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RU Radboud Universiteit Nijmegen, 2 oktober 2007, Promotor : Krieken, J.H.J.M. van Co-promotores : Peters, W.H.M., Nagengast, F.M., Nagtegaal, I.D., Contains fulltext : 52518.pdf (publisher's version ) (Open Access)

Contains fulltext : 52107.pdf (publisher's version ) (Closed access), Current treatments of subfertile couples are usually empiric, as the true cause of subfertility often remains unknown. Therefore, we outline the role of nutritional and biochemical factors in reproduction and subfertility. A literature search was performed using MEDLINE, Science Direct and bibliographies of published work with both positive and negative results. The studies showed that folate has a role in spermatogenesis. In female reproduction, folate is also important for oocyte quality and maturation, implantation, placentation, fetal growth and organ development. Zinc has also been implicated in testicular development, sperm maturation and testosterone synthesis. In females, zinc plays a role in sexual development, ovulation and the menstrual cycle. Both folate and zinc have antioxidant properties that counteract reactive oxygen species (ROS). Thiols, such as glutathione, balance the levels of ROS produced by spermatozoa and influence DNA compaction and the stability and motility of spermatozoa. Oocyte maturation, ovulation, luteolysis and follicle atresia are also affected by ROS. After fertilization, glutathione is important for sperm nucleus decondensation and pronucleus formation. Folate, zinc, ROS and thiols affect apoptosis, which is important for sperm release, regulation of follicle atresia, degeneration of the corpus luteum and endometrial shedding. Therefore, the concentrations of these nutrients may have substantial effects on reproduction. In conclusion, nutritional and biochemical factors affect biological processes in male and female reproduction. Further research should identify pathways that may lead to improvements in care and treatment of subfertility.

Contains fulltext : 52234.pdf (publisher's version ) (Closed access), Primary carcinomas of the small intestine are rare and the mechanism of their pathogenesis is poorly understood. Patients with familial adenomatous polyposis (FAP) have a high risk of developing duodenal carcinomas. The aim of this study is to gain more insight into the development of duodenal carcinomas. Therefore, five FAP-related duodenal carcinomas were characterized for chromosomal and methylation alterations, which were compared to those observed in sporadic duodenal carcinomas. Comparative genomic hybridization (CGH) and methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) was performed in 10 primary sporadic and five primary FAP-related duodenal carcinomas. In the FAP-related carcinomas, frequent gains were observed on chromosomes 8, 17 and 19, whereas in sporadic carcinomas they occurred on chromosomes 8, 12, 13 and 20. In 60% of the sporadic carcinomas, gains in the regions of chromosome 12 were observed which were absent in the FAP-related carcinomas (P=0.04). Hypermethylation was observed in the immunoglobulin superfamily genes member 4 (IGSF4), TIMP metallopeptidase inhibitor 3 (TIMP3), Estrogen receptor 1 (ESR1), adenomatous polyposis coli (APC), H-cadherin (CDH13) and paired box gene 6 (PAX6) genes. Hypermethylation of PAX6 was only observed in FAP-related carcinomas (3/5) and not in sporadic carcinomas (P=0.02). In conclusion, in contrast to sporadic duodenal carcinomas, gains on chromosome 12 were not observed in duodenal carcinomas of patients with FAP. Identification of the genes in these regions of chromosome 12 could lead to a better understanding of the carcinogenesis pathways leading to sporadic and FAP-related duodenal carcinomas. Furthermore, hypermethylation seems to be a general feature of both FAP-related duodenal carcinomas as well as sporadic duodenal carcinomas with the exception of the PAX6 gene, which is methylated only in FAP-related carcinomas.

Contains fulltext : 53015.pdf (publisher's version ) (Closed access), BACKGROUND/AIMS: Probably no single gene is responsible for pre-eclampsia, but the disease merely is the result of polymorphisms in several genes in association with environmental factors. We therefore studied the simultaneous occurrence of several genetic polymorphisms in biotransformation enzymes in women who had developed pre- eclampsia, either with or without the HELLP syndrome, in comparison with healthy controls. METHODS: The results of two previous studies on genetic polymorphisms in glutathione S-transferases P1, M1 and T1, epoxide hydrolase (EPHX) and cytochrome P4501A1 (CYP1A1) in 167 women with a history of pre-eclampsia and in 110 controls were combined. chi(2) analyses were used for statistical evaluation of the number of polymorphisms between cases and controls. RESULTS: There was a significant association with the number of genetic polymorphisms in biotransformation enzymes, pointing at an increased toxification or decreased detoxification, in women with a history of pre-eclampsia, as compared to healthy controls (p < 0.001). CONCLUSION: Women withthe simultaneous occurrence of two or more genetic polymorphisms in the above-mentioned biotransformation enzymes, most probably resulting in a disturbed detoxification capacity, may be at increased risk for pre-eclampsia.

Contains fulltext : 53105.pdf (publisher's version ) (Open Access), To eliminate the risk of colorectal cancer in patients with familial adenomatous polyposis (FAP), reconstructive proctocolectomy is performed. Although most colonic mucosa is resected during the ileal pouch anal anastomosis, adenomas and carcinomas may develop in the pouch. This may be caused by altered cell kinetics due to intraluminal changes in the pouch. In 32 patients with FAP, biopsy specimens from the mucosa of the pouch and also of the afferent ileal loop were taken. Tissue sections were immunohistochemically processed with the monoclonal antibodies M30 and MIB-1 to assess apoptotic and proliferative indices, respectively. Cell proliferation was also assessed by a modified sign test. There were no significant differences in apoptotic rates between the mucosa of the pouch and the mucosa of the afferent ileal loop. However, cell proliferation was significantly higher in the mucosa of the pouch vs afferent ileal loop, both by using the quantitative (68.3% vs 61.6%, p = 0.001) and semiquantitative methods (p < 0.05). Our newly developed semiquantitative approach outperformed previously described methods. The higher cell proliferation in the pouch as compared to the afferent ileal loop may contribute to the increased risk for adenomas and carcinomas in the pouch of patients with FAP and emphasizes the need for regular endoscopic surveillance.

