Acute and potentially persistent effects of scuba diving on the blood transcriptome of experienced divers.

During scuba diving, the circulatory system is stressed by an elevated partial pressure of oxygen while the diver is submerged and by decompression-induced gas bubbles on ascent to the surface. This diving-induced stress may trigger decompression illness, but the majority of dives are asymptomatic. In this study we have mapped divers' blood transcriptomes with the aim of identifying genes, biological pathways, and cell types perturbed by the physiological stress in asymptomatic scuba diving. Ten experienced divers abstained from diving for >2 wk before performing a 3-day series of daily dives to 18 m depth for 47 min while breathing compressed air. Blood for microarray analysis was collected before and immediately after the first and last dives, and 10 matched nondivers provided controls for predive stationary transcriptomes. MetaCore GeneGo analysis of the predive samples identified stationary upregulation of genes associated with apoptosis, inflammation, and innate immune responses in the divers, most significantly involving genes in the TNFR1 pathway of caspase-dependent apoptosis, HSP60/HSP70 signaling via TLR4, and NF-κB-mediated transcription. Diving caused pronounced shifts in transcription patterns characteristic of specific leukocytes, with downregulation of genes expressed by CD8+ T lymphocytes and NK cells and upregulation of genes expressed by neutrophils, monocytes, and macrophages. Antioxidant genes were upregulated. Similar transient responses were observed after the first and last dive. The results indicate that sublethal oxidative stress elicits the myeloid innate immune system in scuba diving and that extensive diving may cause persistent change in pathways controlling apoptosis, inflammation, and innate immune responses.

Differentiation at autopsy between in vivo gas embolism and putrefaction using gas composition analysis.

Gas embolism can arise from different causes (iatrogenic accidents, criminal interventions, or diving related accidents). Gas analyses have been shown to be a valid technique to differentiate between putrefaction gases and gas embolism. In this study, we performed systematic necropsies at different postmortem times in three experimental New Zealand White Rabbits models: control or putrefaction, infused air embolism, and compression/decompression. The purpose of this study was to look for qualitative and quantitative differences among groups and to observe how putrefaction gases mask in vivo gas embolism. We found that the infused air embolism and compression/decompression models had a similar gas composition prior to 27-h postmortem, being typically composed of around 70-80 % of N(2) and 20-30 % of CO(2), although unexpected higher CO(2) concentrations were found in some decompressed animals, putting in question the role of CO(2) in decompression. All these samples were statistically and significantly different from more decomposed samples. Gas composition of samples from more decomposed animals and from the putrefaction model presented hydrogen, which was therefore considered as a putrefaction marker.

Simulated dive in rats lead to acute changes in cerebral blood flow on MRI, but no cerebral injuries to grey or white matter.

In this study, the effect of a simulated dive on rat brain was investigated using several magnetic resonance imaging (MRI)-methods and immunohistochemistry. Rats were randomly assigned to a dive- or a control group. The dive group was exposed to a simulated air dive to 600 kPa for 45 min. Pulmonary artery was monitored for vascular gas bubbles by ultrasound. MRI was performed 1 h after decompression and at one and 2 weeks after the dive with a different combination of MRI sequences at each time point. Two weeks after decompression, rats were sacrificed and brains were prepared for histology. Dived rats had a different time-curve for the dynamic contrast-enhanced MRI signal than controls with higher relative signal intensity, a tendency towards longer time to peak and a larger area under the curve for the whole brain on the acute MRI scan. On MRI, 1 and 2 weeks after dive, T2-maps showed no signal abnormalities or morphological changes. However, region of interest based measurements of T2 showed higher T2 in the brain stem among decompressed animals than controls after one and 2 weeks. Microscopical examination including immunohistochemistry did not reveal apparent structural or cellular injuries in any part of the rat brains. These observations indicate that severe decompression does not seem to cause any structural or cellular injury to the brain tissue of the rat, but may cause circulatory changes in the brain perfusion in the acute phase.

Exercise-induced myofibrillar disruption with sarcolemmal integrity prior to simulated diving has no effect on vascular bubble formation in rats.

