Endogenous N-nitroso compounds, and their precursors, present in bacon, do not initiate or promote aberrant crypt foci in the colon of rats

Processed meat intake is associated with increased risk of colorectal cancer. This association may be explained by the endogenous formation of N-nitroso compounds (NOC). The hypothesis that meat intake can increase fecal NOC levels and colon carcinogenesis was tested in 175 Fischer 344 rats. Initiation was assessed by the number of aberrant crypt foci (ACF) in the colon of rats 45 days after the start of a high-fat bacon-based diet. Promotion was assessed by the multiplicity of ACF (crypts per ACF) in rats given experimental diets for 100 days starting 7 days after an azoxymethane injection. Three promotion studies were done, each in 5 groups of 10 rats, whose diets contained 7%, 14%, or 28% fat. Tested meats were bacon, pork, chicken, and beef. Fecal and dietary NOC were assayed by thermal energy analysis. Results show that feces from rats fed bacon-based diets contained 10-20 times more NOC than feces from control rats fed a casein-based diet (all p < 0.0001 in 4 studies). In bacon-fed rats, the amount of NOC input (diet) and output (feces) was similar. Rats fed a diet based on beef, pork, or chicken meat had less fecal NOC than controls (most p < 0.01). No ACF were detected in the colon of bacon-fed uninitiated rats. After azoxymethane injection, unprocessed but cooked meat-based diets did not change the number of ACF or the ACF multiplicity compared with control rats. In contrast, the bacon-based diet consistently reduced the number of large ACF per rat and the ACF multiplicity in the three promotion studies by 12%, 17%, and 20% (all p < 0.01). Results suggest that NOC from dietary bacon would not enhance colon carcinogenesis in rats

In vivo Nitrosoproline formation and other risk factors in Costa Rican Children from high-and low-risk areas for gastric cancer

Artículo científico -- Universidad de Costa Rica, Instituto de Investigaciones en Salud. 1993The hypothesis that intragastric synthesis of N-nitroso compounds (NOC) in early life could play a role in gastric carcinogenesis was tested by applying the N-nitrosoproline (NPRO) test to about 50 children living in high- and low-risk areas for stomach cancer in Costa Rica. The median values of excretion of NPRO and the sum of three nitrosamino acids (micrograms/12 h urine) were 10-20% of those in adults from other geographical high-risk areas for stomach cancer. The urinary NPRO level after proline intake was higher in children from the high-risk area (P < 0.04) and markedly reduced after ingestion of ascorbic acid together with proline (P < 0.05). NPRO levels on the day of proline intake were highly correlated with levels of nitrate excretion (P < 0.001). Mean levels of total NOC in an aqueous (pH 2) extract of cooked beans from the high- and low-risk areas were similar. Acid-catalyzed nitrosation of the extract increased the total NOC concentration up to 1000-fold, but there was no difference between samples from the two areas. About 10% of bean extracts from both areas showed weak direct-acting genotoxicity in Escherichia coli; after acid-catalyzed nitrosation, all samples were genotoxic at similar levels. The diet of children in the low-risk area satisfied recommended levels of intake of energy and most nutrients except riboflavin and retinol equivalents. Diets from the high-risk area were deficient in energy intake and all nutrients except protein and vitamin C. Blood samples were collected from 276 children and young adults from the same areas and analyzed for serum antibodies against Helicobacter pylori. Although very high, no significant difference was found in the prevalence of IgG or IgA antibodies between the two regions.International Agency for Research on Cancer, Lyon, France.Universidad de Costa Rica, Instituto de Investigaciones en SaludUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto de Investigaciones en Salud (INISA

Exposure to N-nitrosamines and other risk factors for gastric cancer in Costa Rican children

Artículo científico -- Universidad de Costa Rica. Instituto de Investigaciones en Salud, 1991The hypothesis that endogenous chemical nitrosation in the normal stomach in early life could play a crucial role in inducing chronic atrophic gastritis/intestinal metaplasia in later life was tested by applying the N-nitrosoproline (NPRO) test to 12-h urine samples from about 50 children (aged 8-14 years) living in high- and low-risk areas for stomach cancer. The median values of NPRO and the sum of four nitrosamino acids analysed were 0.28-0.84 ag/12 h and 0.75-1.75 tig/12 h, respectively. The NPRO level after proline intake was significantly higher in children from a high-risk area than in those from a low-risk area (p < 0.04), and markedly reduced after ingestion of ascorbic acid and proline (p < 0.05). Urinary nitrate level was lower than that of adults. NPRO levels on the day of proline intake, however, correlated well with nitrate levels (p < 0.001), indicating that children in a high-risk area in Costa Rica have high endogenous nitrosation potential. Blood samples were also collected from about 300 children (aged 7-20 years) and analysed for antibodies against Campylabacter pylori, a suspected gastritis-causing bacteria. About 71% of children in both high- and low-risk areas for stomach cancer had antibodies. In addition, raw and cooked beans, which are consumed very frequently in Costa Rica, were collected from families in both areas and analysed for levels of nitrite/nitrate, total N-nitroso compounds and genotoxicity in the SOS chromotest. Mean levels of total N-nitroso compounds in an aqueous extract (pH 2) of cooked bean samples from high- and low-incidence areas were similar (0.4-0.6 nmol/g of cooked beans). Acid-catalysed nitrosation of the same aqueous extracts produced levels up to 2.4 pmol/g of cooked beans. There was no difference in mean levels of nitrosation-dependent total N-nitroso compounds between samples from the two areas. Only two out of 11 extracts from the low-incidence area and two out of 14 from the high-incidence area showed weak direct genotoxicity. After acid-catalysed nitrosation, all samples were genotoxic at similar levelsUniversidad de Costa Rica. Instituto de Investigaciones en Salud.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto de Investigaciones en Salud (INISA

