Acclimation to decompression sickness in rats.

Protection against decompression sickness (DCS) by acclimation to hyperbaric decompression has been hypothesized but never proven. We exposed rats to acclimation dives followed by a stressful "test" dive to determine whether acclimation occurred. Experiments were divided into two phases. Phase 1 rats were exposed to daily acclimation dives of hyperbaric air for 30 min followed by rapid decompression on one of the following regimens: 70 ft of seawater (fsw) for 9 days (L70), 70 fsw for 4 days (S70), 40 fsw for 9 days (L40), 40 fsw for 4 days (S40), or unpressurized sham exposure for 9 days (Control). On the day following the last exposure, all were subjected to a "test" dive (175 fsw, 60 min, rapid decompression). Both L70 and S70 rats had significantly lower incidences of DCS than Control rats (36% and 41% vs. 62%, respectively). DCS incidences for the other regimens were lower than in Control rats but without statistical significance. Phase 2 used the most protective regimen from phase 1 (L70); rats were exposed to L70 or a similar regimen with a less stressful staged decompression. Another group was exposed to a single acclimation dive (70 fsw/30 min) on the day before the test dive. We observed a nonsignificant trend for the rapidly decompressed L70 dives to be more protective than staged decompression dives (44% vs. 51% DCS incidence). The single acclimation dive regimen did not provide protection. We conclude that protection against DCS can be attained with acclimating exposures that do not themselves cause DCS. The deeper acclimation dive regimens (70 fsw) provided the most protection.

Using animal data to improve prediction of human decompression risk following air-saturation dives.

To plan for any future rescue of personnel in a disabled and pressurized submarine, the US Navy needs a method for predicting risk of decompression sickness under possible scenarios for crew recovery. Such scenarios include direct ascent from compressed air exposures with risks too high for ethical human experiments. Animal data, however, with their extensive range of exposure pressures and incidence of decompression sickness, could improve prediction of high-risk human exposures. Hill equation dose-response models were fit, by using maximum likelihood, to 898 air-saturation, direct-ascent dives from humans, pigs, and rats, both individually and combined. Combining the species allowed estimation of one, more precise Hill equation exponent (steepness parameter), thus increasing the precision associated with human risk predictions. These predictions agreed more closely with the observed data at 2 ATA, compared with a current, more general, US Navy model, although the confidence limits of both models overlapped those of the data. However, the greatest benefit of adding animal data was observed after removal of the highest risk human exposures, requiring the models to extrapolate.

Mixed-gas model for predicting decompression sickness in rats.

A mixed-gas model for rats was developed to further explore the role of different gases in decompression and to provide a global model for possible future evaluation of its usefulness for human prediction. A Hill-equation dose-response model was fitted to over 5,000 rat dives by using the technique of maximum likelihood. These dives used various mixtures of He, N(2), Ar, and O(2) and had times at depth up to 2 h and varied decompression profiles. Results supported past findings, including 1) differences among the gases in decompression risk (He < N(2) < Ar) and exchange rate (He > Ar approximately N(2)), 2) significant decompression risk of O(2), and 3) increased risk of decompression sickness with heavier animals. New findings included asymmetrical gas exchange with gas washout often unexpectedly faster than uptake. Model success was demonstrated by the relatively small errors (and their random scatter) between model predictions and actual incidences. This mixed-gas model for prediction of decompression sickness in rats is the first such model for any animal species that covers such a broad range of gas mixtures and dive profiles.

Evaluation of oxygen and pressure in treatment of decompression sickness in guinea pigs.

These experiments examined whether increasing the partial pressure of oxygen (PO2), hydrostatic pressure, or both were responsible for the improvement in effectiveness of recompression treatment previously observed in guinea pigs with increasing depths of air. Unanesthetized male guinea pigs (600-700 g) were subjected to 8.6 atm abs (871 kPa) air dives for 60 min and then decompressed at 1.82 atm (184 kPa)/min to the surface. Subsequently, animals usually displayed hypotension, cardiac arrhythmia, and tachypnea, indicative of a fatal bout (> 95% death rate) of decompression sickness (DCS). Animals that developed DCS were treated by recompressing to depths ranging from 2.5 to 11.6 atm abs (253-1175 kPa), with 14, 28, 42, or 100% O2/balance N2. This design produced PO2's at treatment depth ranging from 0.4 to 3.6 atm abs (41-365 kPa). Upon recompression, recovery of blood pressure, heart rate, and breathing rate generally occurred. The area under the breathing rate vs. time curve was used to examine the effectiveness of treatment over a period of 60 min. A dramatic improvement in recovery over time was observed with increasing recompression depth for all gas mixtures. Analysis indicated that the positive response to depth was related to increasing hydrostatic pressure, increasing PO2 had no statistically significant beneficial effect.