Decompression sickness is initiated by gas bubbles formed during decompression, and it has been generally accepted that exercise before decompression causes increased bubble formation. There are indications that exercise-induced muscle injury seems to be involved. Trauma-induced skeletal muscle injury and vigorous exercise that could theoretically injure muscle tissues before decompression have each been shown to result in profuse bubble formation. Based on these findings, we hypothesized that exercise-induced skeletal muscle injury prior to decompression from diving would cause increase of vascular bubbles and lower survival rates after decompression. In this study, we examined muscle injury caused by eccentric exercise in rats prior to simulated diving and we observed the resulting bubble formation. Female Sprague-Dawley rats (n = 42) ran downhill (-16º) for 100 min on a treadmill followed by 90 min rest before a 50-min simulated saturation dive (709 kPa) in a pressure chamber. Muscle injury was evaluated by immunohistochemistry and qPCR, and vascular bubbles after diving were detected by ultrasonic imaging. The exercise protocol resulted in increased mRNA expression of markers of muscle injury; αB-crystallin, NF-κB, and TNF-α, and myofibrillar disruption with preserved sarcolemmal integrity. Despite evident myofibrillar disruption after eccentric exercise, no differences in bubble amounts or survival rates were observed in the exercised animals as compared to non-exercised animals after diving, a novel finding that may be applicable to humans.

Early genetic responses in rat vascular tissue after simulated diving.

Diving causes a transient reduction of vascular function, but the mechanisms behind this are largely unknown. The aim of this study was therefore to analyze genetic reactions that may be involved in acute changes of vascular function in divers. Rats were exposed to 709 kPa of hyperbaric air (149 kPa Po(2)) for 50 min followed by postdive monitoring of vascular bubble formation and full genome microarray analysis of the aorta from diving rats (n = 8) and unexposed controls (n = 9). Upregulation of 23 genes was observed 1 h after simulated diving. The differential gene expression was characteristic of cellular responses to oxidative stress, with functions of upregulated genes including activation and fine-tuning of stress-responsive transcription, cytokine/cytokine receptor signaling, molecular chaperoning, and coagulation. By qRT-PCR, we verified increased transcription of neuron-derived orphan receptor-1 (Nr4a3), plasminogen activator inhibitor 1 (Serpine1), cytokine TWEAK receptor FN14 (Tnfrsf12a), transcription factor class E basic helix-loop-helix protein 40 (Bhlhe40), and adrenomedullin (Adm). Hypoxia-inducible transcription factor HIF1 subunit HIF1-α was stabilized in the aorta 1 h after diving, and after 4 h there was a fivefold increase in total protein levels of the procoagulant plasminogen activator inhibitor 1 (PAI1) in blood plasma from diving rats. The study did not have sufficient power for individual assessment of effects of hyperoxia and decompression-induced bubbles on postdive gene expression. However, differential gene expression in rats without venous bubbles was similar to that of all the diving rats, indicating that elevated Po(2) instigated the observed genetic reactions.

Concentration of circulating autoantibodies against HSP 60 is lowered through diving when compared to non-diving rats.

OBJECTIVE: Skin and ear infections, primarily caused by Pseudomonas aeruginosa (P. aeruginosa), are recurrent problems for saturation divers, whereas infections caused by P. aeruginosa are seldom observed in healthy people outside saturation chambers. Cystic fibrosis (CF) patients suffer from pulmonary infections by P. aeruginosa, and it has been demonstrated that CF patients have high levels of autoantibodies against Heat shock protein 60 (HSP60) compared to controls, probably due to cross-reacting antibodies induced by P. aeruginosa. The present study investigated whether rats immunised with P. aeruginosa produced autoantibodies against their own HSP60 and whether diving influenced the level of circulating anti-HSP60 antibodies. METHODS: A total of 24 rats were randomly assigned to one of three groups ('immunised', 'dived' and 'immunised and dived'). The rats in group 1 and 3 were immunised with the bacteria P. aeruginosa, every other week. Groups 2 and 3 were exposed to simulated air dives to 400 kPa (4 ata) with 45 min bottom time, every week for 7 weeks. Immediately after surfacing, the rats were anaesthetised and blood was collected from the saphenous vein. The amount of anti-HSP60 rat antibodies in the serum was analysed by enzyme linked immunosorbent assay. RESULTS: The immunised rats (group 1) showed a significant increase in the level of autoantibodies against HSP60, whereas no autoantibodies were detected in the dived rats (group 2). The rats both immunised and dived (group 3) show no significant increase in circulating autoantibodies against HSP60. A possible explanation may be that HSP60 is expressed during diving and that cross-reacting antibodies are bound.

Use of heart rate monitoring for an individualized and time-variant decompression model.

