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Biomedical subjects

M C Archer

Publications and source records attributed to M C Archer.

At least 109 records · Page 6Linked to original sources

Reevaluation of nitrate and nitrite levels in the human intestine.

Analyses of human fecal and ileostomy samples by a method that is insensitive and free from interferences indicate that nitrate and nitrite levels in the intestine are lower than reported previously. Fecal nitrate and nitrite concentrations ranged from 0 to 14 mumol/kg (0 to 0.9 ppm) and 5 to 19 mumol/kg (0.3 to 0.9 ppm), respectively. Ileostomy samples contained from 0 to 7 mumol/kg (0 to 0.4 ppm) and 0 to 15 mumol/kg (0 to 0.7 ppm) for nitrate and nitrite, respectively. We also showed that, when deliberately added to feces samples, nitrate and nitrite were destroyed during a two-hr incubation period in a reaction that depended on the presence of microorganisms. The results suggest that conditions in the lower gastrointestinal tract favor denitrification, not nitrification as had been proposed previously.

Adolescent↗

Absence of volatile nitrosamines in human feces.

Using a method for nitrosamine analysis that gives high recovery values and that is free from artifactual synthesis of nitrosamines, we have shown that human feces do not contain volatile nitrosamines (detection limit, 0.1 to 0.5 microgram/kg). We also showed that nitrosation reactions are not catalyzed by fecal organisms. Following a 2-day anaerobic incubation of feces with either a secondary amine (dimethylamine, dipropylamine, or morpholine) or nitrite, no nitrosamine was formed. When the amine and nitrite were added together, nitrosamine was formed, but at a level of 2 to 20% of that formed in autoclaved feces under the same conditions. Nitrosamines were stable following anaerobic incubation with feces for up to 4 days. These results suggest that fecal organisms inhibit the chemical formation of nitrosamines instead of catalyzing it. When morphine and nitrate were added together, nitrosomorpholine was formed. Morpholine nitrosates so rapidly that it intercepts nitrite formed by the action of nitrate reductase before the nitrite can be further reduced. However, very high concentrations of morphine and nitrate, which are far from the conditions in normal feces, were required to form measurable nitrosomorpholine. We may conclude that N-nitroso compounds are unlikely to be formed in any significant amounts in the human colon.

Feces↗

Compounds formed by treatment of corn (Zea mays) with nitrous acid.

Nitrohexane has been identified as a major product formed following treatment of corn (Zea mays) with nitrous acid. Preliminary evidence suggests that another compound isolated from the nitrosated corn is an unsaturated nitrolic acid. As an aid to the analysis of N-nitro compounds, we have characterized the response of a chemiluminescence detector (Thermal Energy Analyzer) as a function of pyrolysis chamber temperature for several nitrosamines and for an aliphatic C-nitroso compound, an aromatic C-nitro compound, a nitramine and an alkyl nitrite. The response-temperature profiles are valuable in distinguishing among the various compounds and in optimizing the sensitivity of the detector for use in chromatography. Other tests, including photolysis and stability toward nitrite-scavenging reagents, further aid in distinguishing among the various compounds.

Chromatography, Gas↗

Inhibition of rat hepatic dimethylnitrosamine demethylase by cyclic and acyclic nitrosamines and secondary amines.

Inhibition of hepatic dimethylnitrosamine (DMN) metabolism by a variety of cyclic and acyclic nitrosamines was demonstrated. Nitrosoproline, a noncarcinogenic nitrosamine, behaved differently from the carcinogenic nitrosamines as an inhibitor for DMN-demethylase. Secondary amines corresponding to the nitrosamines inhibited DMN-demethylase in a manner similar to the nitrosamines.

Animals↗

Nitrosamine carcinogenicity: a quantitative Hansch-Taft structure-activity relationship.

Statistically significant correlations have been demonstrated between carcinogenic activity, water-hexane partition coefficients and electronic factors for an extensive series of N-nitroso compounds. Electronic factors were expressed by the Taft sigma* values of substituents on the carbon atoms alpha to the N-nitroso group. Such correlations indicate that transport of the carcinogen to its active site has an important effect on its potency. The correlations also implicate reactivity at the alpha-carbon in the determination of carcinogenic activity and point out various structural types which do not follow the general rule.

Animals↗

Microsomal metabolism of N-nitrosodi-n-propylamine: formation of products resulting from alpha- and beta-oxidation.

We have identified propionaldehyde, n-propranolo, isopropanol and N-nitroso-2-hydroxy-propylpropylamine following incubation of N-nitrosodi-n-propylamine with a microsomal fraction from rat liver. Based on the yields of the various products, we have shown that beta-oxidation occurs at about 15% of the level of alpha-oxidation, beta- as well as alpha-oxidation was shown to be carried out by the microsomal mixed function oxidase system. N-nitroso-2-hydroxy-propylpropylamine is further oxidized by the microsomal preparation to yield N-nitroso-2-oxopropylpropylamine.

Animals↗

Purification of acetate kinase by affinity chromatography.

A one-step procedure using affinity chromatography has been shown to purify to apparent homogeneity acetate kinase from a commercially available preparation and to partially purify the enzyme from a crude, cell-free extract. Since the gel's capacity for enzyme adsorption is controlled by the thermodynamics of ligand-enzyme interaction, maximization of the adsorption isotherm was attempted. Enzyme adsorption decreased logarithmically with increasing ionic strength but increased with increasing concentration of MgCl2. These competing effects caused the net adsorption of enzyme to increase to a maximum and then to decrease as the MgCl2 concentration was raised. The results allow a significant improvement in affinity column performance and have important implications for scale-up procedures.

Acetate Kinase↗

Nitrosamine formation in human saliva.

Nitrosamines formed when secondary amines were added to normal human saliva. Fractionation of saliva into cells and supernatant showed that factors that accelerated and retarded the nitrosation reaction were both present. Acidification of saliva greatly increased the nitrosamine yield, but differences in nitrosamine yield among saliva fractions were still observable.

Bacteria↗

Kinetic studies on the hepatic microsomal metabolism of dimethylnitrosamine, diethylnitrosamine, and methylethylnitrosamine in the rat.

The oxidative dealkylation of dimethylnitrosamine (DMN), diethylnitrosamine (DEN), and methylethylnitrosamine (MEN) by Sprague-Dawley rat liver microsomes was studied by a colorimetric assay for the simultaneous analysis of formaldehyde and acetaldehyde. Dealkylation in each instance followed Michaelis-Menten kinetics, but the Michaelis constant (Km) for DEN was an order of magnitude smaller than the Km for DMN. Km values for demethylation and deethylation of MEN were intermediate between those for DMN and DEN. DMN inhibited-deethylase, and DEN inhibited DMN-demethylase activity. The inhibition was of a mixed type in both instances.

Animals↗