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Respiratory tract lesions induced by sensory irritants at the RD50 concentration.

Exposure of mice to airborne sensory irritants causes a concentration-dependent depression of respiratory rate. The RD50 concentration (that concentration which elicits a respiratory rate decrease of 50%) has been predicted to be an unacceptable occupational exposure concentration due to intolerable sensory irritation and possible respiratory tract injury in humans. The purpose of this study was (1) to determine whether lesions occur in the respiratory tract of Swiss-Webster mice after exposure to the RD50 concentrations of ten sensory irritants and (2) to compare these changes with respect to type and severity. The RD50 values (ppm) of the chemicals studied are as follows: 2,4-toluene diisocyanate (0.4), acrolein (1.7), formaldehyde (3.1), chloropicrin (8.0), chlorine (9.3), sulfur dioxide (117), ammonia (303), hydrogen chloride (309), dimethylamine (511), and epichlorohydrin (687). After exposure of mice for 6 hr/day for 5 days, the respiratory tract was examined for histopathologic changes. All irritants produced lesions in the nasal cavity with a distinct anterior-posterior severity gradient. There was considerable variation in the extent, and nature of the lesions. The lesions ranged from slight epithelial hypertrophy or hyperplasia to epithelial erosion, ulceration, and necrosis with variable inflammation of the subepithelial tissues. Only chlorine, chloropicrin, and epichlorohydrin induced lesions in the lower respiratory tract. These findings give additional support to the potential value of the RD50 model for setting occupational exposure guidelines and predicting the risk of injury to the respiratory tract from exposure to airborne sensory irritants.

Animals↗

Isolation and characterization of a retinal-binding protein from the squid retina.

A retinal-binding protein (RALBP) was isolated from the squid retina, and purified by anion-exchange and size-exclusion chromatography. The molecular weight was determined to be 51,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by gel filtration. The purified sample showed absorption maxima at about 330 and 400 nm in addition to a protein band, indicating the occurrence of retinol and retinal, respectively. The relative heights of these two peaks varied from preparation to preparation, depending on retinoid ligands. Irradiation of RALBP caused no marked change in absorption, but the amount of 11-cis-retinal decreased to form a photosteady state mixture with all-trans- and 13-cis-retinals. RALBP was fairly stable even in the presence of hydroxylamine (100 mM), but was affected by sodium borohydride (30 mM) or borane dimethylamine (400 mM), with the retinal reduced to retinol. When incubated with metaretinochrome-carrying membranes in the dark, RALBP specifically took up 11-cis-retinal and lost all-trans-retinol. Upon further incubation of this RALBP with opsin-containing membranes, rhodopsin was progressively formed in the dark. Squid RALBP may act as a shuttle in transferring the 11-cis-retinal from metaretinochrome to opsin in the visual cells.

Animals↗

Biodegradation of alkyltrimethylammonium salts in activated sludge.

Trimethylamine, dimethylamine and methylamine (actually existing as a salt form in the culture medium) were identified as the intermediates of alkyltrimethylammonium salts in activated sludge obtained from a municipal sewage treatment plant. It was considered that the quaternary ammonium salts with long alkyl chains were degraded to tertiary amine by N-dealkylation at the first stage of the biodegradation pathway. The tertiary amine formed in this pathway rapidly disappeared. In the activated sludge, biodegradabilities based on biochemical oxygen consumption and dissolved organic matter were 7.2-53.7% and 97.4-100%, respectively. These results and the disappearance of intermediates as described above indicate that long chain alkyltrimethylammonium salts are ultimately biodegradable.

Biodegradation, Environmental↗

Oxidation of 1,1-dimethylhydrazine (UDMH) in aqueous solution with air and hydrogen peroxide.

The degradation of 1,1-dimethylhydrazine (UDMH), a component of some rocket fuels, was investigated using atmospheric oxygen and hydrogen peroxide. The reactions were carried out in the presence and absence of copper catalysis and at varying pH. Reactions were also carried out in the presence of hydrazine, a constituent, along with UDMH, of the rocket fuel Aerozine-50. In the presence of copper, UDMH was degraded by air passed through the solution; the efficiency of degradation increased as the pH increased but the carcinogen N-nitrosodimethylamine (NDMA) was formed at neutral and alkaline pH. Oxidation was not seen in the absence of copper. Production of NDMA occurred even at copper concentrations of < 1 ppm. Oxidation of UDMH with hydrogen peroxide also gave rise to NDMA. When copper was absent degradation of UDMH did not occur at acid pH but when copper was present some degradation occurred at all pH levels investigated. The production of NDMA occurred mostly at neutral and alkaline pH. In general, higher concentrations of hydrogen peroxide and copper favored the production of NDMA. Dimethylamine, methanol, formaldehyde dimethylhydrazone, formaldehyde hydrazone, and tetramethyltetrazene were also produced. The last three compounds were tested and found to be mutagenic.

