K-ras oncogene mutations in sporadic colorectal cancer in The Netherlands Cohort Study

K-ras oncogene mutations in sporadic colorectal cancer in The Netherlands Cohort Study. Brink M, de Goeij AF, Weijenberg MP, Roemen GM, Lentjes MH, Pachen MM, Smits KM, de Bruine AP, Goldbohm RA, van den Brandt PA. Nutrition and Toxicology Research Institute Maastricht (NUTRIM), Department of Epidemiology, Maastricht University, PO Box 616, The Netherlands. [email protected] Activation of K-ras oncogene has been implicated in colorectal carcinogenesis, being mutated in 30-60% of the adenocarcinomas. In this study, 737 incident colorectal cancer (CRC) patients, originating from 120 852 men and women (55-69 years at baseline) participating in the Netherlands Cohort Study (NLCS), were studied in order to evaluate subgroups with respect to K-ras mutation status. Mutation analysis of the exon 1 fragment of the K-ras oncogene, spanning codons 8-29, was performed on archival colorectal adenocarcinoma samples of all patients using macrodissection, nested PCR and direct sequencing of purified fragments. The method of mutation detection was validated by the confirmation of reported K-ras status in CRC cell lines, a good correlation between fresh-frozen and routinely fixed, paraffin-embedded tissue, a detection limit of 5% mutated DNA and a good reproducibility. Various types of K-ras mutations were evaluated with respect to tumour sub-localization, Dukes' stage and tumour differentiation. In 37% (271/737) of the patients, the exon 1 fragment of K-ras gene was found to be mutated. The predominant mutations are G>A transitions and G>T transversions, and codons 12 and 13 are the most frequently affected codons. Patients with a rectal tumour were found to have the highest frequency of G>T transversions as compared with patients with a colon or rectosigmoid tumour. This difference appeared to be confined to women with a rectal tumour harbouring G>T transversions. No significant differences were observed for Dukes' stage with respect to types of K-ras mutation, which does not support direct involvement of the K-ras oncogene in adenocarcinoma progression. The equal distribution of K-ras mutations among cases with or without a family history of colorectal cancer argues against an important role for this mutation in familial colorectal cancer, and could imply that K-ras mutations are more probably involved in environmental mechanisms of colorectal carcinogenesis

Karma, morality, and evil

The doctrine of karma has been praised as a rational and morally edifying explanatory response to the existence of evil and apparent injustice in the world. Critics have attacked it as a morally misguided dogma that distorts one's vision of reality. This essay, after outlining the traditional doctrine, examines three criticisms that have been central to recent debates: firstly, that the doctrine offers no practical guidance; second, that it faces a dilemma between free will and fatalism; and third, that it involves a morally repugnant form of blaming victims for their own misfortunes. Possible responses are considered, the depth of the disagreement is highlighted, and a morally significant difference between alternative ways of articulating the belief in karma is analyzed

Adaptation to ER Stress Is Mediated by Differential Stabilities of Pro-Survival and Pro-Apoptotic mRNAs and Proteins

The accumulation of unfolded proteins in the endoplasmic reticulum (ER) activates a signaling cascade known as the unfolded protein response (UPR). Although activation of the UPR is well described, there is little sense of how the response, which initiates both apoptotic and adaptive pathways, can selectively allow for adaptation. Here we describe the reconstitution of an adaptive ER stress response in a cell culture system. Monitoring the activation and maintenance of representative UPR gene expression pathways that facilitate either adaptation or apoptosis, we demonstrate that mild ER stress activates all UPR sensors. However, survival is favored during mild stress as a consequence of the intrinsic instabilities of mRNAs and proteins that promote apoptosis compared to those that facilitate protein folding and adaptation. As a consequence, the expression of apoptotic proteins is short-lived as cells adapt to stress. We provide evidence that the selective persistence of ER chaperone expression is also applicable to at least one instance of genetic ER stress. This work provides new insight into how a stress response pathway can be structured to allow cells to avert death as they adapt. It underscores the contribution of posttranscriptional and posttranslational mechanisms in influencing this outcome

Formation of nitrosamines during consumption of nitrate- and amine-rich foods, and the influence of the use of mouthwashes.

