen. Comparison of in vivo and in vitro data demonstrated similarities in transcriptomics response: xenobiotic metabolism, lipid metabolism and oxidative stress. However, we were unable to detect cancer-related pathways in either wild-type or Xpa (-/-) p53 (+/-) exposed livers, which were previously found to be induced by B[a]P in Xpa (-/-) p53 (+/-) primary hepatocytes. In conclusion, we showed parallels in gene expression responses between livers and primary hepatocytes upon exposure to equivalent concentrations of B[a]P. Furthermore, we recommend considering toxicokinetics when modeling a complex in vivo endpoint with in vitro models.
- Published
- 2013
- Full Text
- View/download PDF
47. Genetic polymorphisms in catalase and CYP1B1 determine DNA adduct formation by benzo(a)pyrene ex vivo.
- Author
-
Schults MA, Chiu RK, Nagle PW, Wilms LC, Kleinjans JC, van Schooten FJ, and Godschalk RW
- Subjects
- Adolescent, Adult, Aryl Hydrocarbon Hydroxylases metabolism, Carcinogens toxicity, Carcinogens, Environmental toxicity, Catalase metabolism, Cell Line, Tumor, Comet Assay, Cytochrome P-450 CYP1B1, DNA Repair drug effects, Epistasis, Genetic, Female, Gene Expression Regulation, Genetic Association Studies, Genotype, Humans, Linear Models, Lung cytology, Lung drug effects, Lung metabolism, Male, Middle Aged, NF-E2-Related Factor 2 genetics, NF-E2-Related Factor 2 metabolism, Oxidative Stress drug effects, Phenotype, Promoter Regions, Genetic, RNA, Messenger genetics, RNA, Messenger metabolism, Reverse Transcriptase Polymerase Chain Reaction, Young Adult, Aryl Hydrocarbon Hydroxylases genetics, Benzo(a)pyrene toxicity, Catalase genetics, DNA Adducts toxicity, Lymphocytes drug effects, Polymorphism, Single Nucleotide
- Abstract
Genetic polymorphisms can partially explain the large inter-individual variation in DNA adduct levels following exposure to polycyclic aromatic hydrocarbons. Effects of genetic polymorphisms on DNA adduct formation are difficult to assess in human studies because exposure misclassification attenuates underlying relationships. Conversely, ex vivo studies offer the advantage of controlled exposure settings, allowing the possibility to better elucidate genotype-phenotype relationships and gene-gene interactions. Therefore, we exposed lymphocytes of 168 non-smoking volunteers ex vivo to the environmental pollutant benzo(a)pyrene (BaP) and BaP-related DNA adducts were quantified. Thirty-four genetic polymorphisms were assessed in genes involved in carcinogen metabolism, oxidative stress and DNA repair. Polymorphisms in catalase (CAT, rs1001179) and cytochrome P450 1B1 (CYP1B1, rs1800440) were significantly associated with DNA adduct levels, especially when combined. Moreover, reverse transcription-polymerase chain reaction (RT-PCR) analysis in a subset of 30 subjects revealed that expression of catalase correlated strongly with expression of CYP1B1 (R = 0.92, P < 0.001). To further investigate the mechanism by which catalase influences CYP1B1 and how they simultaneously affect BaP-related DNA adduct levels, catalase expression was transiently knocked down in the human lung epithelial cell line A549. Although catalase knockdown did not immediately change CYP1B1 gene expression, recovery of catalase expression 8 h after the knockdown coincided with a 2.2-fold increased expression of CYP1B1 (P < 0.05). We conclude that the genetic polymorphism in the promoter region of CAT may determine the amount and activity of catalase, which may subsequently regulate the expression of CYP1B1. As a result, both genetic polymorphisms modulate DNA adduct levels in lymphocytes by BaP ex vivo.