Copper↗

Concentration of dimethylnitrosamine in the air of smoke-filled rooms.

In order to evaluate the contribution of volatile nitrosamines from tobacco smoke to indoor air pollution, N-nitroso-dimethylamine (NDMA) and N-nitroso-diethylamine (NDEA) were measured in indoor air under artificial and natural conditions. In controlled experiments under extreme conditions, we found that tobacco smoke-related NDMA levels above 0.07 ng/liter were associated with a highly irritating atmosphere which was scarcely tolerable to those present. In smoke-filled rooms under natural conditions NDMA levels ranged from 0.02 to 0.05 ng/liter except a minimum value of less than 0.01 ng/liter in a restaurant and a maximum of 0.07 ng/liter in a dancing bar. These NDMA levels are thus below comparable values reported by others. The NDMA/NDEA ratios found in air samples taken from some rooms under conditions of everyday life are quite different from those found in sidestream smoke of cigarettes. Irritation was not reported under natural conditions. From the results it is concluded that NDMA levels, measured under real life conditions, are usually not caused by tobacco smoke alone. Evidence for other sources of volatile nitrosamines is discussed.

Air↗

Amphiphilic detergents inhibit production of IgG and IgM by human peripheral blood mononuclear cells.

All types (cationic, anionic and non-ionic) amphiphilic detergents significantly inhibited the production of both IgG and IgM by human peripheral blood mononuclear cells after polyclonal activation in vitro. The most potent inhibitors were didodecyldimethylammonium bromide (DDAB) and (1-methyldodecyl)dimethylamine N-oxide (2-ATDNO). They were able to suppress effectively the immunoglobulin production in 10(-3)-10(-8) M concentrations. A medium inhibitory effect was observed with Slovapon, sodium dodecyl sulphate (SDS) and Triton X-100, while Slovanik showed an inhibition only in concentrations higher than 10(-2)%. These results suggest that amphiphilic detergents may be characterized as potential immunotoxic substances with very negative effects on the immunoglobulin production.

Detergents↗

Conductive cation transport in apical membrane vesicles prepared from fetal lung.

In order to characterise the apically-located conductive cation pathway of the type II pneumocyte, apical plasma membranes were prepared from mature fetal guinea pig lung. The protocol yielded purified apical membranes that enriched 19-fold with the brush border enzyme marker alkaline phosphatase; there was no significant contamination with other cellular membranes. A technique for imposing an outwardly-directed electrochemical Na+ gradient was used to amplify conductive 22Na+ uptake into vesicles. Uptake of 22Na+ was time-dependent, proportional to the magnitude of the Na+ gradient, specific and sensitive to the amiloride analogues phenamil and EIPA (apparent minimum Ki values of 50 nM and 10 microM, respectively, with maximum uptake inhibition of 42% and 39% at 100 microM). Uptake experiments in which the outwardly-directed Na+ gradient was replaced by outwardly-directed gradients of small monovalent cations and molecular cations were performed. The Na+/K+ permeability ratio was 1.2:1, and over the extended range of small monovalent cations, a permeability sequence of Na+ > K+ > Li+ > Rb+ > Cs+ was observed, indicating the presence of fixed negative charge in or spatially close to the pore. The molecular cation permeability sequence of NH4+ > methylamine+ > dimethylamine+ > choline+ > N-methyl-D-glucamine+ > tetraethylammonium+ > tetramethylammonium+, after transformation, gives an estimate of 8 A for the conducting pore diameter. These data are consistent with the presence in the apical membrane of fetal type II pneumocytes of a cation specific channel with low Na+ selectivity and amiloride sensitivity.

Animals↗

Lack of neuropathologic consequences of repeated dermal exposure to 2,4-dichlorophenoxyacetic acid in rats.

