sodium-nitrite and methylurea

The in vivo nitrosation capacity of third-instar larvae of Drosophila melanogaster was assessed using the wing somatic mutation and recombination test (SMART). Larvate derived from two different crosses, the standard cross (ST) and the high bioactivation cross (HB) both involving the recessive wing cell markers multiple wing hairs (mwh) and flare (flr3), were used. The HB cross is characterised by an increased cytochrome P450-dependent bioactivation capacity for promutagens and procarcinogens. The larvae were treated either with methyl urea, sodium nitrite or its combination. N-Nitrosomethylurea was used as a positive control. The wings of the resulting flies were analysed for the occurrence of mutant spots produced by various types of mutational events or by mitotic recombination. Methyl urea is negative in the ST and the HB cross, whereas sodium nitrite is weakly genotoxic in both crosses. However, the combination of both compounds produces highly increased frequencies of mutations and recombinations predominantly in the HB cross. The genotoxic effects produced by the combined treatments were considerably increased when mashed potatoes or an agar-yeast medium were used for the treatment instead of the standard instant medium. Treatment of larvae with the mixture resulting from the in vitro reaction of nitrosation precursors also resulted in high frequencies of induced spots comparable to those recorded with the potent genotoxin N-nitrosomethylurea. Further experiments showed that the genotoxic effect resulting from the in vivo exposure to nitrosation precursors can be reduced by co-treatment with catechin, a known nitrosation inhibitor. The present study demonstrates that the wing spot test is well suited for the determination of genotoxicity produced by in vivo nitrosation processes and for the study of their modulation by individual compounds or dietary complex mixtures.

Topics: Animals; Biotransformation; Catechin; Crosses, Genetic; Cytochrome P-450 Enzyme System; Drosophila melanogaster; Female; Larva; Male; Methylnitrosourea; Methylurea Compounds; Mutagenicity Tests; Mutagens; Recombination, Genetic; Sodium Nitrite; Wings, Animal

For carcinogenic risk assessment of combinations of N-nitroso precursors in man, the effects of feeding methylurea (MU) or morpholine (Mor) plus sodium nitrite (NaNO2) were investigated using a multi-organ carcinogenesis model. In experiment 1, to initiate multiple organs, groups of 10 or 20 male F344 rats were treated with 6 carcinogens targeting different organs. Starting a week after completion of this initiation phase, animals were given 0.1% MU or 0.5% Mor in their food and/or 0.15% NaNO2 in their drinking water for 23 weeks. The induction of tumors and/or preneoplastic lesions in the forestomach and esophagus was significantly increased in the group receiving MU plus NaNO2. The numbers and areas of liver glutathione S-transferase placental form (GST-P)-positive foci were significantly elevated with MU or Mor plus NaNO2. Experiment 2 was conducted to assess formation of N-nitroso compounds in the stomach, and to detect DNA adduct generation in target organs by immunohistochemical staining. Groups of 5 or 14 animals were starved overnight, then given 0.4% MU or 2.0% Mor in the diet, or basal diet alone for 1 h. Then NaNO2 or distilled water was given intragastrically. The mean gastric N-methyl-N-nitrosourea yield in the MU plus NaNO2 group was 7700 micrograms at 2 h after combined administration. The mean N-nitrosomorpholine yield in the group given Mor plus NaNO2 was 6720 micrograms. Immunohistochemically, N7-methyldeoxyguanosine-positive nuclei were evident in the forestomach epithelium at 8 h after the combination treatment with MU plus NaNO2.

Topics: Administration, Oral; Animal Feed; Animals; Biomarkers, Tumor; Carcinogenicity Tests; Carcinogens; Drug Interactions; Esophageal Neoplasms; Glutathione Transferase; Humans; Liver; Liver Neoplasms; Male; Methylurea Compounds; Morpholines; Neoplasms, Experimental; Nitroso Compounds; Precancerous Conditions; Rats; Rats, Inbred F344; Sodium Nitrite; Stomach Neoplasms; Water Supply

The effects of turmeric extract and its pure yellow pigments curcumin I, II and III were tested on the nitrosation of methylurea by sodium nitrite at pH 3.6 and 30 degrees C. The nitrosomethylurea formed was monitored by checking the mutagenicity in S. typhimurium strains TA1535 and TA100 without metabolic activation. Turmeric extract as well as curcumins exhibit dose-dependent decreases of nitrosation. Curcumin III was the most effective nitrosation inhibitor among the compounds tested. The simultaneous treatment of inhibitor with nitrosation precursors was essential and pre- or post-treatment of inhibitor had no effect on the mutagenicity of nitrosomethylurea. The binding of nitrite with the inhibitors was studied at pH 3.6 and 30 degrees C. Curcumin I shows a dose-dependent depletion of nitrite ions thus making nitrite non-available for nitrosation. Curcumin I and III when tested also showed a time-dependent depletion of nitrite ions at pH 3.6 and 30 degrees C. Curcumin III has a higher affinity for nitrite ions than curcumin I.

Topics: Catechols; Chemical Phenomena; Chemistry; Curcumin; Hydrogen-Ion Concentration; Methylnitrosourea; Methylurea Compounds; Mutagenicity Tests; Nitrites; Salmonella typhimurium; Sodium Nitrite; Time Factors

It was shown in experiments on 186 mice that formation of tumors of the lung and fore-stomach induced by injection of sodium nitrite in combination with aminopyrine or methylurea is inhibited following treatment with ascorbic acid, hexamethylenetetramine or sodium metabisulfite.