Contains fulltext : 51721.pdf (publisher's version ) (Closed access), Epidemiological studies have shown that ingestion of isoflavone-rich soy products is associated with a reduced risk for the development of breast cancer. In the present study, we investigated the hypothesis that genistein modulates the expression of glutathione S-transferases (GSTs) in human breast cells, thus conferring protection towards genotoxic carcinogens which are GST substrates. Our approach was to use human mammary cell lines MCF-10A and MCF-7 as models for non-neoplastic and neoplastic epithelial breast cells, respectively. MCF-10A cells expressed hGSTA1/2, hGSTA4-4, hGSTM1-1 and hGSTP1-1 proteins, but not hGSTM2-2. In contrast, MCF-7 cells only marginally expressed hGSTA1/2, hGSTA4-4 and hGSTM1-1. Concordant to the protein expression, the hGSTA4 and hGSTP1 mRNA expression was higher in the non-neoplastic cell line. Exposure to genistein significantly increased hGSTP1 mRNA (2.3-fold), hGSTP1-1 protein levels (3.1-fold), GST catalytic activity (4.7-fold) and intracellular glutathione concentrations (1.4-fold) in MCF-10A cells, whereas no effects were observed on GST expression or glutathione concentrations in MCF-7 cells. Preincubation of MCF-10A cells with genistein decreased the extent of DNA damage by 4-hydroxy-2-nonenal (150 microM) and benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide (50 microM), compounds readily detoxified by hGSTA4-4 and hGSTP1-1. In conclusion, genistein pretreatment protects non-neoplastic mammary cells from certain carcinogens that are detoxified by GSTs, suggesting that dietary-mediated induction of GSTs may be a mechanism contributing to prevention against genotoxic injury in the aetiology of breast cancer.

Contains fulltext : 51504.pdf ( ) (Open Access), BACKGROUND: Glutathione is a tripeptide composed of glutamate, cysteine and glycine, accomplishing a broad range of vital functions. Synthesis of glutathione and cysteine is performed mainly in the liver, whereas most other tissues are supplied with these thiols via sinusoidal efflux into the blood. Since canalicular efflux also occurs, thiols may be present in human bile. However, thiol composition of human gallbladder bile is largely unknown, which makes it difficult to speculate on the exact function of thiols in bile. In this study we report on the levels of non-protein bound thiols in gallbladder bile of patients with various gastrointestinal disorders. METHODS: Gallbladder bile was obtained after cholecystectomy from 30 patients who were operated for pancreatic cancer, duodenal cancer, chronic pancreatitis or cholecystolithiasis. Bile was analysed for non-protein bound total- and oxidised glutathione and related thiols, by high performance liquid chromatography. RESULTS: A more than 100-fold inter-individual variation in non-protein bound thiol levels was found in human gallbladder bile of patients with a variety of gastrointestinal disorders. Bile did contain high amounts of cysteine, whereas much lower levels of glutathione, cysteinylglycine and homocysteine were detected. Most thiols were present in their oxidised forms. CONCLUSION: Thiols are present in considerable amounts in human gallbladder bile of patients with various gastrointestinal disorders, levels of cysteine being much higher than those of glutathione and other thiols. Most thiols were in their oxidised forms, which may indicate the presence of considerable chemical- or oxidative stress in the patients studied here.

Contains fulltext : 52432schaik.pdf (publisher's version ) (Closed access)

Contains fulltext : 51701.pdf (publisher's version ) (Closed access), High glutathione S-transferase (GST) activity may contribute to colorectal cancer prevention. Functional polymorphisms are known in the GSTM1, GSTT1, GSTA1 and GSTP1 genes. The influence of these GST polymorphisms and recent fruit and vegetable consumption on GST levels and activity has not been investigated simultaneously in a human population. Also, it is not clear if blood GST activity reflects rectal GST activity. Therefore, we determined GST polymorphisms in 94 patients scheduled for sigmoidoscopy. Rectal GST isoenzyme levels (GSTM1, GSTM2, GSTT1, GSTA and GSTP1) were measured by quantitative western blotting, and rectal and white blood cell total GST activities were measured spectrophotometrically using 1-chloro-2,4-dinitrobenzene (CDNB) as a substrate. Vegetable and fruit consumption was assessed by dietary record. As expected, the GSTM1 and GSTT1 deletion polymorphisms, and the GSTA1 g.-69C-->T polymorphism significantly affected the respective isoenzyme levels. Also, rectal GST isoenzyme levels differed between those with and without recent consumption of Alliaceae, Cucurbitaceae, Apiaceae and citrus fruit. Rectal GST activity, however, was not clearly influenced by fruit and vegetable consumption. It was most significantly determined by the GSTP1 c.313A-->G polymorphism; compared with the 313AA genotypes, the 313AG and 313GG genotypes showed 36 and 67 nmol/min/mg protein (P < 0.001) lower GST activity, respectively. The correlation between rectal and white blood cell GST activities was low (r = 0.40, P < 0.001), and the relevance of the various genetic and dietary factors appeared to differ between the two tissues. In conclusion, this study indicates that the GST enzyme system is influenced by both GST polymorphisms and consumption of fruits and vegetables. The latter appeared more important for individual rectal GST isoenzyme levels than for total GST activity, which could affect detoxification of isoenzyme-specific substrates. The study results do no sup

Contains fulltext : 51690.pdf (publisher's version ) (Closed access), BACKGROUND: Liver failure following liver surgery is caused by an insufficient functioning remnant cell mass. This can be due to insufficient liver volume and can be aggravated by additional cell death during or after surgery. The aim of this study was to elucidate the causes of hepatocellular injury in patients undergoing liver resection. METHODS: Markers of hepatocyte injury (AST, GSTalpha, and L-FABP) and inflammation (IL-6) were measured in plasma of patients undergoing liver resection with and without intermittent inflow occlusion. To study the separate involvement of the intestines and the liver in systemic L-FABP release, arteriovenous concentration differences for L-FABP were measured. RESULTS: During liver manipulation, liver injury markers increased significantly. Arterial plasma levels and transhepatic and transintestinal concentration gradients of L-FABP indicated that this increase was exclusively due to hepatic and not due to intestinal release. Intermittent hepatic inflow occlusion, anesthesia, and liver transection did not further enhance arterial L-FABP and GSTalpha levels. Hepatocyte injury was followed by an inflammatory response. CONCLUSIONS: This study shows that liver manipulation is a leading cause of hepatocyte injury during liver surgery. A potential causal relation between liver manipulation and systemic inflammation remains to be established; but since the inflammatory response is apparently initiated early during major abdominal surgery, interventions aimed at reducing postoperative inflammation and related complications should be started early during surgery or beforehand.