Decompression comparison of helium and hydrogen in rats.

The hypothesis that there are differences in decompression risk between He and H2 was examined in 1,607 unanesthetized male albino rats subjected to dives on 2% O2-balance He or 2% O2-balance H2 (depths < or = 50 ATA, bottom times < or = 60 min). The animals were decompressed to 10.8 ATA with profiles varying from rapid to slow, with up to four decompression stops of up to 60 min each. Maximum likelihood analysis was used to estimate the relative decompression risk on a per unit pressure basis (termed "potency") and the rate of gas uptake and elimination, both factors affecting the decompression sickness risk, from a specific dive profile. H2 potency for causing decompression sickness was found to be up to 35% greater than that for He. Uptake rates were unresolvable between the two gases with the time constant (TC) estimated at approximately 2-3 min, leading to saturation in both cases in < 15 min. Washout of both gases was significantly slower than uptake, with He washout (TC approximately 1.5-3 h) substantially slower than H2 washout (TC approximately 0.5 h). It is unknown whether the decompression advantage of the faster washout of H2 or the disadvantage of its increased potency, observed in the rat, would be important for human diving.

Chemical safety of U.S. Navy Fleet soda lime.

Contamination was suspected of U.S. Navy Fleet soda lime (High Performance Sodasorb) when an ammonia-like odor was reported during its use in August 1992. This material contained indicator dye and was used for carbon dioxide absorption during diving. This incident had a major impact on the U.S Navy diving program when the Navy temporarily banned use of Sodasorb and authorized Sofnolime as an interim replacement. The Naval Medical Research Institute was assigned to investigate. Testing involved sampling from the headspace (gas space) inside closed buckets and from an apparatus simulating conditions during operational diving. Volatile organic compounds were analyzed by gas chromatography and mass spectrometry; ammonia and amines were measured by infrared spectroscopy. Significant amounts of ammonia (up to 30 ppm), ethyl and diethyl amines (up to several ppm), and various aliphatic hydrocarbons (up to 60 ppm) were detected during testing of both Sodasorb and Sofnolime. Contaminants were slowly removed by gas flow and did not return. The source(s) of the ammonia and amines are unknown, although they may result from the breakdown of the indicator dye. Hydrocarbon contamination seems to result from the materials of which the bucket is constructed. Unfortunately, evaluation of potential hazards associated with this contamination is difficult, due in part to the large number of variables of operational use and the absence of appropriate exposure limits. Based on these findings, the U.S. Navy has begun to phase in, for all diving, non-indicating soda lime that will be required to meet defined contaminant limits.

Intravascular bubble composition in guinea pigs: a possible explanation for differences in decompression risk among different gases.

Differences in risk of decompression sickness (DCS) that have been observed among inert gases may reflect differences in gas solubility or diffusivity or both. A higher risk gas might generate a larger volume of evolved gas during decompression, thereby increasing the probability of DCS. If this hypothesis is correct, the composition of bubbles that develop during decompression should reflect such gas differences. Unanesthetized guinea pigs were compressed to depths ranging from 250 to 350 fsw with air, He-O2 (21% O2) or one of a number of N2-He-O2 or N2-Ar-O2 mixtures (21% O2). Animals were held at depth from 15 to 60 min, then decompressed slowly (60 fsw/min) or rapidly (less than 15 s) to 5 fsw. If severe DCS developed, as judged by changes in physiologic variables, death usually occurred quickly. Gas/blood samples were then immediately withdrawn from the right atrium or the inferior vena cava, and the gas phase analyzed for He, N2, Ar, O2, and CO2 via gas chromatography. Bubbles from all dives contained 5-9% CO2, 1-4% O2, with the balance inert gas. Bubbles after N2-He-O2 dives contained substantially more N2 than He (up to 1.9 times more) compared to the dive mixture; bubbles after N2-Ar-O2 dives contained more Ar than N2 (up to 1.8 times more). For N2-He-O2 dives, the actual inert gas makeup of bubbles was dependent on the time-at-depth and the decompression profile. Results may reflect differences among He, N2, and Ar in tissue solubility/diffusivity and gas exchange rates, and support the rank order of increasing DCS risk (He less than N2 less than Ar) and rate of gas exchange (N2 less than He) observed previously during rat dives.