A 24% aqueous solution of the dimethylamine salt of 2,4-dichlorophenoxyacetic acid (2,4-D amine) was applied to the legs of male Fischer 344 rats 2 hr/day, 5 days/week, for 2 weeks. Because this concentration caused severe skin lesions, a second group of rats was treated similarly with a 12% solution of 2,4-D amine for 3 weeks. The 12% solution caused only mild skin changes. The plasma 2,4-D content, at the end of exposure, was nearly five times greater in the rats exposed to the 24% solution than to the 12% solution (323 vs 66.5 micrograms/ml). The severe skin changes probably facilitated absorption in the rats treated with the 24% solution. Rats treated with either concentration weighed less than controls. Although histologically normal, kidneys of treated rats weighed more than controls. The increased kidney weights were attributed to physiological adaptation due to active excretion of absorbed 2,4-D. Light microscopic examination of tissues, other than skin, revealed no differences between treated and control animals. There were no nervous system pathologic changes although the rats were exposed to sufficient amounts of 2,4-D amine to cause severe skin lesions, decreased body weights, and increased kidney weights.

2,4-Dichlorophenoxyacetic Acid↗

Acute, pharmacokinetic, and subchronic toxicological studies of 2,4-dichlorophenoxyacetic acid.

The single-dose oral LD50 values in Fischer 344 rats for technical-grade, 2,4-dichlorophenoxyacetic acid (2,4-D), esters, and salts ranged from 553 mg/kg (isobutyl ester in females) to 1090 mg/kg (dimethylamine salt in males). The LD50 values for the acid, esters, or salts, when expressed as acid equivalents, were consistent which suggests that the acute toxicity was due to 2,4-D per se. Acute dermal LD50 values in rabbits for the acid, esters, and salts were greater than 2000 mg/kg. Overall, these results indicate that the acute oral and dermal toxicity of 2,4-D are low. Pharmacokinetics were evaluated in male Fischer 344 rats given single oral doses of 10, 25, 50, 100, or 150 mg 2,4-[14C]D/kg. The amount of 2,4-D in the plasma, kidney, and urine 6 hr postdosing indicated that the urinary elimination of 2,4-D was saturated in male rats given oral doses in excess of 50 mg/kg. Subchronic dietary studies in male and female Fischer 344 rats used dose levels of 0, 15, 60, 100, or 150 mg/kg/day of purified or technical-grade 2,4-D acid for 13 weeks. Body weight gains were decreased for both sexes at the higher dose levels of purified and technical-grade 2,4-D acid. Kidney weights were increased in all treated male rats and in females given the higher three dose levels of purified 2,4-D. Treatment-related cytoplasmic alterations were present in the renal proximal tubules of most rats given 60 mg/kg/day and higher of purified or technical-grade 2,4-D; a few females given 15 mg/kg/day also had slight alterations in the cytoplasm of the proximal tubules. A dose-related degenerative change was identified in the descending proximal renal tubules of all male rats given the highest three dose levels of either test material and some given 15 mg/kg/day. Dose levels of 100 or 150 mg/kg/day of either compound for both sexes produced minimal swelling and increased staining homogeneity in the liver cells and were associated with a slight elevation of liver weight and serum glutamic pyruvic transaminase activity. Higher dose levels of technical-grade and purified 2,4-D decreased total serum tetraiodothyronine levels in female rats, however, the morphology of the thyroid gland was normal. The no-observed-effect level (NOEL) was less than 15 mg/kg/day for both purified and technical-grade 2,4-D acid.

2,4-Dichlorophenoxyacetic Acid↗

Proteins and amino acids as scavengers of nitrite: inhibitory effect on the formation of nitrosodimethylamine and diazoquinone.

Protein and its components in high concentrations similar to those found in the digestive tract effectively scavenged nitrite and thus inhibited the formation of mutagens from nitrite. Various amino acids at 100 mM, bovine serum albumin (BSA) and trypsinized casein at 100 mg/ml effectively decreased the nitrite level of 50 mM-nitrite solution at pH 3 and 37 degrees C. Most, but not all, amino acids can convert nitrite into nitrogen gas; Pro can be converted into non-mutagenic nitrosoproline, CySH to S-nitrosocysteine, Trp to weakly mutagenic nitrosotryptophan and Tyr to non-mutagenic diazotyrosine. BSA was cross-linked, probably owing to the reaction of Tyr and Lys residues. Most amino acids inhibited more than 50% of nitrosodimethylamine formation in the reaction between 200 mM-dimethylamine and 50 mM-nitrite at pH 3 and 37 degrees C; BSA inhibited 50% of nitrosodimethylamine formation, and trypsinized casein, 100%. Most amino acids inhibited more than 50% of p-diazoquinone formation in the reaction between 25 mM-phenol and 100 mM-nitrite at pH 3 and 37 degrees C; BSA inhibited 80%, and trypsinized casein, 95% of such formation. Trypsinized casein effectively inhibited the development of the mutagenic effects attributable to the formation of diazobamethan from bamethan and nitrite.