Topics: Aminopyrine; Animals; Ascorbic Acid; Cocarcinogenesis; Female; Lung Neoplasms; Male; Methenamine; Methylurea Compounds; Mice; Mice, Inbred Strains; Nitrites; Sodium Nitrite; Stomach Neoplasms; Sulfites

N-Nitroso compounds may well rank high among the genotoxic carcinogens present in our environment. Small amounts of such compounds may be formed in the human stomach after consumption of high-nitrate vegetables. Volatile nitrosamines can be conveniently determined but reliable methods of analysis for non-volatile N-nitroso compounds are still lacking. In this study we have used the 4-(4-nitrobenzyl)pyridine test to look for the formation of alkylating compounds such as N-nitroso-N-methylurea in a wide range of food products after incubation with nitrite under simulated gastric conditions. Our results indicate that many food products do not form alkylating compounds in appreciable amounts, even though the nitrite concentration used was five to ten times that found in saliva after a high-nitrate meal. Comparatively strong alkylating activity, however, was detected after incubation of samples of sauerkraut, certain dairy products (yoghurt, biogarde, quark, buttermilk and milk), wine and smoked mackerel. Samples of sauerkraut incubated with simulated gastric fluid, but without (added) nitrite, also displayed appreciable alkylating activity. The formation of alkylating substances in non-fat yoghurt was markedly inhibited by addition of ascorbic acid. The identity of the alkylating agents remains unknown. The isolation procedure was optimized for N-nitroso-N-methylurea, but will certainly result in the isolation of other compounds, such as C-nitroso-, C-nitro- or perhaps even C-nitroso-C'-nitro-compounds as well. Biogenic amines, glucosinolates, indole derivatives or other compounds may be involved as precursors. If alkylating agents are formed in vivo after ingestion of high-nitrate vegetables or drinking water, this is likely to occur only when the food products mentioned above are ingested simultaneously with or shortly after the nitrate load and not appreciably (except perhaps in the case of sauerkraut) when they are ingested alone, without a nitrate source. The health implications of these findings cannot yet be established. Many alkylating agents, however, have strong carcinogenic properties and continued investigation of food products (and their interaction products with nitrite) is indicated.

Topics: Alkylating Agents; Brassica; Dairy Products; Dinitrofluorobenzene; Fermentation; Food; Food Analysis; Gastric Juice; Methylnitrosourea; Methylurea Compounds; Mutagenicity Tests; Nitrites; Pyridines; Saliva; Sodium Nitrite

The formation of N-nitrosomethylurea (NMU) from methylurea (MU) and sodium nitrite in the guinea-pig stomach and the disappearance of NMU from the stomach were studied using a previously described method for NMU determination (Yamamoto et al. Fd Chem. Toxic. 1986, 24, 247). Guinea-pigs were used since they have only glandular stomachs and the pH of the gastric juice (1-2) is similar to that of humans. NMU was relatively stable in the isolated gastric contents of this species. When 2 mumol NMU was injected into the pylorus-ligated stomach of fasting guinea-pigs, about 50 and 37% of the NMU remained at 20 and 30 min, respectively. Some 19 and 42% remained 30 min after NMU was given orally by stomach tube to fasting and feeding guinea-pigs, respectively. NMU was detected in most blood samples irrespective of the administration procedure, but it disappeared rapidly from the blood after iv injection. Nitrite disappeared rapidly from the pylorus-ligated stomach, residual nitrite being less than 20% of the dose in 2.5 min. when 7.5 mumol MU and 15 mumol NaNO2 were co-injected into the ligated stomach, 3.1 mumol NMU was detected 10 min after the injection, followed by a gradual decrease. When MU and NaNO2 were given orally to the animals, 0.7-1.0 mumol NMU was detected in the stomach 10 min after the treatment. Thus NMU was shown to be formed readily in the stomach of the guinea-pig and to be absorbed from the stomach into the blood.

Topics: Animals; Fasting; Food; Gastric Juice; Gastric Mucosa; Guinea Pigs; Hydrogen-Ion Concentration; Kinetics; Male; Methylnitrosourea; Methylurea Compounds; Nitrites; Sodium Nitrite

The objective of this study was to simulate in vitro at least some of the conditions that prevail in man during ingestion of nitrate and nitrosable compounds. Human saliva has been chosen because most chemicals ingested through food will interact with saliva. The nitrosation of methylurea was used as a model because the nitrosation products can be readily detected by their mutagenic (his+ revertants of S. typhimurium) and clastogenic (chromosome aberrations in CHO cells) properties. The results show that human saliva inhibits the formation of mutagenic and clastogenic nitrosation products when present during nitrosation. A 50% inhibition of mutagenicity results from the addition of a saliva sample diluted at 5% of the original concentration. In the test system used a similar inhibitory effect was obtained by 2.5 mM ascorbic acid or 2.0 mM chlorogenic acid. The main inhibitory agents seem to reside in a deproteinized fraction which was filtered through an ultrafilter UM2 (greater than 1000 MW). At strong acid levels (below pH 2) the saliva loses its inhibitory effect on the nitrosation of methylurea. The contribution of saliva to the inhibition of endogenous nitrosation within the oral cavity or stomach is discussed.

Topics: Biotransformation; Cell Survival; Female; Humans; In Vitro Techniques; Kinetics; Male; Methylurea Compounds; Mutagenicity Tests; Mutagens; Nitrites; Nitroso Compounds; Saliva; Salmonella typhimurium; Sodium Nitrite