Search

Expired ethane is regarded as a noninvasive indicator of lipid peroxidation. As a model of oxidative stress we have investigated in male Wistar rats (body wt. 309 ± 15 g) the effects of various levels of elevated inspiratory oxygen concentrations on the expiration rate of ethane. After 4 days under 21 vol% O2 (basic condition) the rats were exposed for 6 or 5 days to 40, 60 or 80 vol% O2 over 8 or 23 h/day. The variously O2-enriched air was conducted through the cages and expired ethane adsorbed onto charcoal was thermo-desorbed and measured by gas chromatography. Basic ethane expiration was 3.1 ± 0.8 pmol/100 g body wt. per min. At 40 vol% O2 over 8 or 23 h/day no increase or a maximum average 47% increase (P < 0.01) in ethane expiration occurred on day 4; 60 vol% O2 over 8 or 23 h/day led to a corresponding increase of 56 or 87% (P < 0.05 or P < 0.01) on day 3; 80 vol% O2 over 8 or 23 h/day led to a corresponding increase of 81 or 66% (P < 0.01) on days 3 or 2. Our results indicate that with up to 60 vol% O2 a temporary increase in lipid peroxidation occurs in a dose dependent manner. However, at 80 vol% O2 no further increase in the maximum ethane expiration occurred. The latter finding and the finding of only transient increase in ethane expiration in probably due to antioxidative counteraction.

The bioactivation and binding of 14C-labelled 1,1,2,2-tetrachloroethane (TCE) in the tissues of C57B1 mice were studied. As shown by autoradiography with heated and organic solvent-extracted tissue sections of i.v. injected mice, a high and selective localization of bound metabolites occurred in the nasal olfactory mucosa, preferentially in the Bowman's glands. High levels of bound metabolites were also present in epithelia of the trachea, bronchi and bronchioli and in the squamous epithelia of the oral cavity, tongue and esophagus. An epithelial binding was observed in tissue slices incubated with 14C-TCE. Incubation of 14C-TCE with homogenates of the olfactory mucosa and liver showed that the olfactory mucosa had a higher ability to activate 14C-TCE into products that become irreversibly bound to protein. Addition of metyrapone, glutathione or sodium dithionite to the incubations decreased the level of irreversible binding, suggesting that the activation of TCE to reactive products is mediated via an oxidative cytochrome P-450 dependent process in the olfactory mucosa.

Mice, rats and guinea pigs were exposed to the smoke produced by ignition of a zinc oxide/hexachloroethane pyrotechnic composition, 1 h/day, 5 days/week, at three different dose levels, together with controls. The animals received 100 exposures except for the high dose guinea pigs, which underwent 15 exposures, because of high death rate during the first few days of exposure. The test material had very little effect on weight gain, but there was a high rate of early deaths in the top dose of mice. A variety of incidental findings was seen in both decedents and survivors, but organ specific toxicity was, with one exception, confined to the respiratory tract. The most important of these findings was a statistically significant increase in the frequency of alveologenic carcinoma in the high dose group mice (p less than 0.01) and a statistically significant trend in the prevalence of the same tumour over all dose groups and the controls. A variety of inflammatory changes was seen in the lungs of all species and some appeared to be treatment-related. Fatty change in the mouse liver was more common in the middle and high dose groups than the controls. The aetiology of the tumour incidence is discussed and it is pointed out that hexachloroethane and zinc, as well as carbon tetrachloride, which may be present in the smoke, may be animal carcinogens in appropriate circumstances. Carbon tetrachloride is a known human carcinogen.

Isolated rat hepatocytes (1 X 10(7) cells/ml) were aerobically incubated in Eagle's Minimum Essential Medium which contained 2.0% albumin. As potential parameters of lipid peroxidation ethane and n-pentane formed were measured in samples obtained from the gas phase above the incubation mixture. 15-30 nmol ethane or n-pentane were produced by 10(7) hepatocytes within 90 min. Carbon tetrachloride (CCl4) or ADP-complexed ferrous ions stimulated ethane and n-pentane formation considerably, depending on the concentrations of the compounds. With CCl4 10(7) cells formed max 180 nmol ethane and 140 nmol n-pentane within 90 min incubation, whereas with Fe(II) max 130 nmol ethane and 220 nmol n-pentane could be detected. When n-pentane was added to the gas phase above the incubation mixture containing either medium or medium plus hepatocytes its amount decreased by 30% within the first 5 min of incubation. However, afterwards only minor amounts of n-pentane disappeared, even in the presence of hepatocytes. This indicates that n-pentane equilibrates with the cells suspension under the conditions used. Cell viability, as determined by the release of lactate dehydrogenase into the medium and by the uptake of trypan blue by the cells, and the recovery of the cells decreased only in presence of relatively high concentrations of CCl4, or Fe(II) respectively. However, a maximal effect on ethane and n-pentane formation was reached already with lower concentration.