Individual differences, physiological pre-conditions and in-dive conditions like workload and body temperature have been known to influence bubble formation and risk of decompression sickness in diving. Despite this fact, such effects are currently omitted from the decompression algorithms and tables that are aiding the divers. There is an apparent need to expand the modeling beyond depth and time to increase safety and efficiency of diving. The present paper outlines a mathematical model for how heart rate monitoring in combination with individual parameters can be used to obtain a customized and time-variant decompression model. We suggest that this can cover some of the individual differences and dive conditions that are affecting bubble formation. The model is demonstrated in combination with the previously published Copernicus decompression model, and is suitable for implementation in dive computers and post dive simulation software for more accurate risk analysis.

A reliable and valid protocol for measuring maximal oxygen uptake in pigs.

BACKGROUND: Most knowledge about cellular and molecular adaptation in the heart after exercise training comes from rodent models, and this has substantially improved our knowledge about exercise-induced cardiac adaptations. However, in rodents, the electrophysiological properties of the heart are different from the human heart. Therefore, the need of exercise-training models in larger animal models is obvious. Physiological studies of cardio-respiratory fitness require training regimens that give robust and adequate testing procedures to quantify the outcome. METHODS: We developed a valid and reproducible protocol for measuring maximal oxygen uptake (VO2max) in young pigs. As previous studies have exercised pigs using horizontal treadmills, we determined whether treadmill inclination may influence the level of peak oxygen uptake (VO2peak) achieved, and whether the true VO2max was reached. Eight young pigs were used. Submaximal and VO2peak were tested at five different inclinations from 13 to 30 degrees . RESULTS: At submaximal VO2, there was an excellent test-retest at all treadmill inclinations (r = 0.99, coefficient of variation = 1.8%). The level of VO2peak was dependent upon treadmill inclination and the true VO2max, defined as a levelling-off of VO2 despite increased running speed, was only reached a treadmill inclination of 24 degrees . For VO2peak we only observed a significant test-retest correlation when using 19 and 24 degrees inclination of the treadmill (r = 0.88, coefficient of variation = 9.7%). CONCLUSION: The use of inappropriate treadmill inclination might hide training-induced adaptations if the true VO2max is not reached. This study shows that the present test protocol can be used in future studies of exercise on treadmill, when the aim is to measure submaximal and VO2max in pigs.

Expression of endothelial selectin ligands on human leukocytes following dive.

The fact that impaired endothelial-dependent vasodilatation after scuba diving often occurs without visible changes in the endothelial layer implies its biochemical origin. Since Lewisx(CD15) and sialyl-Lewisx(CD15s) are granulocyte and monocyte carbohydrate antigens recognized as ligands by endothelial selectins, we assumed that they could be sensitive markers for impaired vasodilatation following diving. Using flow cytometry, we determined the CD15 and CD15s peripheral blood mononuclear cells of eight divers, 30 mins before and 50 mins after a single dive to 54 m for 20 mins bottom time. The number of gas bubbles in the right heart was monitored by ultrasound. Gas bubbles were seen in all eight divers, with the average number of bubbles/cm2 1.9+/-1.9. The proportion of CD15+monocytes increased 2-fold after the dive as well as the subpopulation of monocytes highly expressing CD15s. The absolute number of monocytes was slightly, but not significantly, increased after the dive, whereas the absolute number of granulocytes was markedly elevated (up to 61%). There were no significant correlations between bubble formation and CD15+monocyte expression (r=-0.56; P=0.17), as well as with monocytes highly expressing CD15s (r=0.43; P=0.29). This study suggests that biochemical changes induced by scuba diving primarily activate existing monocytes rather than increase the number of monocytes at a time of acute arterial endothelial dysfunction.

High-grade bubbles in left and right heart in an asymptomatic diver at rest after surfacing.

INTRODUCTION: Most decompression procedures induce the formation of asymptomatic venous gas bubbles. They can be classified as "silent bubbles," which are asymptomatic compared to paradoxical arterialization of venous gas emboli, which can lead to serious neurologic damage. The penetration of such gas bubbles into the arterial circulation is due to pulmonary barotrauma, intrapulmonary (I-P) passage after massive bubble formation ("chokes"), or intracardiac shunting. Venous gas bubbles can be monitored and graded with echocardiographic scanning. CASE: We believe this is the first case to be reported of a recreational diver who, after surfacing from a dive, developed grade 5 ("white-out") venous gas bubbles in the right heart with evidence of I-P shunt at rest without any symptoms of decompression sickness. Grade 4 gas bubbles were found on the left side of the heart, indicating significant I-P shunting even at rest. CONCLUSION: We observed venous bubbles crossing through the I-P shunt during post-dive recovery at rest in a diver who developed "white out" of venous bubbles. Previously, the maximum bubble grade 5 had been observed in experimental animals, but not in humans. Moreover, a significant bubble grade was found on the left side of the heart, indicating a need for further studies to investigate the mechanisms of post-dive changes in peripheral and central circulation.

