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Background: Laser lesions may induce retinal damage that is larger than expected from the size of the coagulated area. This study was designed to follow the development of laser-induced reduction in retinal function and to correlate it with structural changes., Methods: Pigmented rabbits were treated in one eye with 225 argon laser lesions. The ERG responses were recorded at different times after treatment. The effect of the laser treatment upon the functional integrity of the retina was assessed from the ERG responses. Structural damage was examined by light microscopy., Results: Shortly (1-2 h) after laser treatment, the ERG responses were reduced by about 50%. ERG deficit continued to develop and reached a maximal level about 24 h after treatment. Thereafter, slow recovery was observed but permanent deficit, relative to the initial laser effect, was seen even 30 days after treatment. Histological observations indicated extensive serous retinal detachment between laser lesions that developed within 24 h after treatment. At 30 days post-treatment, lesioned areas were completely destroyed and heavily pigmented. The retina between lesions was attached to the pigment epithelium but exhibited different degrees of structural damage., Conclusions: The immediate laser damage is confined to the coagulated areas while secondary functional damage develops within 24 h and probably reflects serous retinal detachment between lesions. The serous retinal detachment completely resolves with time but may induce permanent structural abnormalities in non-coagulated retinal areas that is reflected in a functional deficit larger than the initial laser effect.

The involvement of the L-arginine-nitric oxide (NO) pathway in the pathogenesis of hyperoxia-induced seizures was studied by using agents controlling NO levels. We selected two inhibitors of nitric oxide synthase, the systemic inhibitor Nomega-nitro-L-arginine methyl ester (L-NAME) and the novel cerebral-specific inhibitor 7-nitroindazole, and two generators of NO, the NO donor S-nitroso-N-acetylpenicillamine and the physiological precursor L-arginine. Rats with chronic cortical electrodes were injected intraperitoneally with different doses of one of the agents or their vehicles before exposure to 0.5 MPa O2 and O2 with 5% CO2 at an absolute pressure of 0.5 MPa. The duration of the latent period until the onset of electrical discharges in the electroencephalogram was used as an index of central nervous system O2 toxicity. The two nitric oxide synthase inhibitors L-NAME and 7-nitroindazole significantly prolonged the latent period to the onset of seizures on exposure to both hyperbaric O2 and to the hypercapnic-hyperoxic mixture. Pretreatment with the NO donor S-nitroso-N-acetylpenicillamine significantly shortened the latent period, whereas L-arginine, the physiological precursor of NO, significantly prolonged the latent period to onset of seizures. Our results suggest that the L-arginine-NO pathway is involved in the pathophysiology of hyperoxia-induced seizures via various regulating mechanisms.

We investigated the secretion of TNF-alpha by monocytes and macrophages derived from the peripheral blood, spleen, and lungs after a single exposure to a therapeutic profile of hyperbaric oxygen (HBO). Rats were exposed for 90 min to either 100% oxygen at 0.28 MPa (2.8 atmospheres absolute) or air. Immediately after exposure, mononuclear cells were isolated from blood, spleen, and lungs and cultured for 18 h. The secretion of TNF-alpha from the cultured monocytes/macrophages was determined with and without stimulation with lipopolysaccharide (LPS). Exposure to hyperbaric oxygen induced a significant increase in the spontaneous ex vivo secretion of TNF-alpha (without LPS) by mononuclear cells from the blood, spleen, and lung (P < 0.05 from air controls). Stimulation with LPS after exposure to HBO induced a significant increase in TNF-alpha secretion by lung and spleen macrophages compared with air controls (P < 0.05). However, absolute TNF-alpha levels were not significantly higher than those achieved 'spontaneously' in macrophages exposed to HBO without LPS. Stimulation with LPS induced a marked increase in secretion of TNF-alpha from blood monocytes after exposure to air, but not after exposure to HBO. These results provide evidence in support of a role played by TNF-alpha in mediating HBO effects on different tissues and their immune responses.