Decompression comparison of N2 and O2 in rats.

We have previously reported that O2 in the breathing gas mixture contributed significantly to the risk of decompression sickness (DCS) in rats after rapid (less than 10 s) decompression to the surface from depth. The rate of O2 uptake was extremely fast (less than 1 min estimated for equilibrium after a pressure change) compared to much slower rates for He and N2. To further define the role that O2 plays in diving, the present investigation examined decompression outcome in unanesthetized male albino rats after 60-min N2-O2 dives (1-3 atm abs O2, depth 6.26 or 7.26 atm abs). Slower decompression profiles were used to determine the elimination rates of N2 and O2 as pressure was reduced and included "stops" of up to 20 min. The probability of DCS was modeled using the maximum likelihood technique. O2 again contributed significantly to the risk of DCS, although O2 was eliminated very rapidly during decompression; the washout of N2 was considerably longer. These findings support the view that O2 can add significantly to decompression risk. However, this phenomenon may not normally be encountered during human diving operations where relatively slower decompression and lower PO2's are used.

Atmosphere contamination following repainting of a human hyperbaric chamber complex.

The Naval Medical Research Institute currently conducts hyperbaric research in a Man-Rated Chamber Complex (MRCC) originally installed in 1977. Significant engineering alterations to the MRCC and rusting of some of its interior sections necessitated repainting, which was completed in 1988. Great care was taken in selecting an appropriate paint (polyamide epoxy) and in ensuring correct application and curing procedures. Only very low levels of hydrocarbons were found in the MRCC atmosphere before initial pressurization after painting and curing. After pressurization, however, significant chemical contamination was found. The primary contaminants were aromatic hydrocarbons: xylenes (which were a major component of both the primer and topcoat paint) and ethyl benzene. The role that pressure played in stimulating off-gassing from the paint is not clear; the off-gassing rate was observed to be similar over a large range in chamber pressures from 1.6 to 31.0 atm abs. Scrubbing the chamber atmosphere with the chemical absorbent Purafil was effective in removing the contaminants. Contamination has been observed to slowly decline with chamber use and is expected to continue to improve with time. However, this contamination experience emphasizes the need for a high precision gas analysis program at any diving facility to ensure the safety of the breathing gas and chamber atmosphere.

Effect of inert gas switching at depth on decompression outcome in rats.

The present investigation was performed to determine whether inert gas sequencing at depth would affect decompression outcome in rats via the phenomenon of counterdiffusion. Unanesthetized rats (Rattus norvegicus) were subjected to simulated dives in either air, 79% He-21% O2, or 79% Ar-21% O2; depths ranged from 125 to 175 feet of seawater (4.8-6.3 atmospheres absolute). After 1 h at depth, the dive chamber was vented (with depth held constant) over a 5-min period with the same gas as in the chamber (controls) or one of the other two inert gas-O2 mixtures. After the gas switch, a 5- to 35-min period was allowed for gas exchange between the animals and chamber atmosphere before rapid decompression to the surface. Substantial changes in the risk of decompression sickness (DCS) were observed after the gas switch because of differences in potencies (He less than N2 less than Ar) for causing DCS and gas exchange rates (He greater than Ar greater than N2) among the three gases. Based on the predicted gas exchange rates, transient increases or decreases in total inert gas pressure would be expected to occur during these experimental conditions. Because of differences in gas potencies, DCS risk may not directly follow the changes in total inert gas pressure. In fact, a decline in predicted DCS risk may occur even as total inert gas pressure in increasing.

Air vs. He-O2 recompression treatment of decompression sickness in guinea pigs.

Air vs. He-O2 (20.9% O2) recompression treatment was examined in a model of severe decompression sickness (DCS) using male albino guinea pigs (Cavia porcellus, 500-600 g). Following decompression to the surface from simulated air dives at 200 or 250 fsw, both anesthetized and unanesthetized animals often exhibited responses indicative of a fatal bout of DCS (including hypotension, cardiac arrhythmia, and tachypnea). Upon recompression with air back to depth, good recovery of animals with DCS was observed. Comparison of air vs. He-O2 recompression responses of unanesthetized animals with recompression back to initial depth (200 fsw) revealed a slower recovery from tachypnea with He-O2. Recompression partially back to depth following 200-fsw air dives produced significant differences in the breathing recovery vs. recompression depth relationship between air and He-O2. Treatment effectiveness improved with increasing depth with air, but not with He-O2. These data indicate potential differences in recompression response to air vs. He-O2 when using ventilatory recovery as a measure of effectiveness in treatment of DCS in guinea pigs following air dives.