Amino Acids↗

Nitrosatable amines and nitrosamide formation in natural stimulants: Cola acuminata, C. nitida and Garcinia cola.

Three varieties of kola nut, Cola acuminata, C. nitida and Garcinia cola, of Nigerian origin, were analysed for their content of primary and secondary amines, and assessed for their relative methylating potential due to nitrosamide formation. Primary and secondary amines were determined as benzene sulfonamides by gas chromatography/thermal energy analysis (GC/TEA). Dimethylamine, methylamine, ethylamine and isopentylamine were detected in all kola nut varieties, while pyrrolidine, piperidine and isobutylamine were detected in one or more varieties. Estimated average total daily intake of aliphatic amines by a typical kola nut chewer varied from 260 to 1040 micrograms/day for secondary amines and from 2430 to 9710 micrograms/day for primary amines. Methylating activity of the nitrosated kola nuts, expressed as N-nitroso-N-methylurea equivalents, was also determined by GC/TEA. Methylating activity was significantly higher in kola nuts (170-490 micrograms/kg) than has ever been reported for a fresh plant product. These data suggest that the possible role of kola nut chewing in human cancer aetiology should be explored in countries where kola nuts are widely consumed as stimulants.

Amines↗

Potential of endogenous formation of volatile nitrosamines from Nigerian vegetables and spices.

Nitrosatability of dried Nigerian vegetables and spices was investigated under simulated gastric conditions. N-Nitroso-dimethylamine (NDMA) was the only volatile nitrosamine found above the preformed level except in ugwu where N-nitroso-piperidine (2.3 ppb) was also detected. The lowest NDMA value of 0.4 ppb was found in bitterleaf while onions had the highest level (14.7 ppb) of nitrosation. The results suggest that, under endogenous conditions, nitrosation of these foodstuffs could be an important contributory factor in cancer aetiology.

Food Preservation↗

Precursors of N-nitroso compounds in some Nigerian medicinal plants.

Twenty-seven tropical plants of medicinal importance were analysed for primary and secondary amines by chemiluminescence detection on a Thermal Energy Analyzer (TEA) modified for use on 'nitrogen mode' following derivatization with benzene sulphonyl chloride (BSC) and gas chromatographic (GC) separation of their sulphonamides. Nitrite was determined by colorimetry at 540 nm after diazotization with sulphanilamide and coupling with N-(1-naphthyl)ethylenediamine to form an azo dye. Nitrate was determined as nitrite following on-line reduction by granulated cadmium. Dimethylamine in the range of 0.5 ppm to 18.2 ppm was detected in 96% of samples, while pyrrolidine ranged between 0.7 ppm and 12.78 ppm in 14 samples. Isobutylamine, methylamine and ethylamine were the most ubiquitous primary amines. Largest number of secondary amines (four) was found in Azadirachta indica (Neem) while largest number of primary amines (six) was detected in Azadirachta indica and Tamarindus indica (Tsamiya) which also contained the highest amount of total primary amines (148.8 ppm). Nitrate and nitrite were seldom found in plant extracts whose pH were generally below 7.0. These findings suggests that early exposures to precursors of N-nitroso compounds via medicinal plants might contribute to total risk posed by environmental carcinogens in Nigeria.

Amines↗

Metabolism of naturally occurring propenylbenzene derivatives. III. Allylbenzene, propenyl benzene, and related metabolic products.

The acidic and neutral urinary metabolites of allylbenzene, propenylbenzene, 1'-hydroxyallylbenzene, and cynnamyl alcohol were identified and related through a common metabolic scheme. The rat metabolizes allylbenzene to 1'-hydroxyallylbenzene which can rearrange to yield cinnamyl alcohol whick is further metabolized. This mechanism is proposed to account for the appearance of "propenyl type" metabolites from allylbenzene compounds. Propenylbenzene is oxidized to cynnamyl alcohol. Both 1'-hydroxyallylbenzene and cunnamyl alcohol are excreted unchanged in the neutral extract when given to rats. Allylbenzene and 1'-hydroxyallylbenzene yield basic ninhydrin-positive metabolites. Allylbenzene is first oxidized on the benzylic carbon to form 1'-hydroxyallylbenzene, which is further oxidized to form phenyl vinyl ketone, which condenses with the secondary amines piperidine and dimethylamine to form tertiary aminopropiophenones (Mannich bases). Analogous compounds, cinnamyl alcohol and propenylbenzene, do not yield Mannich base metabolites. This proposed metabolic scheme is consistent with the chemical mechanism operative in the synthesis of Mannich base from allylbenzene via chromic acid oxidation followed by amine addition.