We tested the effect of caffeine, on hyperoxia-induced seizures. Thirty-seven rats with chronic cortical electrodes were injected i.p. with caffeine (1.25, 2.5, and 10 mg/kg) or vehicle before exposure to 0.5 MPa oxygen and 17 rats to oxygen with 5% CO2 at 0.5 MPa. EEG monitoring and spectral analysis of EEG activity were carried out. Caffeine significantly prolonged the latent period to the onset of seizures (P < 0.05 in ANOVA), in a dose-related manner. Our results suggest that caffeine may be used in low doses for protection against hyperoxia-induced seizures.

The toxicity of hyperbaric oxygen in the central nervous system is expressed by generalized tonic-clonic seizures. In the search for drugs effective against these seizures, we tested flumazenil, a benzodiazepine antagonist known to have antiepileptic properties. Forty rats with chronic cortical electrodes were injected i.p. with one of three doses of flumazenil (0.2, 2, and 20 mg/kg) or their vehicle, before exposure to 0.5 MPa oxygen. EEG and the spectral analysis of background EEG activity were monitored. The duration of the latent period until the appearance of electrical discharges in the EEG was used as an index of oxygen toxicity. Significant prolongation of the latent period preceding oxygen seizures was noted in the group of rats treated with 0.2 mg/kg flumazenil (p < 0.05 in Tukey test). However, when the dose of flumazenil was increased, the duration of the latent period returned to control values. These diverse effects of flumazenil on the benzodiazepine receptor may account for its complex effects on hyperoxic-induced seizures.

Decreased oxygen delivery and cellular hypoxia are major factors in the pathophysiology of shock. We studied the effects of 100% O2 at 0.1 and 0.3 MPa (1 and 3 atm abs) in severe hemorrhagic shock in awake, unrestrained rats. Shock was induced by withdrawing 50% of the total blood volume within 120 min. Blood pressure, heart rate, and the electroencephalogram (EEG) were recorded during the first 6 h of the protocol. The animals were observed for 7 days. The shock protocol resulted in 60 and 90% mortality after 1 day and at the end of 7 days, respectively. A single 90-min exposure to O2 at 0.1 and 0.3 MPa, which was started 30 min after bleeding, maintained mean arterial blood pressure at significantly higher values compared to untreated controls throughout the exposure period (P < 0.05). Oxygen therapy at both doses also improved the long-term survival rate and survival time significantly (P < 0.01). No clinical or EEG sign of CNS O2 toxicity was detected in O2-treated animals. Our results indicate that O2 given alone after severe bleeding exerts a beneficial effect on the long-term outcome of hemorrhagic shock in awake, unrestrained rats.

We studied the effect of a single exposure to a therapeutic profile of hyperbaric oxygen on blood mononuclear cell subset. Twenty healthy volunteers were exposed to 0.28 MPa for 90 min. Thirteen breathed pure oxygen and seven were control subjects exposed to compressed air at the same pressure. Venous blood samples were drawn before HBO exposure, immediately on exit from the chamber, and 24 h later. Immediately after the exposure, a significant increase was observed in the percentage and absolute number of CD8 (suppressor/cytotoxic) T cells, with a concomitant decrease in the CD4 (helper/inducer) T cells. These changes resulted in a decreased CD4:CD8 ratio. A rise was also observed in the number of HLA-DR antigen-bearing cells, with a transient increase in monocytes. There was no change in the total count and percentage of T cells (CD3), B cells, and NK cells. Twenty-four hours after HBO exposure there was a partial reversal of the decrease in the mean CD4:CD8 ratio, but it was still significantly lower than preexposure values. The fast reversibility of the change in the CD4:CD8 ratio suggests specific HBO-induced shifts and sequestration of T-cell subpopulations.