Contains fulltext : 50160.pdf (publisher's version ) (Closed access), Colorectal cancer (CRC) is one of the most common forms of cancer in Western countries. CRC has been associated with genetic and lifestyle factors. Individual susceptibility to CRC may be due partly to variations in detoxification capacity in the gastrointestinal tract. Genetic polymorphisms in detoxification enzymes may result in variations in detoxification activities, which subsequently might influence the levels of toxic/carcinogenic compounds, and this may influence the risk for CRC. Therefore, we determined whether polymorphisms in the genes coding for microsomal epoxide hydrolase (EPHX1), NAD(P)H:quinone oxidoreductase (NQO1), cytochrome P450 2E1 (CYP2E1) and alcohol dehydrogenase (ADH3) predispose to the development of CRC. DNA samples were obtained from 371 patients with sporadic CRC and 415 healthy controls. Patients and controls were all of Caucasian origin. All genetic polymorphisms were determined by polymerase chain reaction, eventually followed by restriction-fragment-length-polymorphism analyses, except for the EPHX1 codon 113 polymorphism, which was genotyped by an allele-specific discrimination assay. Calculation of crude Odds Ratios (ORs) revealed an increased risk for CRC associated with variant NQO1 (OR 1.5, 95% CI 1.1-2.0) and CYP2E1 intron 6 genotypes (OR 2.2, 95% CI 1.3-3.8). However, after adjustment for age and gender, logistic regression analyses only showed a statistically significant risk for CRC associated with variant NQO1 genotypes (OR 1.6, 95% CI 1.03-2.4). No associations were found between CRC and the other polymorphic genes as mentioned above. In conclusion, these data suggest that the presence of variant NQO1 genotypes, with expected reduced enzyme activities might enhance susceptibility to CRC.

Contains fulltext : 50774.pdf (publisher's version ) (Open Access) Contains fulltext : 50774_pub.pdf (publisher's version ) (Open Access), AIM: To assess the levels of antioxidant capacity and oxidative damage in blood of chronic pancreatitis (CP) patients in comparison with those in healthy control subjects, by using several different analytical techniques. METHODS: Thirty-five CP patients and 35 healthy control subjects were investigated prospectively with respect to plasma levels of thiols, ferric reducing ability of plasma (FRAP, i.e. antioxidant capacity), levels of protein carbonyls and thiobarbituric acid reactive substances (TBARS). Additionally, we evaluated the production of reactive oxygen species (ROS) in whole blood. RESULTS: The antioxidative thiols including cysteine, cysteinylglycine and glutathione were significantly lower in CP patients. In addition, the non-enzymatic antioxidant capacity was significantly lower in CP patients, which correlated with the amount of oxidative protein (protein carbonyls) and the extent of lipid damage (TBARS), both were significantly higher in CP patients. The ROS production in whole blood after stimulation with phorbol 12-myritate 13-acetaat, demonstrated a strong tendency to produce more ROS in CP patients. CONCLUSION: Oxidative stress may contribute to the pathogenesis of chronic pancreatitis by decreasing antioxidant capacity and increasing oxidative damage in CP patients may be a rationale for intervention with antioxidant therapy.

Contains fulltext : 50088.pdf (publisher's version ) (Closed access), CONCLUSIONS: Inter-regional differences in the distribution of genetic polymorphisms in glutathione S-transferases (GSTs) exist, which may have significant effect on the outcome of other GST polymorphism studies. The GSTT1 null genotype appears to be involved in modulation of the risk for head and neck squamous cell carcinoma (HNSCC). BACKGROUND: The risk of HNSCC is strongly associated with smoking of cigarettes and consumption of alcohol, resulting in a load of toxins/carcinogens. Detoxification of such exogenous harmful compounds often occurs by phase II enzymes such as GSTs. Proper functioning of these enzymes may be deficient due to the presence of particular genetic polymorphisms in these GSTs, and this may increase the risk for HNSCC. We compared the GSTT1, GSTM1 and GSTP1 genotype frequencies in two groups of healthy blood donors, collected from different but adjacent regions in the Netherlands, with those of a group of patients with HNSCC. SUBJECTS AND METHODS: The GSTM1,GSTT1 and GSTP1 genotype frequencies in two Dutch Caucasian control populations (n = 207 and n = 285) from different but adjacent geographical regions (Maastricht and Nijmegen; distance, 125 km) and 185 patients with HNSCC from the Maastricht region were determined by PCR-related methods. RESULTS: For the occurrence of the GSTT1 null genotype we found a significant difference (p=0.003) between the two control groups (20.3% vs 33.0% null genotype in the Nijmegen and Maastricht control groups, respectively). Since the HNSCC patients were collected from the Maastricht area, comparison with the Maastricht controls reveals a significant difference for GSTT1 null rates, which are lower in patients vs controls (OR = 0.49, CI = 0.32-0.76).

Contains fulltext : 49315.pdf (publisher's version ) (Open Access), Glutathione S-transferases (GST) are a family of enzymes involved in phase-II detoxification of endogenous and xenobiotic compounds. Polymorphisms in GST genes have been associated with susceptibility to different diseases. In this study we determined the frequencies of polymorphisms in GSTM1, GSTT1, and GSTP1 in DNA of 138 children from Cameroon, presenting with uncomplicated malaria (N = 19), malaria with minor complications (N = 81), or severe malaria (N = 38). Analyses of GSTM1 and GSTT1 were performed using PCR-multiplex procedure, while GSTP1 was done by PCR-RFLP. Subjects presenting with malaria with complications were found more often of the GSTM1-null genotype (58-64%) as compared with those with uncomplicated malaria (32%), a difference that was statistically significant. We conclude that the GSTM1-null genotype is associated with malaria with complications.

Contains fulltext : 51210.pdf (publisher's version ) (Closed access), BACKGROUND: To study the possible involvement of an (im)balance between oxidants and antioxidants in pre-eclampsia concentrations of intra- and extracellular blood antioxidants in women with uncomplicated and hypertensive pregnancies, they were studied preconceptionally and throughout pregnancy. METHODS: In uncomplicated pregnancies (n = 19) and hypertensive pregnancies (n = 6) concentrations of whole blood and plasma thiols, plasma vitamins/E and C, hemoglobin, and hematocrit were assessed at preconception, 6, 10, 20, and 37 weeks of gestational age, as well as six weeks postpartum. A repeated mixed model was used for statistical analysis. RESULTS: Vitamin C and most whole blood and plasma thiol concentrations decreased during pregnancy, while vitamin E, whole blood oxidized cysteinyl-glycine and the ratio of free to oxidized homocysteine revealed a linear increase during pregnancy. Postpartum plasma cysteine and vitamin C levels and the ratio of free to oxidized levels of cysteine, cysteinyl-glycine, and glutathione were significantly (p <0.05) lower as compared to preconceptional levels, whereas whole blood oxidized cysteine, cysteinyl-glycine and glutathione levels, and whole blood and plasma homocysteine levels were significantly (p <0.05) higher six weeks after delivery. Plasma cysteine and homocysteine, and whole blood oxidized cysteine and homocysteine levels were significantly (p <0.05) higher at 37 weeks of gestational age in the hypertensive group compared to those in the uncomplicated group. There were no other differences between the hypertensive and uncomplicated groups. CONCLUSION: In normal pregnancy there seems a balance between antioxidant and oxidant concentrations despite modest oxidative stress. In mildly hypertensive pregnancies a marginal imbalance may occur.