Validation of decompression procedures based on detection of venous gas bubbles: A Bayesian approach.

INTRODUCTION: Verification of new decompression procedures has traditionally been based on observing the occurrence of decompression sickness (DCS) in test dives. Several hundred exposures are required to determine the safety of a procedure with any degree of certainty. The number of venous gas emboli (VGE) corresponds with the risk of getting DCS and detection of VGE has been used as an alternative method for validation of decompression procedures. We propose a new and improved method for validation based on detection of VGE. METHODS: Our Bayesian statistics method combines results from ultrasound detection of VCE in test dives with knowledge about the correspondence between VGE and DCS risk obtained from a large number of previous experimental studies. Our algorithm is implemented in a computer program; it estimates DCS risk and 95% credible intervals for the tested procedure. RESULTS: We have applied the method to available VGE data from tested air diving procedures with between 7 and 14 test dives for each procedure. The estimated credible intervals correspond to confidence intervals from 130-250 dives using the binomial distribution of the traditional "DCS observation validation." DISCUSSION: We conclude that, compared with previous methods, the proposed method can greatly reduce the number of dives required to validate or reject new decompression procedures.

Recompression during decompression and effects on bubble formation in the pig.

INTRODUCTION: There is a relationship between gas bubble formation in the vascular system and serious decompression sickness. Hence, control of the formation of vascular bubbles should allow safer decompression procedures. METHODS: There were 12 pigs that were randomly divided into an experimental group (EXP) and a control group (CTR) of 6 animals each. The pigs were compressed to 500 kPa (5 ATA) in a dry hyperbaric chamber and held for 90 min bottom time breathing air. CTR animals were decompressed according to a modified USN dive profile requiring four stops. EXP followed the same profile except that a 5-min recompression of 50 kPa (0.5 ATA) was added at the end of each of the last three decompression stops before ascending to the next stop depth. RESULTS: All CTR animals developed bubbles, compared with only one animal in EXP. The number of bubbles detected during and after the dive was 0.02 +/- 0.02 bubbles x cm(-2) in CTR, while the number of bubbles detected in EXP were 0.0009 +/- 0.005 bubbles x cm(-2); the difference was highly significant. CONCLUSION: By brief recompression during late decompression stops, the amount of bubbles was reduced. Our findings give further support for a gas phase model of decompression.

Antioxidant pretreatment and reduced arterial endothelial dysfunction after diving.

INTRODUCTION: We have recently shown that a single air dive leads to acute arterial vasodilation and impairment of endothelium-dependent vasodilatation in humans. Additionally we have found that predive antioxidants at the upper recommended daily allowance partially prevented some of the negative effects of the dive. In this study we prospectively evaluated the effect of long-term antioxidants at a lower RDA dose on arterial endothelial function. METHODS: Eight professional male divers performed an open sea air dive to 30 msw. Brachial artery flow-mediated dilation (FMD) was assessed before and after diving. RESULTS: The first dive, without antioxidants, caused significant brachial arterial diameter increase from 3.85 +/- 0.55 to 4.04 +/- 0.5 mm and a significant reduction of FMD from 7.6 +/- 2.7 to 2.8 +/- 2.1%. The second dive, with antioxidants, showed unchanged arterial diameter and significant reduction of FMD from 8.11 +/- 2.4 to 6.8 +/- 1.4%. The FMD reduction was significantly less with antioxidants. Vascular smooth muscle function, assessed by nitroglycerine (endothelium-independent dilation), was unaffected by diving. DISCUSSION: This study shows that long-term antioxidant treatment at a lower RDA dose ending 3-4 h before a dive reduces the endothelial dysfunction in divers. Since the scuba dive was of a similar depth and duration to those practiced by numerous recreational divers, this study raises the possibility of routine predive supplementation with antioxidants.

Exogenous nitric oxide and bubble formation in divers.