Department of Health Risk Analysis and Toxicology, University of Maastricht, The Netherlands. We studied the formation of carcinogenic nitrosamines during consumption of food rich in nitrate and amines, and its possible inhibition by use of an antibacterial mouthwash. Twelve volunteers were fed a diet containing the high-nitrate vegetables lettuce or spinach during two periods of four consecutive days, in combination with fish products containing high levels of amines as nitrosatable precursors. During the two periods, the subjects used an antibacterial mouthwash containing chlorhexidine or a control mouthwash without antibacterial activity. Twenty-four-hour urine samples were collected after consumption of the meals, and saliva samples were collected 1 h after each meal. The nitrate and nitrite contents of the urine and saliva samples were determined by spectrophotometry (for nitrite) and HPLC (for nitrate). The concentrations of volatile nitrosamines in the urine samples were determined by gas chromatography-mass spectrometry. Significant increases in mean urinary nitrate levels (from 59 to 135 mg/24 h) and in mean salivary nitrate levels (from 10 to 56 microg/ml) and salivary nitrite levels (from 2 to 11 microg/ml) were observed during the consumption of food rich in nitrate and amines, as well as a significant increase in the mean urinary excretion of total examined volatile nitrosamines (from 2 to 7 nmol/24 h) and of N-nitrosodimethylamine (from 1.2 to 2.9 nmol/24 h). Use of the antibacterial mouthwash resulted in a decrease in mean salivary nitrite levels from 16 to 3 microg/ml and a decrease in mean urinary excretion of N-nitrosomorpholine (from 7.0 to 0.3 nmol/24 h). For the whole data set, significant correlations were observed between nitrate intake in food and urinary nitrate (p = 0.01; r2 = 0.07) and between urinary nitrate and urinary N-nitrosodimethylamine (p = 0.002; r2 = 0.11). In conclusion, consumption of a diet rich in nitrate and amines increases the risk of formation of carcinogenic nitrosamines. Use of an antibacterial mouthwash containing chlorhexidine can result in inhibition of nitrosamine formation

Activated neutrophils inhibit nucleotide excision repair in human pulmonary epithelial cells: role of myeloperoxidase

Neutrophils are thought to affect pulmonary carcinogenesis by promoting the metabolism of inhaled chemical carcinogens, causing enhanced formation of promutagenic DNA adducts. We hypothesized that neutrophils interfere with the removal of such DNA adducts by inhibiting nucleotide excision repair (NER) in target cells. Human alveolar epithelial cells (A549) were cocultured with activated neutrophils, and we observed a significant reduction of NER in the A549 cells, which was abrogated by addition of the myeloperoxidase (MPO) inhibitor 4-aminobenzoic acid hydrazide. The inhibitory effect of neutrophils could be mimicked by the MPO product hypochlorous acid (HOCl), which caused an acute, dose-dependent inhibition of NER in A549 cells. This was independent of cytotoxicity or ATP loss and persisted up to 24 h. These data were supported by showing that HOCl caused a delayed removal of DNA adducts in benzo[a]pyrene-diolepoxide-exposed A549 cells. The acute HOCl-induced inhibition of NER can only partly be explained by oxidative modification of repair proteins. To explain the more persistent effects of HOCl, we analyzed the expression of NER genes and found that HOCl significantly reduced XPC expression. In conclusion, these data indicate that neutrophils are potent inhibitors of nucleotide excision repair. This may provide a further biological explanation for the association between inflammation and lung cancer development. --Gungor, N., Godschalk, R. W. L., Pachen, D. M., Van Schooten, F. J., Knaapen, A. M. Activated neutrophils inhibit nucleotide excision repair in human pulmonary epithelial cells: role of myeloperoxidase

Placental telomere length decreases with gestational age and is influenced by parity: A study of third trimester live-born twins

AbstractBackgroundIn contrast to the postnatal period, little is known about telomere length (TL) during prenatal life. The decrease in placental TL remains unknown, although intra uterine growth retardation and preeclampsia are associated with shorter placental TL. The aim of this study is to assess the decrease of placental TL during the third trimester of gestation and to explore the role of potential “growth influencing factors”.MethodsThe study sample consisted of 329 live-born twins from the East Flanders Prospective Twin Survey. TL was determined using a multiplex quantitative PCR method. Gestational age, sex, birth order, placental characteristics, parity, maternal and paternal age, diabetes, hypertension, smoking, alcohol use, and socio economic status (SES) were considered “growth influencing factors”. Bivariable multilevel regression analysis with “growth influencing factors” was performed.ResultsPlacental TL ranged from 4.3 kbp to 84.4 kbp with a median of 10.8 kbp. Ln(TL) decreased in a linear fashion with an estimated TL decreasing from 13.98 kbp at 28 weeks to 10.56 kbp at 42 weeks. The regression coefficient of gestational age became smaller if considered together with SES (b = −0.017; p = 0.08) or diabetes (b = −0.018; p = 0.07) and bigger if considered together with parity (b = −0.022; p = 0.02), indicating that part of the association between gestational age and telomere length is explained by these three confounding factors.ConclusionPlacental TL decreases during the third trimester of gestation of live-born twins with approximately 25% indicating that telomere shortening may play a role in aging of the placenta