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Contains fulltext : 50383.pdf (publisher's version ) (Closed access), Early placental development is characterised by rapid cell differentiation and migration, matrix remodelling and angiogenesis. The enzyme NAD(P)H oxidase is a major source of superoxide anions implicated in signalling pathways regulating these processes in other systems. It is also thought to be involved in oxygen sensing and regulation of the expression of antioxidant genes. We therefore investigated NAD(P)H oxidase activity in placental tissues in early pregnancy and at term, and correlated this with antioxidant capacity. We collected placental tissues from women undergoing termination of pregnancy (n=19; gestational age 11(+6)+/-1(+0) weeks), and those with elective caesarean section at term after uncomplicated pregnancy (n=15; gestational age 38(+6)+/-0(+4) weeks). Tissues were assayed for superoxide production, using lucigenin chemiluminescence, and three independent markers of antioxidant capacity. In human placentas from normal deliveries at term substantial basal NAD(P)H activity was present. Activity was almost threefold higher in early pregnancy (P<0.0001). This was paralleled by higher total antioxidant capacity (P<0.0001), tissue glutathione concentrations (P<0.01) and gluthathione S-transferase enzyme activity (P<0.05) when compared to corresponding term placental values. NAD(P)H oxidase mediated superoxide generation could be an important modulator of the antioxidant defence response in early pregnancy.

Contains fulltext : 49593.pdf (publisher's version ) (Closed access), BACKGROUND & AIMS: Patients on total parenteral nutrition depend on the composition of the nutritional formulation for maintenance of their oxidant-antioxidant balance. The present observational study was conducted to evaluate a substantial part of our patient population for evidence of oxidative stress. METHODS: Venous blood samples were obtained from 41 patients on home parenteral nutrition (HPN) and 41 healthy controls. Glutathione in plasma and whole blood, glutathione peroxidase and superoxide dismutase in erythrocytes and total plasma antioxidant capacity were measured to assess the antioxidant status. Oxidant status was evaluated by measuring the production of reactive oxygen species by leukocytes. Oxidative damage was assessed by measuring lipid peroxidation and protein oxidation products. RESULTS: Patients on HPN showed some signs of increased oxidative stress, however, there were no signs for oxidative damage, compared with healthy controls. In addition, activity of any underlying disease was not associated with increased oxidative stress. CONCLUSIONS: The current treatment regime for patients on HPN at our center apparently prevents the development of significant oxidative damage, despite signs of some oxidative stress. Based on these data, adaptations in the composition of parenteral nutritional formulations do not seem mandatory.

Contains fulltext : 49807.pdf (publisher's version ) (Closed access), BACKGROUND: Small intestinal malignancies in humans are rare; however, patients with coeliac disease have a relatively high risk for such tumours. Intestinal UDP-glucuronosyltransferases are phase II drug metabolism enzymes also involved in the detoxification of ingested toxins and carcinogens. As many toxins and carcinogens are ingested via food, the human gastrointestinal tract not only has an important role in the uptake of essential nutrients, but also acts as a first barrier against such harmful constituents of the food. Therefore, the gastrointestinal mucosa contains high levels of detoxification enzymes such as cytochromes-P450, glutathione S-transferases and UDP-glucuronosyltransferases. AIM: To compare the UDP-glucuronosyltransferase detoxification capacity in small intestinal mucosa of patients with coeliac disease vs. that in normal controls. METHODS: We assessed UDP-glucuronosyltransferase enzyme activities towards 4-methylumbelliferone in small intestinal biopsies of patients with coeliac disease (n = 22) and age- and sex-matched controls (n = 27). RESULTS: Small intestinal UDP-glucuronosyltransferase enzyme activity in controls was significantly higher than in patients with coeliac disease: 0.55 +/- 0.27 vs. 0.35 +/- 0.16 nmol/min mg protein, respectively (mean +/- s.d., P = 0.005). DISCUSSION: The low small intestinal UDP-glucuronosyltransferase detoxification activity in patients with coeliac disease may result in a deficient detoxification of potential carcinogens, and thus could explain in part the relatively high small intestinal cancer risk in these patients.

Contains fulltext : 49649.pdf (publisher's version ) (Closed access), OBJECTIVE: To stabilise the disease process in women with early onset severe preeclampsia and/or HELLP syndrome by enhancing maternal antioxidants effects of glutathione. STUDY DESIGN: In a randomised, double-blind, placebo-controlled trial, women with severe preeclampsia and/or HELLP syndrome received oral N-acetylcysteine. Primary outcome measures were disease stabilisation expressed as treatment-to-delivery interval and biochemical assessment of glutathione and parameters of oxidative stress. Secondary outcome measures were maternal complications, rate of caesarean section, stay at intensive care unit, postpartum hospital stay and neonatal morbidity and mortality. Analyses were done by intention-to-treat using Wilcoxon's two-sample test and regression analysis. RESULTS: Median treatment-to-delivery interval was not significantly different between the N-acetylcysteine and placebo group. The whole blood and plasma levels of glutathione and other thiols were not affected by N-acetylcysteine administration, except for plasma homocysteine concentrations, which were lower in the N-acetylcysteine group. There were no differences found in maternal nor neonatal secondary outcome measures between both groups. CONCLUSION: Oral N-acetylcysteine administration does not stabilise the disease process of early onset severe preeclampsia and/or HELLP syndrome.

Contains fulltext : 50658.pdf (publisher's version ) (Closed access), BACKGROUND: An ileal pouch-anal anastomosis has become the most widely accepted procedure for surgical treatment of patients with ulcerative colitis (UC). The primary function of the ileum within the pouch changes from absorption to storage. Malignancies have been described in the pouch mucosa. The detoxifying glutathione S-transferase (GST) enzymes are involved in the mucosal protection against toxins and carcinogens. Levels of GSTs are much higher in the ileum as compared with the colon. The adaptation of the ileal pouch mucosa into a more colon-like phenotype possibly influences the activity and levels of GST. This study compares the detoxification capacity of GST of the afferent ileal limb mucosa with the ileal pouch mucosa of patients with UC. METHODS: Biopsies from normal-appearing mucosa from the ileal pouch and the ileal afferent limb were obtained from 18 patients with UC. GST isoforms were quantified by immunoblotting. GST activity was measured spectrophotometrically, and glutathione and cysteine levels were determined by high-performance liquid chromatography. RESULTS: The GST activity and GSTA1+A2 levels were significantly lower in the pouch compared with the afferent ileal limb of patients with UC, whereas the GSTP1 levels were higher in the pouch. No differences were observed in the levels of GSTM1, GSTT1, glutathione, or cysteine. CONCLUSIONS: The lower GST detoxification activity in the pouch mucosa of patients with UC may result in higher levels of toxins and carcinogens and thus partly contribute to the risk of developing malignancies in the pouch.