PURPOSE: Prevention of bubble formation is a central goal in standard decompression procedures. Previously we have shown that exercise 20-24 h prior to a dive reduces bubble formation and increases survival in rats exposed to a simulated dive. Furthermore, we have demonstrated that nitric oxide (NO) may be involved in this protection; blocking the production of NO increases bubble formation while giving rats a long-lasting NO donor 20 h and immediately prior to a dive reduces bubble formation. This study determined whether a short-lasting NO donor, nitroglycerine, reduced bubble formation after standard dives and decompression in man. METHODS: A total of 16 experienced divers were randomly assigned into two groups. One group performed two dives to 30 m of seawater (msw) for 30 min breathing air, and performed exercise at an intensity corresponding to 30% of maximal oxygen uptake during the bottom time. The second group performed two simulated dives to 18 msw for 80 min breathing air in a hyperbaric chamber, and remained sedentary during the bottom period. The first dive for each diver served as the control dive, whereas the divers received 0.4 mg of nitroglycerine by oral spray 30 min before the second dive. Following the dive, gas bubbles in the pulmonary artery were recorded using ultrasound. RESULTS: The open-water dive resulted in significantly more gas bubbles than the dry dive (0.87 +/- 1.3 vs 0.12 +/- 0.23 bubbles per square centimeter). Nitroglycerine reduced bubble formation significantly in both dives from 0.87 +/- 1.3 to 0.32 +/- 0.7 in the in-water dive and from 0.12 +/- 0.23 to 0.03 +/- 0.03 bubbles per square centimeter in the chamber dive. CONCLUSION: The present study demonstrates that intake of a short-lasting NO donor reduces bubble formation following decompression after different dives.

Post-dive bubble formation in rats: effects of exercise 24 h ahead repeated 30 min before the dive.

INTRODUCTION: Recent studies have shown that a nitric oxide releasing agent or a single bout of high-intensity exercise 20-24 h before a dive can prevent bubble formation following decompression. The aim of this study was to determine whether high-intensity exercise immediately prior to a dive eliminates the protective effect of a single bout of high-intensity exercise 24 h before the dive. METHODS: Twelve female Sprague-Dawley rats were randomly divided into two equal groups. Group 1 performed 90 min of exercise twice, beginning 24.5 h and again 2.0 h before compression. Group 2 performed 90 min of exercise beginning at 25.5 h before compression. The standardized exercise protocol was 7 x 8 min at 85-90% maximal oxygen uptake (Vo2max) followed by 2 min at 50% Vo2max for a total of 90 min including a 20 min warm-up at 40-50% of Vo2max. All rats were exposed to a pressure of 700 kPa (7 ATA) for 45 min in a dry hyperbaric chamber followed by decompression to the surface at 100 kPa (1 ATA) at a rate of 50 kPa x min(-1) (0.5 atm x min(-1)) breathing air. RESULTS: Bubble formation was significantly higher in rats that had exercised 24 h and 30 min prior to dive than rats that had only exercised 24 h prior to the dive (median bubble grade 4.5 vs. 0.5). CONCLUSION: This study demonstrated that acute exercise prior to a dive eliminated the protection against bubble formation found 24 h after high-intensity exercise in rats.

Bubbles Quantified In vivo by Ultrasound Relates to Amount of Gas Detected Post-mortem in Rabbits Decompressed from High Pressure.

The pathophysiological mechanism of decompression sickness is not fully understood but there is evidence that it can be caused by intravascular and autochthonous bubbles. Doppler ultrasound at a given circulatory location is used to detect and quantify the presence of intravascular gas bubbles as an indicator of decompression stress. In this manuscript we studied the relationship between presence and quantity of gas bubbles by echosonography of the pulmonary artery of anesthetized, air-breathing New Zealand White rabbits that were compressed and decompressed. Mortality rate, presence, quantity, and distribution of gas bubbles elsewhere in the body was examined postmortem. We found a strong positive relationship between high ultrasound bubble grades in the pulmonary artery, sudden death, and high amount of intra and extra vascular gas bubbles widespread throughout the entire organism. In contrast, animals with lower bubble grades survived for 1 h after decompression until sacrificed, and showed no gas bubbles during dissection.

Differentiation at necropsy between in vivo gas embolism and putrefaction using a gas score.

Gas bubble lesions consistent with decompression sickness in marine mammals were described for the first time in beaked whales stranded in temporal and spatial association with military exercises. Putrefaction gas is a post-mortem artifact, which hinders the interpretation of gas found at necropsy. Gas analyses have been proven to help differentiating putrefaction gases from gases formed after hyperbaric exposures. Unfortunately, chemical analysis cannot always be performed. Post-mortem computed tomography is used to study gas collections, but many different logistical obstacles and obvious challenges, like the size of the animal or the transport of the animal from the stranding location to the scanner, limit its use in stranded marine mammals. In this study, we tested the diagnostic value of an index-based method for characterizing the amount and topography of gas found grossly during necropsies. For this purpose, putrefaction gases, intravenously infused atmospheric air, and gases produced by decompression were evaluated at necropsy with increased post-mortem time in New Zealand White Rabbits using a gas score index. Statistical differences (P<0.001) were found between the three experimental models immediately after death. Differences in gas score between in vivo gas embolism and putrefaction gases were found significant (P<0.05) throughout the 67h post-mortem. The gas score-index is a new and simple method that can be used by all stranding networks, which has been shown through this study to be a valid diagnostic tool to distinguish between fatal decompression, iatrogenic air embolism and putrefaction gases at autopsies.