- Published
- 2013
- Full Text
- View/download PDF
48. Organ specificity of beta-carotene induced lung gene-expression changes in Bcmo1-/- mice.
- Author
-
van Helden YG, Godschalk RW, van Schooten FJ, and Keijer J
- Subjects
- Adipose Tissue, White drug effects, Adipose Tissue, White metabolism, Animals, Down-Regulation, Extracellular Matrix Proteins genetics, Extracellular Matrix Proteins metabolism, Female, Frizzled Receptors genetics, Frizzled Receptors metabolism, Gene Expression Profiling methods, Genotype, Liver drug effects, Liver metabolism, Lung metabolism, Male, Mice, Mice, Knockout, Oligonucleotide Array Sequence Analysis methods, Organ Specificity drug effects, Sex Factors, Up-Regulation, Vitamin A Deficiency genetics, Vitamin A Deficiency physiopathology, Wnt Signaling Pathway, beta Carotene blood, beta-Carotene 15,15'-Monooxygenase genetics, beta-Carotene 15,15'-Monooxygenase metabolism, Dietary Supplements, Lung drug effects, beta Carotene administration & dosage
- Abstract
Scope: Whole genome transcriptome analysis of male and female beta-carotene 15,15'-monooxygenase knockout (Bcmo1(-/-) ) and Bcmo1(+/+) (wild-type) mice with or without 14 wk of BC supplementation was done. We previously showed that only 1.8% of the genes regulated by BC in lung were also regulated in liver and inguinal white adipose tissue (iWAT), suggesting lung specific responses. Here, we explicitly questioned the lung specificity., Methods and Results: We show that BC supplementation resulted in an opposite direction of gene-regulation in male compared to female Bcmo1(-/-) mice in lung, liver, and iWAT. This supports a systemic effect of BC on steroid hormone metabolism mediated responses. Lung, liver, and iWAT of female Bcmo1(-/-) mice showed an increased inflammatory response, which was counteracted by supplementation of BC. This supports a genotype dependent increased sensitivity of female mice for vitamin A deficiency. Finally, the effect of BC on Wnt signaling in male Bcmo1(-/-) mice was examined. Frizzled homolog 6 (Fzd6) downregulation was seen in all three tissues. Collagen triple helix containing 1 (Cthrc1) downregulation was seen in lung tissue only, suggesting specificity. Upregulation of genes involved in oxygen sensing was seen in lung and iWAT, while protocadherin upregulation was only seen in lung., Conclusion: Our results demonstrate that effects of BC are strongly sex dependent. While effects of BC on hormone metabolism mediated responses and inflammation are systemic, effects on Wnt signaling may be lung specific., (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2013
- Full Text
- View/download PDF
49. Aromatic DNA adducts and number of lung cancer risk alleles in Map-Ta-Phut Industrial Estate workers and nearby residents.
- Author
-
Peluso M, Srivatanakul P, Jedpiyawongse A, Sangrajrang S, Munnia A, Piro S, Ceppi M, Boffetta P, Godschalk RW, and van Schooten FJ
- Subjects
- Adult, Carcinogens toxicity, Case-Control Studies, Environmental Exposure adverse effects, Female, Genetic Predisposition to Disease, Humans, Lung Neoplasms chemically induced, Male, Polycyclic Aromatic Hydrocarbons toxicity, Polymorphism, Single Nucleotide, Thailand, DNA Adducts analysis, DNA-(Apurinic or Apyrimidinic Site) Lyase genetics, Epoxide Hydrolases genetics, Lung Neoplasms genetics, NAD(P)H Dehydrogenase (Quinone) genetics, Occupational Exposure adverse effects, Superoxide Dismutase genetics
- Abstract