Contains fulltext : 49867.pdf (publisher's version ) (Closed access), BACKGROUND: Thiols are scavengers of reactive oxygen species (ROS). We aim to investigate associations between thiols in various fluids in (sub)fertile couples and fertility outcome parameters. METHODS: In 156 couples undergoing assisted reproduction techniques (ART), we measured the concentrations of glutathione (GSH), cysteine (Cys), homocysteine (Hcy) and cysteinylglycine (CGS) and fertility outcome parameters in the ejaculate, purified spermatozoa and follicular fluid. RESULTS: All thiols were detectable in most ejaculates, spermatozoa and follicular fluids, of which Cys concentrations were highest. Thiol concentrations in the ejaculate were similar in fertile and subfertile men. However, Hcy in follicular fluid was higher in women with endometriosis compared with women in the idiopathic subfertile group (P=0.04). The GSH, Cys, Hcy and CGS concentrations in spermatozoa of subfertile men were significantly higher compared with men in the idiopathic subfertile group and fertile men (P<0.001). Most notably, Hcy concentrations in both the ejaculate and follicular fluid were negatively associated with embryo quality on culture day 3 in the IVF/ICSI procedure. CONCLUSIONS: Spermatozoa of subfertile men contain significantly higher thiol concentrations as compared with those of fertile men. The detrimental effect on embryo quality of a high Hcy concentration in the ejaculate and in follicular fluid is intriguing and may suggest that Hcy is inversely associated with fertility outcome.

Contains fulltext : 50677.pdf (publisher's version ) (Closed access), The duodenum is the main site for (pre-) malignant extracolonic manifestations in patients with familial adenomatous polyposis (FAP). Changes in the E-cadherin/beta-catenin complex play a pivotal role in the development of malignancies. Loss of E-cadherin has been described in association with loss of SMAD4. To elucidate the pathways leading to the development of duodenal adenomas in patients with FAP, the distributions of E-cadherin, SMAD4, and beta-catenin were analyzed. Furthermore, differences between the duodenum and colon were evaluated. Normal FAP duodenum (n = 13) and FAP duodenal adenomas (n = 50; total, 21 patients) were compared with non-FAP duodenal adenomas (n = 7) and normal non-FAP duodenum (n = 15) by immunohistochemical staining for extracellular and intracellular E-cadherin, beta-catenin, and SMAD4. Colonic biopsies of 10 patients with FAP were also studied, as well as non-FAP colonic adenomas (n = 26) and non-FAP normal colon (n = 12). Compared with the intracellular component of E-cadherin that was present in all cases, a significant loss of extracellular E-cadherin was observed in both duodenal and colonic adenomas and normal tissue of patients with FAP. Nuclear localization of beta-catenin was more often observed in duodenal FAP adenomas compared with non-FAP adenomas. Loss of nuclear SMAD4 was seen in the duodenum and, to a higher degree, in the colon of patients with FAP, as well as non-FAP patients. The loss of extracellular E-cadherin in the normal duodenal and colonic mucosa of patients with FAP might play a role in the high susceptibility of these tissues for (pre-) malignant transformation.

Contains fulltext : 51005.pdf (publisher's version ) (Closed access), BACKGROUND: Adenomas can develop in the pouch after colectomy with ileal pouch-anal anastomosis (IPAA) in patients with familial adenomatous polyposis (FAP). Glutathione S-transferases (GSTs) have a protective role in carcinogenesis. GST activity is much higher in the ileum than in the colon. The present study examined the hypothesis that the protective capacity of GSTs may be lowered as a result of colonic metaplasia of the ileal pouch. METHODS: Levels of GSTs, glutathione and cysteine, and the degree of inflammation and colonic metaplasia were quantified in biopsies from the pouch and afferent loop of 26 patients with FAP. RESULTS: GST enzyme activity, and levels of GST alpha, glutathione and cysteine in the pouch were significantly lower than those in the afferent loop (308 versus 398 nmol per min per mg protein (P<0.001), 4604 versus 5286 ng per mg protein (P=0.010), 27.1 versus 34.8 nmol per mg protein (P=0.023) and 0 versus 4.8 nmol per mg protein (P=0.009) respectively). No correlation was found between inflammation or colonic metaplasia of the pouch and GST enzyme activity in the pouch. CONCLUSION: After IPAA, GST detoxification activity in the pouch is significantly lower than that in the afferent ileal loop, which may promote tumorigenesis.

Contains fulltext : 50115.pdf (publisher's version ) (Closed access), Surface proteins play important pathophysiological roles in health and disease, and accumulating proteomics-based studies suggest that several "non-membrane" proteins are sorted to the cell surface by unconventional mechanisms. Importantly, these proteins may comprise attractive therapeutic targets and novel disease markers for colon cancer. To perform a proteomics-based inventory of these so-called "anchorless" surface proteins, intact colon adenocarcinoma SW480 cells were labeled with membrane-impermeable biotin after which only soluble biotinylated proteins were isolated and identified by nanoLC-MS/MS. Computer-assisted analysis predicted that only 9 of the 97 identified surface-exposed proteins have predicted secretory signal peptides, whereas 2 other proteins have a putative transmembrane segment. Of the 9 proteins with putative signal peptides, 1 was predicted to be retained at the cell surface by a GPI-anchor, whereas 5 other proteins contained an ER-retention motif (KDEL) that should prevent them from being sorted to the cell surface. The remaining 86 soluble "surface" proteins lack known export signals and the possibility that these proteins are candidate substrates of non-classical transporters or exported by unconventional mechanisms is discussed. Alternatively, the large number of "intracellular" and ER-resident proteins may imply that biotinylation approaches are not only specific for surface proteins, but also biased against a certain subset of non-surface proteins. This underscores the importance of post-proteomic verification of proteomics-based inventories on surface-exposed proteins, which eventually should reveal to which extent non-classical export and retention mechanisms contribute to the sorting of "anchorless" proteins to the surface of colon tumor cells.