Stress oxydatif et défenses anti-oxydantes dans la cancérogenèse gastrique et le cancer avéré

H. pylori provoque un stress oxydatif et expose au cancer gastrique. La nitric oxide synthase inductible (iNOS), la nitrotyrosine (NTYR) et la 8-hydroxy-2'-déoxyguanosine (8-OH-dG), marqueurs respectifs de la production des espèces oxygénées/ azotées réactives, des dommages aux protéines et à l'ADN, sont étudiés par immunohistochimie. Dans les gastrites, iNOS s'exprime dans les cellules inflammatoires activées. NTYR et 8-OH-dG siègent dans les cryptes profondes et pour la 8-OH-dG, dans les follicules lymphoïdes. Dans la muqueuse pré-cancéreuse, iNOS " migre " dans l'épithélium fovéolaire où les niveaux de NTYR et 8-OH-dG deviennent plus élevés. L'éradication bactérienne précoce diminue les trois marqueurs sans prévenir l'apparition de nouveaux dommages à l'ADN. Dans les cellules carcinomateuses, iNOS et NTYR sont corrélées au type histologique. Le stress oxydatif est médié par iNOS dans les adénocarcinomes tubuleux et à un moindre degré dans les carcinomes diffusLYON1-BU.Sciences (692662101) / SudocSudocFranceF

Chlorination of guanosine and other nucleosides by Hypochlorous Acid and Myeloperoxidase of Activated Human Neutrophils: Catalysis by nicotine and trimethylamine

International audienceActivated human neutrophils secrete myeloperoxidase, which generates HOCl from H$_2$O$_2$ and Cl$^-$. We have found that various (2'-deoxy)nucleosides react with HOCl to form chlorinated (2'-deoxy)nucleosides, including novel 8-chloro(2'-deoxy)guanosine, 5-chloro(2'-deoxy)cytidine, and 8-chloro(2'-deoxy)adenosine formed in yields of 1.6, 1.6, and 0.2%, respectively, when 0.5 mMnucleoside reacted with 0.5 mM HOCl at pH 7.4. The relative chlorination, oxidation, and nitration activities of HOCl, myeloperoxidase, and activated human neutrophils in the presence and absence of nitrite were studied by analyzing 8-chloro-, 8-oxo-7,8-dihydro-, and 8-nitroguanosine, respectively, using guanosine as a probe. 8-Chloroguanosine was always more easily formed than 8-oxo-7,8-dihydro- or 8-nitro-guanosine. Using electrospray ionization tandem mass spectrometry, we show that several chlorinated nucleosides including8-chloro(2'-deoxy)guanosine are formed following exposure of isolated DNA or RNA to HOCl. Micromolar concentrations of tertiary amines such as nicotine and trimethylamine dramatically enhanced chlorination of free (2'-deoxy)nucleosides and nucleosides in RNA byHOCl. As the G$-$463A polymorphism of the MPO gene, which strongly reduces myeloproxidase mRNA expression, is associated with a reduced risk of lung cancer, chlorination damage of DNA /RNA and nucleosides by myeloperoxidase and its enhancement by nicotine may be important in the pathophysiology of human diseases associated with tobacco habits

Molecular Analysis of the Cyclic AMP-Dependent Protein Kinase A (PKA) Regulatory Subunit 1A (PRKAR1A) Gene in Patients with Carney Complex and Primary Pigmented Nodular Adrenocortical Disease (PPNAD) Reveals Novel Mutations and Clues For Pathophysiology: Augmented PKA Signaling is Associated with Adrenal Tumorigenesis in PPNAD

We studied 11 new kindreds with primary pigmented nodular adrenocortical disease (PPNAD) or Carney complex (CNC) and found that 82% of the kindreds had PRKAR1A gene defects (including seven novel inactivating mutations), most of which led to nonsense mRNA and, thus, were not expressed in patients’ cells. However, a previously undescribed base substitution in intron 6 (exon 6 IVS +1G→T) led to exon 6 skipping and an expressed shorter PRKAR1A protein. The mutant protein was present in patients’ leukocytes and tumors, and in vitro studies indicated that the mutant PRKAR1A activated cAMP-dependent protein kinase A (PKA) signaling at the nuclear level. This is the first demonstration of an inactivating PRKAR1A mutation being expressed at the protein level and leading to stimulation of the PKA pathway in CNC patients. Along with the lack of allelic loss at the PRKAR1A locus in most of the tumors from this kindred, these data suggest that alteration of PRKAR1A function (not only its complete loss) is sufficient for augmenting PKA activity leading to tumorigenesis in tissues affected by CNC