Effect of N2-He-O2 on decompression outcome in rats after variable time-at-depth dives.

No study of decompression sickness has examined both variable gas mixtures and variable time at depth to the point of statistical significance. This investigation examined the effect of N2-He-O2 on decompression outcome in rats after variable time-at-depth dives. Unanesthetized male albino rats were subjected to one of two series of simulated dives: 1) N2-He-O2 dives (20.9% O2) at 175 feet of seawater fsw) and 2) N2-O2 dives (variable percentage of O2; depths from 141 to 207 fsw). Time at depth ranged from 10 to 120 min; rats were then decompressed within 10 s to surface pressure. The probability of decompression sickness (severe bends symptoms or death) was analyzed with a Hill equation model, with parameters for gas potency and equilibrium time for the three gases and weight of the animal. Relative potencies for the three gases were of similar magnitude for bends and statistically different for death in ascending order: O2 less than He less than N2. Estimated gas uptake rates were different. N2 took three to four times as long as He to reach full effect; the rate of O2 appeared to be considerably shorter than that of N2 or He. The large influence of O2 on decompression outcome questions the simplistic view that O2 cannot contribute to the decompression requirement.

Decompression outcome following saturation dives with multiple inert gases in rats.

This investigation examined the question of whether gas mixtures containing multiple inert gases provide a decompression advantage over mixtures containing a single inert gas. Unanesthetized male albino rats, Rattus norvegicus, were subjected to 2-h simulated dives at depths ranging from 145 to 220 fsw. At pressure, the rats breathed various He-N2-Ar-O2 mixtures (79.1% inert gas-20.9% O2); they were then decompressed rapidly (within 10 s) to surface pressures. The probability of decompression sickness (DCS), measured either as severe bends symptoms or death, was related to the experimental variables in a Hill equation model incorporating parameters that account for differences in the potencies of the three gases and the weight of the animal. The relative potencies of the three gases, which affect the total dose of decompression stress, were determined as significantly different in the following ascending order of potency: He less than N2 less than Ar; some of these differences were small in magnitude. With mixtures, the degree of decompression stress diminished as either N2 or Ar was replaced by He. No obvious advantage or disadvantage of mixtures over the least potent pure inert gas (He) was evident, although limits to the expectation of possible advantage or disadvantage of mixtures were defined. Also, model analysis did not support the hypothesis that the outcome of decompression with multiple inert gases in rats under these experimental conditions can be explained totally by the volume of gas accumulated in the body during a dive.

Influence of ischemia and hypoxia on breathing in ducks.

We examined the influence of whole and lower body hypoxia and lower body ischemia on breathing in White Pekin ducks, Anas platyrhynchos, excluding pathways involving the carotid bodies. Carotid body denervated birds breathing 10 or 5% O2 developed a tachypnea after a latency of 30-100 s. The tachypnea was more pronounced with the more severe hypoxia, resulting in almost a doubling of minute ventilation (VE). Occlusion of the abdominal aorta in unanesthetized ducks produced immediate development of hypertension. Ventilation was unaffected for the 1st min; a tachypnea then developed rapidly and persisted for the duration of the occlusion resulting in a 25% increase in VE. After thoracic spinal section, all ventilatory responses to occlusion were eliminated. Experimental perfusion of the brain and single intact carotid body in unanesthetized ducks with hyperoxic blood during low O2 breathing (6-9% O2) resulted in tachypnea, also after a considerable latency. These results suggest that severe hypoxia can affect breathing in birds via pathways other than those involving the carotid bodies.

Control of diving responses by carotid bodies and baroreceptors in ducks.

The precise role of carotid body chemoreceptors and systemic baroreceptors in cardiovascular responses during experimental diving in ducks is controversial. The diving responses of chronically baroreceptor-denervated, chemoreceptor-denervated, and combined baroreceptor- and chemoreceptor-denervated White Pekin ducks, Anas platyrhynchos, were compared with those of intact and sham-operated birds. All three types of denervation elevated predive heart rates on average by 100-150 beats/min. During submergence, the cardiac rate of the barodenervates quickly dropped and after 60 s stabilized at levels similar to those of submerged intact ducks for the remainder of a 2-min dive. However, arterial blood pressure declined drastically in the barodenervates. Ducks without functional carotid bodies showed significant bradycardia during submergence, although heart rate only fell to the predive rate of intact animals. Birds with combined baroreceptor and chemoreceptor denervation exhibited the same degree of bradycardia as chemoreceptor denervates, and arterial blood pressure rose spectacularly during a dive. It is concluded that during experimental diving in ducks 1) cardiac responses are not baroreflexive in origin, 2) the major portion of bradycardia is due to stimulation of carotid body chemoreceptors, and 3) intact system baroreceptors appear essential for maintenance of blood pressure.