Item does not contain fulltext, BACKGROUND: The glutathione S-transferases (GST) can metabolise endogenous and exogenous toxins and carcinogens by catalysing the conjugation of diverse electrophiles with reduced glutathione (GSH). Variations of GST enzyme activity could influence the susceptibility of developing cancers in certain areas of the gastrointestinal tract. AIMS: The expression of the components of the glutathione system in the colon was investigated with respect to age, gender and localisation. METHODS: Biopsies of macroscopically normal mucosa from both proximal and distal colon were collected from 208 patients (106 females, 102 males; mean age 61 years), who underwent colonoscopy for various clinical reasons. GSH content, total GST enzyme activity and the levels of the GST isoenzymes glutathione S-transferase P1 (GSTP1) and glutathione S-transferase M1 (GSTM1) were determined. RESULTS: GST enzyme activity, GSH and GSTP1 levels decreased significantly from proximal to distal colon (GST activity: 264 vs. 244 nmol/min/mg protein, p < 0.001, GSH content: 32 vs. 30 nmol/mg protein, p = 0.022 and GSTP1 levels: 2.25 vs. 2.10 mug/mg protein, p < 0.001). In female patients there was a significant stepwise increase of GST-activities and GSTP1 levels from the age of under 50 years to over 70 years. Oral sex hormone substitution among female patients between 50 and 70 years suppressed GST-activities and GSTP1 content. CONCLUSIONS: The GSH-system in the colonic mucosa is expressed at a lower level in the distal colon (sigma) than in the colon transversum; whether this small difference translates into variations of incidence of colorectal cancer remains to be seen. Females express higher enzyme levels as they grow older, while in males no significant age effects were found. Elderly females might be better equipped with protective GSH-enzymes in the colon than males and this could contribute to the lower incidence of colorectal carcinomas in females.

Contains fulltext : 50669.pdf (publisher's version ) (Closed access), BACKGROUND: Colon carcinogenesis is a multifactorial process influenced by hereditary as well as environmental factors. The glutathione/glutathione S-transferase detoxification system in the colon is important for protection against carcinogens. We investigated the levels of glutathione/glutathione S-transferase in normal colon mucosa of patients with colorectal cancer and in patients at high risk for colorectal cancer compared with those in healthy controls. MATERIALS AND METHODS: Glutathione content was analyzed by high-performance liquid chromatography, and glutathione S-transferase enzyme activity by spectrophotometric determination with 1-chloro 2,4-dinitrobenzene. Normal colon tissue of patients with colon adenoma (n = 64), colorectal cancer (n = 37), familial adenomatous polyposis (FAP; n = 19), hereditary non-polyposis colorectal cancer families with (HNPCC+Ad; n = 34) or without (HNPCC-Ad; n = 33) adenoma was investigated. RESULTS: Glutathione levels were significantly lower in the normal colon mucosa of patients with cancer, FAP, HNPCC-Ad or HNPCC+Ad compared with adenoma patients or healthy controls. Glutathione S-transferase enzyme activity in the distal colon was significantly lower in patients with cancer or FAP compared with the adenoma patients or healthy controls, whereas values in carcinoma patients were significantly lower compared with both the HNPCC-Ad and HNPCC+Ad groups. CONCLUSIONS: An association of low colonic glutathione/glutathione S-transferase activity levels and high clinical risk for the development of colorectal cancer was observed. This low glutathione detoxification capacity might contribute to the colon cancer risk.

Contains fulltext : 48468.pdf (publisher's version ) (Closed access), Interest in mechanisms of colon cancer prevention by food compounds is strong and research in this area is often performed with cultured colon cancer cells. In order to assess utility for screening of potential cancer-preventive (food) compounds, expression profiles of 14 human cell lines derived from colonic tissue were measured using cDNA microarrays with 4000 genes and compared with expression profiles in biopsies of human colon tumours and normal tissue. Differences and similarities in the gene expression profiles of the cell lines were analysed by clustering and principal component analysis (PCA). Cytoskeleton genes and immune response genes are two functional classes of genes that contributed to the differences between the cell lines. A subset of 72 colon cancer-specific genes was identified by comparing expression profiles in human colon biopsies of tumour tissue and normal tissue. A separation of the cell lines based on the tumour stage of the original adenocarcinoma was observed after PCA of expression data of the subset of colon cancer-specific genes in the cell lines. The results of this study may be useful in the ongoing research into mechanisms of cancer prevention by dietary components.

Contains fulltext : 48773.pdf (publisher's version ) (Closed access), BACKGROUND: To analyse the post-partum concentrations of intra- and extra-cellular blood antioxidants in women with uncomplicated pregnancies. METHODS: Whole blood and plasma thiols, plasma vitamin E and C, serum cholesterol and triglyceride, ferric reducing ability of plasma (FRAP) concentrations were compared between women delivered by caesarean section (n=17) or spontaneous delivery (n=10). A repeated mixed model was used for statistical analysis. RESULTS: The majority of whole blood thiols increased significantly in both groups the first days post-partum. However, within the caesarean group free cysteine, oxidised cysteine, homocysteine and glutathione and plasma cysteine and homocysteine levels dropped significantly after 24 h, while FRAP levels peaked significantly in this group. Plasma vitamin E levels decreased significantly in both groups within 24 to 48 h after delivery. Independent of the way of delivery whole blood and plasma thiols were significantly increased and vitamin E levels were significantly decreased 3 months post-partum while plasma vitamin C levels and FRAP were unchanged compared to ante-partum levels. DISCUSSION: Decreased plasma vitamin E levels shortly post-partum are associated with decreased lipid peroxidation. The 24 h post-partum drop of some plasma and whole blood thiols in the caesarean group may be due to prolonged fasting.