Ethane exhalation was increased in male Sprague-Dawley rats following a single intraperitoneal (IP) injection of Aroclor 1254 (500 mg/kg). In the first 2 weeks following Aroclor 1254 treatment, the increase in ethane exhalation was due to an inhibition of metabolism of endogenous ethane rather than to an increase in ethane production. In weeks 3 and 4 following Aroclor 1254 administration, metabolic clearance of ethane returned to and exceeded control levels, while ethane production increased to approximately twice the control rates (day 30). The HPLC determination of in situ hepatic malondialdehyde levels revealed a 2-fold increase in malondialdehyde content on day 30 following the Aroclor 1254 injection. Further, parallel increases in in situ malondialdehyde levels and ethane production rates were also found 30 days following a single IP injection of 3,3',4,4'-tetrachlorobiphenyl, 2,3,4,4',5-pentachlorobiphenyl and 2,2',4,4',5,5'-hexachlorobiphenyl (300 mumol/kg). These effects were not reflected in increased diene conjugation. Redox state of the liver was largely unaffected, as evidenced by the relative concentrations of reduced and oxidized NADPH. However, minor changes in reduced and oxidized glutathione were noted.

The investigation was aimed at providing insight into the side chain metabolism of furazolidone in mice. The agents used in the experiments were furazolidone, 3-amino-2-oxazolidinone, oxazolidinone, nitrofurantoin, 3-aminohydantoin and hydantoin, administered intraperitoneally at five equimolar doses ranging from 0.178 to 0.888 mmol/kg. The parameters investigated included ethane and ethylene expiration, formation of malondialdehyde and total glutathione content in the liver. Ethylene expiration was found to be strongly enhanced by aminooxazolidinone and slightly increased by furazolidone. Ethane expiration was increased after aminooxazolidinone administration. Malondialdehyde formation was not affected by any of the agents used. Total glutathione was decreased by furazolidone and nitrofurantoin. The above findings indicate that, in vivo, the azomethine linkage of the side chain of furazolidone hydrolyses to form 3-amino-2-oxazolidinone, subsequently cleaved to ethylene.

The antidote efficacy of liquid paraffin in oral solvent intoxications was investigated in dogs. Groups of 3 to 4 animals each received 1 ml/kg of methylchloroform, 0.3 ml/kg of benzene, or 3 ml/kg of a synthetic gasoline mixture (hexane/heptane/octane, ratio 30:50:20) by gastric intubation. After a prolonged interval the same animals were given 5 ml/kg of liquid paraffin by a separate tube in addition to one of the above solvents. The solvent concentrations in the blood were determined by gas chromatography at appropriate intervals. Administration of liquid paraffin in conjunction with methylchloroform does not significantly affect the absorption process. In the case of benzene and the synthetic gasoline mixture, however, liquid paraffin produces a distinct reduction in the rate of absorption. It is thus concluded that in the therapy of oral solvent-intoxication, liquid paraffin exhibits favorable antidote properties which may however differ from one solvent to the other. In clinically relevant situations involving coadministration of purgatives, the antidote effect must be assessed as being of a higher order than in our animal experiments in which purgatives had to be omitted for external reasons.

