Heinz Bodies Formed in Erythrocytes by Fatty Acid Ozonides and Ozone

Ozonides of the methyl esters of oleic, linoleic, linolenic and arachidonic acids were found to produce Heinz body inclusions in human and mouse erythrocytes. No simple relationships between structure and activity were noted.

Concomitant with Heinz body formation, methemoglobin and loss of cellular thiols were observed. Methyl ozonides readily oxidized glutathione and 1 mole of oxidized glutathione was formed per mole of methyl oleate ozonide. Methyl ozonides catalyzed the formation of disulfide-linked interchain polymers between hemoglobin and ovalbumin. Heinz bodies were not produced with ozone in the absence of unsaturated lipids. Heinz bodies were observed in the blood of mice exposed to ozone (0.85 ppm) for 48 hours.

These observations suggest that fatty acid ozonides could serve as a toxic chemical species formed on ozone inhalation and could explain the divergent protective effects of lipid antioxidants and thiol generating systems in vivo.     

Prevention of Ozonide-Induced Heinz Bodies in Human Erythrocytes by Vitamin E

Methyl oleate ozonide, a proposed intermediary in ozone intoxication, produced Heinz body inclusions in human erythrocytes at concentrations of from 10^?4 to 2×10^?3 M. Daily oral supplementation with either 100 mg or 200 mg of d-α- tocopheryl acetate prevented Heinz body formation by methyl oleate ozonide.

These observations suggest that the protective effects of vitamin E against ozone-produced toxicants occurs in man as well as in animals.     

Carbon Monoxide and Physical Work Capacity

Submaximal and maximal exercise capacity were compared in 16 subjects while breathing air and a 225-ppm carbon monoxide-air mixture, the latter resulting in a carboxyhemoglobin level of 20%. Maximal oxygen uptake decreased approximately proportional to the reduction in arterial oxygen content (24%) and irrespective of the arterial oxygen tension. The Incremental increases of heart rate and ventilation with exercise were greater in the subjects when breathing CO than air but peak ventilation was reduced with CO. Except for the latter, these responses are similar to that found with hypobaric hypoxia.     

Utensil Swab Test

The effectiveness of the utensil swab test as part of the periodic inspection of food establishments was studied both as an indicator of cleanliness during the period between inspections and as an educational tool.

Two to seven samplings were taken at intervals of two to three months at 95 food establishments in rural areas in the northern part of Israel. A statistical analysis showed a high variation and almost no correlation between repeated samplings of the same establishment. Also, no trend to suggest an educational effect was found. We concluded that, though the swab test had proved to be useful in special projects, it had no value as part of the periodic sanitary inspections.     

Response of Cats to Inhaled Mixtures of SO2 and SO2-NaCl Aerosol in Air

Twenty healthy, adult male cats were lightly anesthetized with pentobarbital (Nembutal) sodium and tracheotomized; respiration was maintained by pump, Medical-grade breathing air, with or without sulfur dioxide alone or in combination with sodium chloride aerosol was delivered in predetermined exposure sequencer Pulmonary flow resistance and lung compliance were evaluated. Pollutants were also delivered via endotracheal catheter or face mask, or both. Approximately 20 ppm of SO₂ in air was required to evoke a significant change in pulmonary flow resistance in “reactors.” The majority of animals showed no response to this concentration of SO₂ either alone or with NaCl aerosol (10 mg/cu m). An increased frequency of significant changes in pulmonary flow resistance was suggested when pollutants were delivered via endotracheal catheter or face mask. All alterations in parameters of response were reversible shortly after exposure ceased.     

p-Amminobenzoic Acid as a Protective Agent in Ozone Toxicity

The ability of p-aminobenzoìc acid (PABA) to profeet against ozone toxicity was demonstrated in two experimental systems p-Aminobenzoic acid, and the related compounds p-aminohippuric acid and anthranïlïc acid, inhibited the ozone-induced loss of activity of the enzyme acetylcholinesterase in human erythrocytes exposed in vitro to 40 ppm of ozone. Protection in vivo was demonstrated In a study in which 15 rats injected intraperitoneally with 2 ml of 20 mM PABA prior to ozone exposure died in a mean of 426 minutes as compared to a mean of 261 minutes for 15 control animals similarly exposed to 15 ppm of ozone. These findings provide additional presumptive evidence of the importance of free radicals in the mechanism of ozone toxicity. They further suggest that PABA and related compounds warrant exploration as possible protective agents in susceptible populations exposed to significant photochemical air pollution.