Animals↗

Aminopyrine uptake by guinea pig gastric mucosal cells. Mediation by cyclic AMP and interactions among secretagogues.

The role of cyclic nucleotides in regulating acid secretion by dispersed mucosal cells from guinea-pig stomach was examined by measuring first the ability of histamine and carbachol to stimulate [dimethylamine-14C]aminopyrine uptake and cyclic nucleotide metabolism and secondly, the effect of exogenous cyclic nucleotides on basal and stimulated [14C]aminopyrine uptake. The [14C]aminopyrine was found in an acidic, osmotically sensitive compartment, probably associated with the initial steps in acid secretion by these cells. Although histamine increased [14C]aminopyrine uptake and cyclic AMP synthesis as expected, histamine was approx. 10-fold more potent in inducing [14C]aminopyrine uptake. This dissociation of [14C]aminopyrine uptake and cyclic AMP metabolism process was further manifested by the observation that prostaglandin E1 failed to increase [14C]aminopyrine uptake, although it did cause a rise in cellular cyclic AMP. Furthermore, prostaglandin E1 did not alter the [14C]-aminopyrine uptake caused by histamine. Carbachol was found to increase the [14C]aminopyrine uptake and also to potentiate the ability of histamine to increase [14C]aminopyrine uptake. Carbachol, however, affected neither the histamine-induced increase in cyclic AMP nor the binding of [3H]histamine to the cells. Cimetidine, a histamine H2 receptor antagonist, blocked the [14C]aminopyrine uptake induced either by histamine alone or by the potentiating combination of histamine plus carbachol. These results suggest that cyclic AMP is mediating the action of histamine on [14C]aminopyrine uptake but changes in cyclic AMP per se are not necessarily the cause for the potentiated increase in [14C]aminopyrine uptake. Furthermore, the potentiated response observed with histamine plus carbachol on [14C]aminopyrine uptake occurs at a biochemical step distal to and not obviously related to cyclic AMP generation.

Aminopyrine↗

Localization, characterization and partial purification of TMAO-ase.

1. Formaldehyde (FA) and dimethylamine (DMA) are generated in several species of fish of commercial importance. 2. The present study describes the localization, partial purification and characterization of an enzyme recovered from cod kidney tissue which is responsible for the reduction of trimethylamine oxide (TMAO) to FA and DMA. 3. The enzyme, TMAO-ase was found to be bound to purified cod kidney lysosomes and was separated into at least four distinct isozymes by isoelectric focussing.

Animals↗

Gas chromatographic-mass spectrometric analysis of volatile amines produced by several strains of Clostridium.

A gas chromatographic--mass spectrometric technique is proposed for the analysis of volatile amines which were isolated from Clostridium cultures by vacuum distillation and concentrated as hydrochloride salts. Headspace sampling after alkalinization of the salts under vacuum was the most suitable for subsequent gas chromatographic analysis. With ammonia-loaded helium as carrier gas, methylamines were separated on 4.8% PEG 2OM + 0.3% potassium hydroxide on Carbopack B, and other volatile amines on 28% Pennwalt 223 + 4% potassium hydroxide on Gas-Chrom R. Bacterial volatile amines (dimethylamine, trimethylamine, isobutylamine, 3-methylbutylamine, etc.) were detected with a flame-ionization detector and identified by gas chromatography--mass spectrometry in electron-impact and chemical ionization modes.

Amines↗

Comparative effects of growth inhibitors on sterol metabolism in the nematode Caenorhabditis elegans.

An analogous series of dimethylalkyl compounds, consisting of four amines, an amide, and a phosphonate ester, inhibited motility and reproduction of the nematode Caenorhabditis elegans. Dimethylamines with straight-chain lengths of 12, 14, or 16 carbon atoms were equally active nematicides, causing greater than 80% population growth inhibition at a concentration of 25 ppm. The C12 straight-chain amine and its corresponding amide produced similar inhibition and were much more potent than either the corresponding C12 phosphonate or a C12 branched-chain amine. Inhibition of the delta 24-sterol reductase system was exhibited by all four amines, but not by the amide or phosphonate, in the following order of activity: C12 branched-chain amine greater than C12 straight-chain amine greater than C14 amine greater than C16 amine. The C12 branched amine also blocked the C-24(28)-dehydrogenase system in the conversion of sitosterol to fucosterol, the initial step in sitosterol dealkylation.

Amides↗