Contains fulltext : 47876.pdf (publisher's version ) (Closed access), OBJECTIVE: Cardiac surgery is an important risk factor for the development of acute renal failure. Cytosolic enzymes glutathione S-transferase (GST) A1 and P1 are present selectively in proximal and distal tubular cells, respectively. We determined the extent and site of tubular injury and examined if GST excretion may predict a clinically relevant change in renal function. DESIGN AND SETTING: A prospective, observational study in 84 consecutive patients in the cardiac surgery intensive care unit of the University Medical Centre Nijmegen. MEASUREMENTS AND RESULTS: Urinary GST enzyme excretion was determined 0-4 h and 20-24 h after cardiac surgery by enzyme-linked immunosorbent assay. Data are expressed as median and 5-95% range. Urinary excretion of GSTA1 was increased: 1.25 microg/mmol [0.31-10.20] creatinine at t =0-4 h ( p <0.0001, compared with controls; 0.25 [0.1-0.8]) and returned to normal values at t =20-24 h. Excretion of GSTP1 was 2.11 microg/mmol [0.52-17.82] creatinine ( p <0.0001) at t =0-4 h and remained significantly elevated: 0.84 [0.30-16.86] at t =20-24 h ( p =0.01) compared with controls (0.5 [0.2-1.1]). The ten patients with the highest urinary excretion of GSTA1 or GSTP1 did not demonstrate a different plasma creatinine level on postoperative day 3, compared with the ten patients with the lowest urinary excretion of GSTA1 or GSTP1. CONCLUSION: Uncomplicated cardiac surgery results in a statistically significant increase in the urinary excretion of GSTA1 and GSTP1 as compared with healthy controls, indicating proximal and distal tubular damage. However, this small increase in urinary excretion of GSTs is not associated with clinically relevant renal injury.

Contains fulltext : 49159.pdf (publisher's version ) (Closed access), OBJECTIVE: To evaluate the physical and mental health of women with a history of severe preeclampsia. METHODS: In a historical cohort study 131 former patients with a history of severe preeclampsia and 127 control patients received questionnaires about experienced physical and mental complaints after delivery. At a follow-up visit blood pressure, body mass index, and proteinuria were measured and venous blood was drawn. RESULTS: Former patients experienced significantly (p < 0.001) more frequent problems of headache (31% vs. 2%), right upper quadrant pain (16% vs. 1%), visual disturbances (21% vs. 1%), tiredness (66% vs. 27%), subjective loss of concentration (37% vs. 16%), and mental health (37% vs. 6%) compared with controls. When present, these health problems, except for tiredness, lasted significantly more often beyond six months postpartum compared to controls. Admittance to the intensive care unit was associated with headache, and subjective loss of memory and concentration over a longer period of time. The risk of recurrence of severe preeclampsia was a subject of concern in 20% of former patients. At follow-up, systolic and diastolic blood pressures were significantly higher (p < 0.001) among former patients. CONCLUSION: Patients with a history of severe preeclampsia more frequently reported physical and mental complaints, also during a longer period of time.

Contains fulltext : 47773.pdf (publisher's version ) (Closed access), It is hypothesized that excessive generation of reactive oxygen species (ROS) by phagocytes or leakage from mitochondria may harm key genes or proteins responsible for intestinal cell homeostasis. This may initiate the multistage process of colon cancer development. The present study investigates whether ROS production by whole blood may contribute to the etiology of colorectal cancer (CRC). Whole-blood oxygen radical production was measured by luminol-enhanced chemiluminescence and performed in fourfold with and without the stimuli phorbol 12-myristate 13-acetate (PMA) and serum-treated zymosan (STZ). We evaluated patients (i) with a history of sporadic CRC at least 3 months after surgery, (ii) who were hereditary nonpolyposis colorectal cancer (HNPCC) gene carriers, and (iii) with familial adenomatous polyposis (FAP). For each patient group (n = 20) an age- and gender-matched healthy control group was measured. Unstimulated and PMA-stimulated values for maximal oxygen radical production were significantly higher in patients with sporadic CRC in comparison to controls (p = 0.01, p = 0.04, respectively). Furthermore, trends toward higher unstimulated and PMA-stimulated area under the curve chemiluminescence were seen in CRC patients compared with controls (p = 0.08, p = 0.09, respectively). In patients with HNPCC or FAP, unstimulated or PMA- or STZ-stimulated chemiluminescence did not differ compared to their control groups. In conclusion, whole-blood oxygen radical production was higher in patients with a history of sporadic CRC, in comparison with age- and gender-matched controls, which indicates that ROS may play a role in the etiology of sporadic CRC.

Contains fulltext : 48397.pdf (publisher's version ) (Closed access), PURPOSE: The gut fermentation product of dietary fiber, butyrate, inhibits growth of HT29, an established tumor cell line. It also induces detoxifying enzymes belonging to the glutathione S-transferase family (GSTs), namely hGSTM2, hGSTP1, hGSTA4, but not of hGSTT1 . Here we investigated kinetics of effects in HT29 and compared sensitivities with preneoplastic LT97 colon adenoma cells, to assess mechanisms of colon cancer chemoprevention in two stages of cell transformation. METHODS: We determined cell growth after butyrate treatment by quantifying DNA, GST expression by Northern/Western Blotting or biochemical analysis and butyrate consumption by measuring the residual concentrations in the cell culture supernatants. Stability of GST-theta (hGSTT1) mRNA was assessed in HT29 cells after inhibition of transcription with actinomycin D. RESULTS: LT97 adenoma cells consumed twofold more butyrate and were more sensitive to growth inhibition than HT29 (EC(50)1.9 mM and 4.0 mM, respectively). Butyrate did not induce GSTs, but instead reduced hGSTT1 in LT97 and HT29. CONCLUSIONS: Butyrate has suppressing-agent activities in human colon cells by inhibiting two survival factors, namely hGSTT1 and cell growth, with LT97 more sensitive than HT29. These findings indicate that butyrate formation in the gut lumen of humans could be protective by reducing survival of transformed colon cells.

RU Radboud Universiteit Nijmegen, 15 september 2005, Promotor : Jansen, J.B.M.J. Co-promotores : Nagengast, F.M., Peters, W.H.M., Contains fulltext : 47367.pdf (publisher's version ) (Open Access), The aetiology of sporadic colorectal cancer (CRC) is complex and involves genetic and lifestyle factors. To deal with the daily load of carcinogens, present in food, tobacco smoke etc., humans possess an efficient system of defence against such compounds and an important role is reserved for the detoxification enzymes UDP-glucuronosyltransferases (UGTs) and NAD(P)H:quinone oxidoreductase (NQO1). However, when the detoxification capacity of the colon is insufficient, this may lead to DNA mutations and damage of cells and ultimately to development of CRC. Here, we described that addition of dietary compounds to the diet of rats induced UGT enzyme activity in liver (flavone: 10.6x, coumarin: 6.2x, alpha-angelicalactone: 4.2x, D-limonene 3.2x) and colon (curcumin: 3.1x, coumarin: 2.7x, quercetin and alpha-angelicalactone: both 2.2x). Furthermore, it seems worthwhile to study the combined effect of dietary compounds, since additional or synergistic effects may occur. This could be concluded from a study in which a combination of flavone and alpha-angelicalactone was supplemented to the diet of rats. Enhanced enzyme activity might lead to more efficient detoxification of carcinogens and hence contribute to the prevention of CRC. However, the significance of results from animal studies should be verified in human intervention studies. Genetic polymorphisms in detoxification enzymes may result in less active enzymes, which may lead to reduced conversion of carcinogens in the colon and eventually an increased risk for CRC. Here, we reported that polymorphisms in the detoxification enzymes UGT1A6, UGT1A7 and NQO1 were found significantly more often in patients with CRC as compared to healthy controls, indicating that the presence of these variant genotypes may contribute to development of CRC. Finally, we showed by measuring oxygen radical production in whole blood from patients with a history or at risk for CRC and age- and gender-matched controls, that oxidative stress migh