Effect of cardiovascular variables on hyperpnea during recovery from diving in ducks.

Control of hyperpnea during recovery from diving in unanesthetized White Pekin ducks, Anas platyrhynchos, was examined. Postdive minute ventilation (VE) increased five times regardless of the length of the preceding dive (1-4 min), although longer dives resulted in slower return of VE towards predive levels. Manipulation of arterial blood gases showed that both hypoxia and hypercapnia contributed to hyperpnea on emergence. Although chronic bilateral carotid body denervation depressed VE before and after diving, VE still increased four times after 1-min dives. Postdive hyperpnea was accompanied by dramatic elevations in heart rate, cardiac output, and the ventilation/perfusion ratio. However, artificially maintaining heart rate at abnormally low levels did not affect the postdive hyperpnea. In addition, postdive hyperpnea was unaffected by systemic arterial baroreceptor denervation. Postdive hyperpnea in ducks depends on blood gas changes occurring during a dive, yet a substantial part of the response is independent of input from carotid body chemoreceptors and the accompanying rises in heart rate and cardiac output.

Autonomic cardiovascular control during submergence and emergence in bullfrogs.

Unanesthetized bullfrogs were involuntarily submerged for 25 min in air-saturated water at 21 degrees C. Significant bradycardia was observed while systemic blood pressure was maintained or slightly elevated. Upon emergence, heart rates immediately returned to presubmergence levels or higher. Similar responses were observed in frogs allowed to make voluntary dives in an experimental tank. Heart rates of vagal-blocked (atropine) frogs did not change during submergence or emergence. beta-Adrenergic blockade (propranolol) had little effect on the magnitude of heart rate decrease during submergence or its increase upon emergence. After alpha-adrenergic blockade (phentolamine), frogs developed diving bradycardia while undergoing a fall in systemic blood pressure. It is concluded that, in bullfrogs, 1) bradycardia during submergence is entirely due to increased vagal activity, 2) the immediate cardiac rate increase upon emergence apparently results from a decrease in vagal tone; and 3) there appears to be no substantial reciprocal sympathetic influence on heart rate during alterations in vagal tone.

The depressant influence of extracellular fluid hyperoxia on liver slice oxygen uptake.

The effect of oxygen tension (Po2) on oxygen consumption (Vo2) of rabbit liver slices was investigated to determine the relationship between extracellular fluid Po2 and liver slice Vo2. Seventy rabbits, 5 kg each, were sacrificed with air embolism. Liver slices (0.3 gm) in Krebs-Ringer's-phosphate (KRP) dextrose were placed in a constantly agitated, 37 degrees C, closed cuvette, and Po2 was continuously monitored down to 10 torr. The system was reoxygenated and closed, and observations were repeated. Hemoglobin concentration was measured on the liver slice homogenate, and a P50 was measured on autologous blood. The presence of small amounts of hemoglobin in the supernatant was confirmed by electrophoretic, spectrophotometric, and morphologic studies. The pH decreased from mean 7.47 to mean 7.37 during each run. The resultant increase in P50 was recalculated and included in the determination of each deltaO2 content/min (Vo2). Vo2 at Po2 30 torr was greater than Vo2 at Po2 90 torr (p less than 0.0001 in each case). Vo2 at Po2 30 torr was not significantly different in the first vs. the second run (p greater than 0.85). The critical oxygen tension for hepatocyte respiration appears to be 30 torr.

Direct influence of endotoxin on cellular respiration.

Hepatocyte oxygen consumption was evaluated in vitro in 70 rabbits. In selected instances, Escherichia coli endotoxin was added to a chamber containing either rabbit hepatocytes or hepatocyte homogenates. Endotoxin directly depressed the respiration of intact rabbit hepatocytes. Endotoxin also decreased the oxygen consumption of hepatocyte homogenates. The intact cell membrane is not an effective barrier to the deleterious effect of endotoxin. A patient in a state of endotoxic shock, in addition to having a hemodynamic derangement, probably has circulating toxins that directly render cellular oxidative phsophorylation less efficient.