The toxicity of hyperbaric oxygen in the central nervous system is expressed by generalized tonic-clonic seizures. In the search for drugs effective against these seizures, we tested flunarizine, a calcium antagonist known to have antiepileptic properties and only minimal cardiovascular side effects. 49 rats with chronic cortical electrodes were injected i.p. with six different doses of flunarizine (10-300 mg/kg) or vehicle, before exposure to 0.5 MPa oxygen. Two doses of flunarizine and vehicle were given to rats exposed to oxygen with 5% CO2 at an absolute pressure of 0.5 MPa. EEG and spectral analysis of background EEG activity were monitored. The duration of the latent period before the appearance of electrical discharges in the EEG was used as an index of oxygen toxicity. There was no statistical difference between the duration of the latent periods for the seven groups treated by flunarizine or by vehicle on exposure to 0.5 MPa pure oxygen (P = 0.9 in ANOVA), but on exposure to oxygen with CO2, there was significant prolongation of the latent periods in comparison with vehicle (P < 0.001). Our results suggest that on exposure to hyperbaric oxygen, the antiepileptic effect of flunarizine might be masked, probably by its cerebral antivasoconstrictive effect.

The toxicity of hyperbaric oxygen in the central nervous system is expressed by clinical and electroencephalographic (EEG) manifestations resembling those of generalized tonic-clonic seizures. In the search for drugs effective against these seizures, we tested vigabatrin, an irreversible inhibitor of GABA (gamma-aminobutyric acid) transaminase. Five different doses of vigabatrin (ranging from 50 to 500 mg/kg) or vehicle were injected i.p. in rats implanted with cortical electrodes, 4 h prior to exposure to 5 ATA (0.5 MPa) oxygen. EEG and spectral analysis of the background EEG activity were monitored for the different dosages of the drug. The duration of the latent period before the appearance of electrical discharges in the EEG was used as an index of oxygen toxicity. The protective effect of vigabatrin was dose-related, and complete protection against hyperoxic-induced discharges was at 180 mg/kg. The protective effect lasted 24 h and decreased gradually disappearing completely on the third day. An increase in the low frequency bands of the EEG and a decrease in the faster activity were correlated with the vigabatrin dosage injected. Our results suggest that vigabatrin has the potential of being a useful drug in the treatment and prevention of oxygen-induced seizures during hyperbaric oxygen therapy.

Scopolamine (Hyoscine), an anticholinergic compound is widely used for the prophylaxis and treatment of motion sickness and might be used with oxygen diving and hyperbaric oxygen therapy. We therefore decided to test the interaction of scopolamine with oxygen at high pressure. Thirty-six rats implanted with cortical EEG electrodes were injected subcutaneously with two doses of scopolamine (0.02 or 0.2 mg.kg-1), or the vehicle (saline), 30 min before exposure to 5 atm abs (0.5 MPa) oxygen. Electroencephalogram and heart rate were monitored continuously. Spectral analysis of the EEG was carried out, and the duration of the latent period before convulsions was determined. No significant difference was found in the duration of the latent period between the control rats receiving vehicle (saline) and rats injected with scopolamine (n = 12 for each group). Changes in background EEG activity and maximal dilation of the pupil were detected at both scopolamine doses. Heart rate significantly decreased at 0.02 mg.kg-1 and increased at the dose of 0.2 mg.kg-1 scopolamine. Our findings indicate that the duration of the latent period preceding hyperoxic seizures is not altered by scopolamine in rats; however, other side effects of the drug regarding visual and cardiovascular symptoms should be considered when scopolamine is used in combination with hyperbaric oxygen.

The main manifestations of CNS oxygen toxicity are generalized tonic-clonic seizures. We tested the protective effect of 2 antiepileptic drugs, carbamazepine and ethosuximide, which are commonly used for the treatment of generalized seizures, on hyperbaric oxygen-induced convulsions. Rats implanted with chronic cortical electrodes for continuous EEG monitoring were injected i.p. with either carbamazepine (5 doses in the range of 1.5-50 mg/kg), ethosuximide (400 mg/kg), or their vehicles (40% propylene glycol and saline, respectively). The rats were exposed to 5 ATA (0.5 MPa) oxygen. The duration of the latency until the appearance of electrical discharges in the EEG was used as an index of toxicity. Ethosuximide did not protect against hyperoxic seizures. In contrast, rats pretreated (30 min) with carbamazepine exhibited a dose-related protective effect against hyperoxically induced seizures. The results of our study suggest that carbamazepine should be considered for prevention of oxygen-induced seizures during hyperbaric oxygen therapy.