The Map-Ta-Phut Industrial Estate (MIE) in Rayong, Thailand, is the location of one of the largest industrial complexes in southeastern Asia. The MIE complex produces a mixture of air pollutants, including polycyclic aromatic hydrocarbons, compounds capable to induce the generation of DNA adducts. DNA adducts are considered to be a biomarker of carcinogen exposure; however, its production can be modulated by genetic susceptibility. Thus, we analysed the influence of EPHX1 His139Arg (A>G, rs2234922) and NQO1 Pro187Ser (C>T, rs1800566) involved in the metabolism of polycyclic aromatic hydrocarbons; MnSOD(2) Val16Ala (C>T, rs1799725) a gene that acts against the free radical generation; APE1/Ref-1 Asp148Glu (T>G, rs3136820) a gene involved in the repair of DNA, and in the control of cell-cycle and apoptosis on leucocyte DNA adducts in 77 MIE workers, 69 Map-Ta-Phut residents, and 50 rural controls, Rayong, Thailand. We searched for associations with the 'sum of at-risk alleles' by combining the variant alleles of EPHX1, NQO1 and MnSOD(2) together with the wild-type allele of APE1, since they appeared to influence lung cancer risk. Although our findings revealed significant associations between DNA adducts and the EPHX1 His139Arg and NQO1 Pro187Ser polymorphisms, the combination of at-risk alleles was found to affect DNA damage much stronger. DNA adducts were significantly increased in the individuals bearing 4 and ≥ 5 at-risk alleles [mean ratio (MR) = 1.55, 95% CI 1.10-2.18, P = 0.012, and MR = 2.11, 95% CI 1.27-3.51, P = 0.004, respectively)]. After correction for residence/employment categorisation, a significant increment was present in the MIE workers with ≥ 5 alleles (MR = 2.88, 95% CI 1.46-5.71, P = 0.003). Our data indicate relationships between the generation of DNA adducts and the enzymatic activities of EPHX and NQO1. The combination of unfavourable genetic variants seems to determine the individuals' susceptibility, rather than a single polymorphism.
- Published
- 2013
- Full Text
- View/download PDF
50. Inter-laboratory variation in DNA damage using a standard comet assay protocol.
- Author
-
Forchhammer L, Ersson C, Loft S, Möller L, Godschalk RW, van Schooten FJ, Jones GD, Higgins JA, Cooke M, Mistry V, Karbaschi M, Collins AR, Azqueta A, Phillips DH, Sozeri O, Routledge MN, Nelson-Smith K, Riso P, Porrini M, Matullo G, Allione A, Stępnik M, Komorowska M, Teixeira JP, Costa S, Corcuera LA, López de Cerain A, Laffon B, Valdiglesias V, and Møller P
- Subjects
- Calibration, DNA-Formamidopyrimidine Glycosylase analysis, Endpoint Determination, Humans, Leukocytes, Mononuclear chemistry, Leukocytes, Mononuclear cytology, Linear Models, Comet Assay methods, Comet Assay standards, DNA Damage, Laboratories standards
- Abstract
There are substantial inter-laboratory variations in the levels of DNA damage measured by the comet assay. The aim of this study was to investigate whether adherence to a standard comet assay protocol would reduce inter-laboratory variation in reported values of DNA damage. Fourteen laboratories determined the baseline level of DNA strand breaks (SBs)/alkaline labile sites and formamidopyrimidine DNA glycosylase (FPG)-sensitive sites in coded samples of mononuclear blood cells (MNBCs) from healthy volunteers. There were technical problems in seven laboratories in adopting the standard protocol, which were not related to the level of experience. Therefore, the inter-laboratory variation in DNA damage was only analysed using the results from laboratories that had obtained complete data with the standard comet assay protocol. This analysis showed that the differences between reported levels of DNA SBs/alkaline labile sites in MNBCs were not reduced by applying the standard assay protocol as compared with the laboratory's own protocol. There was large inter-laboratory variation in FPG-sensitive sites by the laboratory-specific protocol and the variation was reduced when the samples were analysed by the standard protocol. The SBs and FPG-sensitive sites were measured in the same experiment, indicating that the large spread in the latter lesions was the main reason for the reduced inter-laboratory variation. However, it remains worrying that half of the participating laboratories obtained poor results using the standard procedure. This study indicates that future comet assay validation trials should take steps to evaluate the implementation of standard procedures in participating laboratories.
- Published
- 2012
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.