Exercise-induced intrapulmonary shunting of venous gas emboli does not occur after open-sea diving.

Paradoxical arterializations of venous gas emboli can lead to neurological damage after diving with compressed air. Recently, significant exercise-induced intrapulmonary anatomical shunts have been reported in healthy humans that result in widening of alveolar-to-arterial oxygen gradient. The aim of this study was to examine whether intrapulmonary shunts can be found following strenuous exercise after diving and, if so, whether exercise should be avoided during that period. Eleven healthy, military male divers performed an open-sea dive to 30 m breathing air, remaining at pressure for 30 min. During the bottom phase of the dive, subjects performed mild exercise at approximately 30% of their maximal oxygen uptake. The ascent rate was 9 m/min. Each diver performed graded upright cycle ergometry up to 80% of the maximal oxygen uptake 40 min after the dive. Monitoring of venous gas emboli was performed in both the right and left heart with an ultrasonic scanner every 20 min for 60 min after reaching the surface pressure during supine rest and following two coughs. The diving profile used in this study produced significant amounts of venous bubbles. No evidence of intrapulmonary shunting was found in any subject during either supine resting posture or any exercise grade. Also, short strenuous exercise after the dive did not result in delayed-onset decompression sickness in any subject, but studies with a greater number of participants are needed to confirm whether divers should be allowed to exercise after diving.

Exercise ending 30 min pre-dive has no effect on bubble formation in the rat.

INTRODUCTION: We have previously shown that exercise performed 20 h before a dive significantly reduces bubble formation in both rats and humans. Furthermore, exercise performed closer to the dive did not prevent bubble formation. HYPOTHESIS: The present study was designed to determine whether exercise 30 min prior to a dive promotes bubble formation. The occurrence of many bubbles is linked to a higher risk of developing decompression sickness. METHODS: A total of 58 Sprague-Dawley rats were randomly divided into a sedentary control group (n = 29) and an exercise group (n = 29). Rats in the exercise group ran on a treadmill for a total of 90 min at variable intensity up to 85-90% of VO2max. Then, 30 min after exercise, one rat from each group rested in a pressure chamber at 700 kPa (7 atm) breathing air, performing a simulated dive. Bottom time was 45 min; decompression rate was 50 kPa x min(-1) (0.5 atm x min(-1)). Immediately after surfacing (100 kPa, 1 atm), the rats were anesthetized and bubbles were measured discontinuously for the next 60 min. RESULTS: There were no significant differences in survival (p = 0.55), median bubble grade (p = 0.67), survival time (p = 0.53), or the number of rats getting a bubble score > or = 2 (p = 0.79) between the groups. CONCLUSION: The same type and intensity of exercise that reduces bubble formation when performed 20 h prior to a dive neither promotes nor reduces bubble formation if performed 30 min before a dive. The present data indicate that exercise completed 30 min before a dive does not increase the risk of developing decompression sickness in the rat.

Career perspective: Alf O. Brubakk-looking back to see ahead.

The following describes my professional life up till today, but it also describes what I think lies ahead. I have led an interesting professional life and been lucky enough to be at the centre of some of the important development in modern medicine and diving, namely ultrasound in cardiology and the mechanisms of decompression. I therefore should be able to see some of the most challenging and exciting problems ahead. Ultrasound in cardiology has developed from simply listening to the Doppler signal to determine the velocity of blood flow to the complicated description of images presented today. Diving, in addition to being an important commercial and environmental activity, exposes the individual to intermittent hyperoxia and pressure reductions. These challenges evoke the production of radical oxygen species (ROS) and microparticles (MP) that also are central to many pathophysiologic mechanisms that are involved in a number of severe human diseases. Thus, diving can be regarded as an important model of disease and allows us to study their effects on healthy young individuals. The future thus points towards an integration of environmental physiology with detailed physiological and pathophysiological mechanisms and makes diving physiology a potentially very important field of study.