Decreased oxygen delivery and cellular hypoxia are important factors in the pathophysiology of hemorrhagic shock. We studied the effects of 100% oxygen at 1 and 3 ATA (atmosphere absolute) in a severe model of hemorrhagic shock induced by bleeding 50% of the total blood volume in rats. Post-treatment with 100% oxygen at 1 and 3 ATA maintained mean arterial blood pressure (MABP) in hemorrhaged rats at significantly higher values compared to untreated controls (P less than 0.01 at 1 and 3 ATA). Treatment with oxygen attenuated the increase in plasma activities of the lysosomal hydrolase cathepsin D (P less than 0.05 at 1 ATA; P less than 0.01 at 3 ATA). Oxygen at 3 ATA also attenuated the plasma accumulation of free amino-nitrogen compounds (P less than 0.05). Furthermore, hyperoxia prevented the final increase in hematocrit (P less than 0.05 at 1 ATA; P less than 0.01 at 3 ATA). Hemorrhaged rats treated with oxygen also exhibited a significantly longer survival time (P less than 0.01 at both doses), and higher survival rates (50% at 1 ATA and 100% at 3 ATA; P less than 0.05 and P less than 0.01, respectively) than untreated shock rats. No significant effect on any of the above mentioned variables was found in hemorrhaged rats treated with 7% oxygen at 3 ATA (oxygen pressure 0.2 ATA), indicating that all salutary effects can be attributed to oxygen and not to the increased ambient pressure per se. Our results indicate that 100% oxygen in normobaric and hyperbaric conditions exerts important beneficial effects in hemorrhagic shock and may be a useful drug for the treatment of this condition.

Animal models are widely used for the study of CNS oxygen toxicity, but confusion still exists regarding the proper statistical approach to the analysis of the data. This paper is based on data collected from unanesthetized, free-moving rats with chronically implanted cortical electrodes for continuous EEG monitoring, exposed to 5 or 6 ATA oxygen. The index measured for CNS oxygen toxicity is the duration of the latent period preceding the appearance of well-defined electrical discharges in the EEG. At both oxygen pressures studied, the duration of the latent period is not distributed normally, and variability within the groups is not homogeneous. Transformations of the latent period data were found to enhance normality, and the speed of appearance of the discharges in the EEG, which is the reciprocal of the time, seems to be a simple, useful index for CNS oxygen toxicity in rats. Two experimental designs were compared: repeated measurements vs. single exposure. No advantage was demonstrated for the use of each rat as its own control as against the comparison between data from groups of rats. Rats can be used more than once in such research, but not more than once in a single study where individual observations are assumed to be independent, since there is some positive correlation between the first and second exposures to hyperbaric oxygen in individual rats; however, the level of sensitivity of the groups is not significantly different.

The effect of ambient illumination on sensitivity to CNS oxygen toxicity was studied in awake male rats. Continuous recording of the EEG was obtained with chronically implanted cortical electrodes. The appearance of the electrical discharges in the EEG was used as an end point for CNS oxygen toxicity. It was found that sensitivity to CNS oxygen toxicity was inversely related to the level of ambient light illumination. The latent period for appearance of the electrical discharges was significantly shorter in darkness than in light over most of the oxygen pressures. Rats with severely depressed retinal function were as sensitive to oxygen at high pressure as normal rats in darkness, demonstrating the importance of visual input in the modulation of sensitivity to CNS oxygen toxicity.