Item does not contain fulltext, New knowledge of placental development and function suggests that several common complications of pregnancy could share a similar origin. It is suggested that impaired placental development in early pregnancy may lead to placental oxidative stress and subsequently to the maternal syndromes such as recurrent early pregnancy loss and pre-eclampsia. Oxidative stress has been most extensively investigated in pre-eclampsia, resulting in hundreds of publications and many reviews. In general the literature points to the presence of placental and maternal oxidative stress. However, conformity amongst the relevant data is not absolute, most probably the result of the diversity of biomarkers investigated and the methods employed to assess oxidative stress, which generally depend on the assessment of end products of oxidative stress. Recently, new techniques have been developed that use different approaches based on the "real-time" measurement of oxidative stress by the redox status of thiols or the assessment of superoxide generation, whereas the role of Phase I/Phase II biotransformation pathways in oxidative stress was recognised. This review focuses on this biotransformation system, the thiol redox status and the involvement of these systems in oxidative stress associated with reproduction and pregnancy disorders, with the emphasis being laid on the syndrome of pre-eclampsia.

Contains fulltext : 47896.pdf (publisher's version ) (Closed access)

Contains fulltext : 48901.pdf (publisher's version ) (Closed access), OBJECTIVE: This study was undertaken to determine whether the N-acetyltransferase (NAT) phenotype contributes to the susceptibility for the development of preeclampsia. STUDY DESIGN: The NAT acetylator status was determined by measuring urinary caffeine metabolites in 134 nonpregnant women with a history of preeclampsia and in 109 control women with uncomplicated pregnancy. The chi(2) and logistic regression analyses were used for statistical evaluation of differences in acetylator status. RESULTS: Significantly more fast acetylators were found among the women with a history of preeclampsia (46.3%) than among the controls (25.4%). Fast acetylators showed an odds ratio of 2.5 (95% CI 1.4-4.3) for preeclampsia. No differences in the acetylator status were found between women with a history of preeclampsia only and those with the HELLP syndrome as well. CONCLUSION: The fast NAT acetylator status, which may result in altered NAT detoxification capacity, is associated with preeclampsia.

Contains fulltext : 48168.pdf (publisher's version ) (Closed access), Objective. To investigate an association between a family history of cardiovascular disease and severe preeclampsia and/or HELLP syndrome (Haemolysis, Elevated Liver enzymes, Low Platelets). Methods. One hundred twenty-eight women with a history of severe preeclampsia and/or HELLP syndrome and 123 women with previous uncomplicated pregnancies only were included in the study. All participants completed questionnaires about diagnoses of cardiovascular diseases, hypertension, and hypercholesterolemia among their first-degree relatives, which were subsequently confirmed by the relatives' general practitioners. The main outcome measures were the prevalence of cardiovascular diseases, hypertension, and hypercholesterolemia among first-degree relatives of both groups. Statistical analysis was done using chi(2)-analysis. Results. The prevalence of familial cardiovascular disease among women with a history of severe preeclampsia and/or HELLP syndrome (23%) compared to controls (19%) was not significantly different (OR 1.3, 95%CI 0.7-2.5). However, women with a history of severe preeclampsia and/or HELLP syndrome more often had one or more first-degree relatives with hypertension and/or hypercholesterolemia before the age of 60 years compared to controls (54% vs. 32%, respectively; OR 2.6, 95%CI 1.5-4.3). The prevalence of hypertension and hypercholesterolemia among first-degree relatives, irrespective of age, also was significantly higher among women with a history of severe preeclampsia and/or HELLP syndrome as compared to controls (60% vs. 42%, respectively; OR 2.0, 95%CI 1.2-3.4). Conclusion. Severe preeclampsia is associated with a positive family history of hypertension and/or hypercholesterolemia.

KUN Katholieke Universiteit Nijmegen, 15 december 2004, Promotores : Steegers, E.A.P., Merkus, J.M.W.M., Jansen, J.B.M.J. Co-promotor : Peters, W.H.M., Contains fulltext : 58799.pdf (publisher's version ) (Closed access)

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Contains fulltext : 59270.pdf (publisher's version ) (Closed access), OBJECTIVE: To investigate a possible mechanism that could lead to the subsequent development of cardiovascular diseases (CVD) in women with a history of severe pre-eclampsia. DESIGN: Case-control study. SETTING: University Medical Centre Nijmegen, The Netherlands. SAMPLE: Non-pregnant women with a history of severe pre-eclampsia (n= 131) and women with an uncomplicated obstetric history (n= 94). METHODS: Total plasma levels of cysteine (tCys), homocysteine (tHcy), cysteinylglycine (tCysGly) and glutathione (tGSH), the free-to-oxidised ratio of these thiols in whole blood, the glucose-6-phosphate dehydrogenase (G6PDH) enzyme activity and antioxidant capacity were assessed at least 6 months following last pregnancy. MAIN OUTCOME MEASURE: Oxidative stress and antioxidant status. RESULTS: Women with a history of severe pre-eclampsia showed higher levels (mean [SD]) of tHcy (13.1 [5.0] versus 11.5 [4.8] micromol/L; P= 0.018) and tCysGly (37.5 [5.6] versus 34.0 [5.8] micromol/L; P= 0.0001) compared with controls, whereas tCys was lower (232 [31] versus 242 [39]; P= 0.029). The lower free-to-oxidised ratio of homocysteine (2.3 [0.8] versus 2.9 [1.0], P= 0.0001) among women with a history of severe pre-eclampsia as compared with control subjects might indicate a higher oxidant status for homocysteine. Previous severe pre-eclamptic patients had also a higher antioxidant capacity as compared with controls (0.79 [0.14] versus 0.74 [0.11] mmol Fe2+/L, P= 0.002). CONCLUSION: Since women with a history of severe pre-eclampsia showed elevated total homocysteine levels, which is an independent risk factor for CVD, and higher oxidised homocysteine levels in whole blood, these women may have an enhanced risk for the subsequent development of cardiovascular-related problems in later life.