The exhalation of ethane is widely used as an indicator of in vivo lipid peroxidation. To test the hypothesis that lipid peroxidative events are involved in the toxicity of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD), we administered a lethal dose of TCDD (60 μg/kg), IP to male Sprague Dawley rats (160–180 g) and measured by gas chromatography the exhalation of ethane into the atmosphere of a closed all-glass exposure chamber. TCDD-treated rats exhaled only slightly more ethane than control rats at a single time point 7 days following TCDD administration. Since the exhalation of ethane is the net result of the endogenous production of the gas and its metabolic degradation, the latter was quantified by measuring the clearance of exogenous ethane (initial concentration = 100 ppm) introduced to the atmosphere of the exposure chamber. The clearance of ethane in TCDD-treated rats was markedly decreased, reaching a minimum 7 days following TCDD treatment. Apparently, the slight increase in exhaled ethane was due to an inhibition of ethane metabolism caused by TCDD. However, rats obviously intoxicated and having lost considerable body weight might be impaired in their ability to transport ethane. To bypass this problem we injected ethane (0.2 ml) directly into the rats IP. Here also the metabolic clearance in TCDD-treated rats was diminished. In a further experiment, rats treated with dithiocarb at a dose where ethane metabolism was totally inhibited exhaled more ethane than did TCDD-treated rats. It is therefore concluded that the slight increase in ethane exhalation following a lethal dose of TCDD is due to a partial inhibition of ethane metabolism and that there is no net increase in ethane production due to lipid peroxidation. Indeed when TCDD-treated rats were administered Fe++, a well-known initiator of lipid peroxidation, they were less competent to carry out lipid peroxidation than rats treated with Fe++alone.

Rats were exposed to white smoke generated from mixtures of titanium dioxide-hexachloroethane (TiO2-HC) and zinc-hexachloroethane (Zn-HC), respectively, in an inhalation chamber operated in the static mode. The dose was varied by varying the amount of smoke mixture and/or the exposure time. The acute inhalation toxicity of TiO2-HC smoke was much lower than the Zn-HC smoke. Thus, the animals survived exposure to TiO2-HC smoke, even at relatively high smoke concentrations. This smoke was irritating to the animals and minor, acute inflammatory changes were seen in lung tissue. In contrast, Zn-HC smoke was very toxic and caused lethal injuries to the experimental animals, even at relatively low concentrations. Pulmonary injuries were extensive and death was due to blood congestion with pulmonary oedema. Since the TiO2-HC and Zn-HC mixtures form TiCl4 and ZnCl2, respectively, a separate study was performed in which rats were exposed to TiCl4 gas or ZnCl2 aerosol. No animals died from exposure to TiCl4 at concentrations between 370 and 2900 mg/m3 for 10 min. The LC50 of ZnCl2 was found to be around 2000 mg/m3 during a 10-min exposure period. The difference between the two types of smoke is explained by the difference in toxicity between TiCl4 and ZnCl2.

About 250 mumol/kg tetrachloromethane is metabolized by male Sprague-Dawley rats receiving a dose of 500 mumol/kg by inhalation. Determination of enzyme activities and histopathological assessment show that various parameters reflecting cellular injury do not differ for the same amount of tetrachloromethane metabolized under different oxygen partial pressures. On the other hand, the amounts of ethane and pentane exhaled under hypoxic conditions are greater than under normoxia. The previously reported differences in the toxicity of tetrachloromethane upon exposure under normoxia or hypoxia seem to be mainly due to the different amounts of tetrachloromethane metabolized under both conditions.

The use of exhalation of ethane and n-pentane in experimental animals as parameters of lipid peroxidation led to an examination of pharmacokinetics of both compounds in rats. When rats were exposed, in a closed desiccator jar chamber, to a wide range of ethane concentrations, linear elimination pharmacokinetics were observed. n-Pentane, when concentrations higher than 100 ppm were applied, displayed saturation kinetics. These were formally explained by action of two competing metabolizing pathways or enzymes. Application of preexisting models could describe exhalation of both ethane and n-pentane by untreated control rats. Stimulation of lipid peroxidation by ferrous ions or by carbon tetrachloride resulted in dissimilar quantitative behaviours of ethane and n-pentane. Ethane production rates were enhanced after application of both compounds. Because of relatively slow metabolic eliminations this led to markedly elevated concentrations of ethane in the gas phase of the system. Pentane production rates were simultaneously enhanced. However, difficulties in interpretation arise because of rapid metabolic elimination of n-pentane. Compounds that diminish pentane metabolism are shown to evoke higher pentane concentrations in the system than compounds which only enhance the pentane production rate. Determinations of ethane exhalation should provide a more favourable parameter of lipid peroxidation than exhalation of pentane.