The effect of sodium phenytoin on hyperbaric oxygen (HBO) induced central nervous system (CNS) toxicity was studied in rats. The latency of the epileptic electroencephalographic discharges was measured in 20 phenytoin-treated rats exposed to 6 ATA of 100% oxygen, and compared with that of 20 saline-injected rats exposed to the same pressure of pure oxygen. No statistically significant difference was found in the latency between the two groups. In addition, phenytoin failed to suppress or to modify the clinical seizures. Sodium phenytoin failed to suppress or to modify the clinical seizures. Sodium phenytoin blood levels were determined in the rats, and were found to be within the therapeutic range. We conclude that sodium phenytoin is ineffective in suppressing HBO induced CNS oxygen toxicity.

We studied the effects of hyperbaric oxygen in a severe model of circulatory shock induced by occlusion and reperfusion of major splanchnic arteries (splanchnic artery occlusion (SAO) shock). Pentobarbital-anesthetized rats subjected to total occlusion of the superior mesenteric and the celiac arteries for 40 min developed a severe shock state, resulting in a uniformly fatal outcome after release of the occlusion. Exposure to hyperbaric oxygen at 2 ATA (atmosphere absolute) (1 ATA = 0.1 MPa) was initiated immediately after reperfusion. SAO shock rats exposed to hyperbaric oxygen maintained mean arterial blood pressure at significantly higher values throughout the postreperfusion period compared with untreated SAO shock rats (p less than 0.01), with final mean arterial blood pressures of 88 +/- 9 and 51 +/- 4 mmHg, respectively. Treatment with hyperbaric oxygen attenuated the increase in plasma activities of the lysosomal hydrolase cathepsin D (p less than 0.05), and diminished the increase of hematocrit (p less than 0.01 from untreated shock rats). Splanchnic occlusion shock rats treated with hyperbaric oxygen also exhibited a significantly higher survival rate than the untreated shock group (77 vs. 0%, respectively; p less than 0.01). Our results suggest that the beneficial effects of exposure to hyperbaric oxygen immediately after reperfusion of the splanchnic region outweigh its possible deleterious effect.

A decrease in brain GABA concentration has been implied as the cause of convulsions induced by hyperbaric oxygen (HOP). We therefore examined the influence of sodium valproate, an anticonvulsant and GABA transaminase inhibitor on HOP-induced convulsions in rats. The mean latency of occurrence of the first electrical discharge in the ECoG and the appearance of the first clinical seizure in awake chronically implanted rats was unchanged by administration of sodium valproate prior to HOP exposure. We conclude that either the sodium valproate inhibition of GABA removal is insufficient to compensate for HOP inhibition of its production, or else that GABA concentration changes are not causally related to HOP-induced seizures.

Central nervous system oxygen toxicity in hyperbaric oxygen-inert gas mixtures was studied by exposing male rats to various gas mixtures having the same oxygen partial pressure and varying pressures of inert gases. The duration of the latent period until the appearance of electrical discharges in the electroencephalogram was used as the criterion for the sensitivity to CNS oxygen toxicity. Two hundred and twenty rats with chronically implanted cortical electrodes were subjected to a single exposure to 1 of 11 different gas mixtures at ambient pressures ranging from 5 to 10 ATA. All gas mixtures had a constant oxygen partial pressure of 5 ATA and varying pressures of inert gas (helium or nitrogen). The duration of the latent period was found to be significantly different in the 11 experimental groups (P less than 0.001). While increasing the inert gas pressure, the latency gradually shortened, reaching the lowest values in mixtures of 5 ATA oxygen and 3 ATA of either inert gas. On further increase in the inert gas pressure up to a total ambient pressure of 10 ATA, the latency returned to control (pure 5 ATA oxygen) values. No difference was found between nitrogen and helium of equal partial pressures. These findings suggest that the risk of CNS oxygen toxicity in gas mixtures is not determined solely by the PO2, and even a relatively low pressure of inert gas can contribute to the imminence of hyperbaric-oxygen-induced seizures.