Andreas Møllerløkken

Venous and Arterial Bubbles at Rest after No-Decompression Air Dives

PURPOSE During SCUBA diving, breathing at increased pressure leads to a greater tissue gas uptake. During ascent, tissues may become supersaturated, and the gas is released in the form of bubbles that typically occur on the venous side of circulation. These venous gas emboli (VGE) are usually eliminated as they pass through the lungs, although their occasional presence in systemic circulation (arterialization) has been reported and it was assumed to be the main cause of the decompression sickness. The aims of the present study were to assess the appearance of VGE after air dives where no stops in coming to the surface are required and to assess their potential occurrence and frequency in the systemic circulation. METHODS Twelve male divers performed six dives with 3 d of rest between them following standard no-decompression dive procedures: 18/60, 18/70, 24/30, 24/40, 33/15, and 33/20 (the first value indicates depth in meters of sea water and the second value indicates bottom time in minutes). VGE monitoring was performed ultrasonographically every 20 min for 120 min after surfacing. RESULTS Diving profiles used in this study produced unexpectedly high amounts of gas bubbles, with most dives resulting in grade 4 (55/69 dives) on the bubble scale of 0-5 (no to maximal bubbles). Arterializations of gas bubbles were found in 5 (41.7%) of 12 divers and after 11 (16%) of 69 dives. These VGE crossovers were only observed when a large amount of bubbles was concomitantly present in the right valve of the heart. CONCLUSIONS Our findings indicate high amounts of gas bubbles produced after no-decompression air dives based on standardized diving protocols. High bubble loads were frequently associated with the crossover of VGE to the systemic circulation. Despite these findings, no acute decompression-related pathology was detected.

Venous gas embolism as a predictive tool for improving CNS decompression safety

A key process in the pathophysiological steps leading to decompression sickness (DCS) is the formation of inert gas bubbles. The adverse effects of decompression are still not fully understood, but it seems reasonable to suggest that the formation of venous gas emboli (VGE) and their effects on the endothelium may be the central mechanism leading to central nervous system (CNS) damage. Hence, VGE might also have impact on the long-term health effects of diving. In the present review, we highlight the findings from our laboratory related to the hypothesis that VGE formation is the main mechanism behind serious decompression injuries. In recent studies, we have determined the impact of VGE on endothelial function in both laboratory animals and in humans. We observed that the damage to the endothelium due to VGE was dose dependent, and that the amount of VGE can be affected both by aerobic exercise and exogenous nitric oxide (NO) intervention prior to a dive. We observed that NO reduced VGE during decompression, and pharmacological blocking of NO production increased VGE formation following a dive. The importance of micro-nuclei for the formation of VGE and how it can be possible to manipulate the formation of VGE are discussed together with the effects of VGE on the organism. In the last part of the review we introduce our thoughts for the future, and how the enigma of DCS should be approached.

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.

S100B and NSE serum concentrations after simulated diving in rats

The purpose of this study was to assess whether one could detect S100 calcium‐binding protein B (S100B) and neuron‐specific enolase (NSE) in serum of rats after a simulated dive breathing air, with the main hypothesis that the serum concentrations of S100B and NSE in rats will increase above pre‐exposure levels following severe decompression stress measured as venous gas emboli (VGE). The dive group was exposed to a simulated air dive to 700 kPa for 45 min. Pulmonary artery was monitored for vascular gas bubbles by ultrasound. Pre‐ and postdive blood samples were analyzed for S100B and NSE using commercially available Elisa kits. There was no increase in serum S100B or NSE after simulated diving and few of the animals were showing high bubble grades after the dives. The present study examined whether the protein biomarkers S100B and NSE could be found in serum from rats after exposure to a simulated dive to 700 kPa for 45 min breathing air. There were no differences in serum concentrations before versus after the dive exposure. This may be explained by the lack of vascular gas bubbles after the dives.

Aortic Function in Rats After Decompression Without Ultrasonically Detectable Bubble Formation

BACKGROUND Several studies have demonstrated an adverse effect of bubbles on endothelial function. The degree of dysfunction appears to be related to the number of bubbles present. The aim of the study was to determine whether decompression without bubble formation visible by ultrasound had any effect on arterial endothelial function. METHODS We decompressed 21 Sprague-Dawley rats weighing 215-260 g from 700 kPa (approximately 6.9 ATA) in a dry hyperbaric chamber followed by a 1-h observation period and measured aortic endothelial-dependent relaxation to acetylcholine. Later, we determined the specific weight of the brain as a measure of edema formation and vascular bubbles in the arterial circulation. RESULTS No bubbles were seen in the pulmonary arteries of seven rats. We found a significant lower vasodilatory response to acetylcholine in the decompressed rats (44% +/- 14%) compared to the control rats (58% +/- 12%) as a sign of endothelial dysfunction. There was no significant difference between the two groups in the specific gravity of the brain. CONCLUSION We conclude that measurable arterial dysfunction in the aorta can occur even if no visible venous bubble formation is seen. There are no results in this study suggesting that these rats had damaged blood-brain barriers or brain edema.

Blood gene expression and vascular function biomarkers in professional saturation diving

Saturation diving is an established way to conduct subsea operations with human intervention. While working, the divers must acclimatize to the hyperbaric environments. In this study, genome-wide gene expression and selected plasma biomarkers for vascular function were investigated. We also examined whether antioxidant vitamin supplements affected the outcome. The study included 20 male professional divers, 13 of whom took vitamin C and E supplements in doses of 1,000 and 30 mg daily during saturation periods that lasted 7–14 days. The dives were done in a heliox atmosphere with 40 kPa oxygen partial pressure (ppO2) to a depth of 100–115 m of sea-water (msw), from which the divers performed in-water work excursions to a maximum depth of 125 msw with 60 kPa ppO2. Venous blood was collected immediately before and after saturation. Following gene expression profiling, post-saturation gene activity changes were analyzed. Protein biomarkers for inflammation, endothelial function, and fibrinolysis: Il-6, CRP, ICAM-1, fibrinogen, and PAI-1, were measured in plasma. Post-saturation gene expression changes indicated acclimatization to elevated ppO2 by extensive downregulation of factors involved in oxygen transport, including heme, hemoglobin, and erythrocytes. Primary endogenous antioxidants; superoxide dismutase 1, catalase, and glutathione synthetase, were upregulated, and there was increased expression of genes involved in immune activity and inflammatory signaling pathways. The antioxidant vitamin supplements had no effect on post-saturation gene expression profiles or vascular function biomarkers, implying that the divers preserved their homeostasis through endogenous antioxidant defenses.

Diving and long-term cardiovascular health

Abstract Background Short-term cardiovascular effects from ambient pressure exposure are known. However, long-term cardiovascular effects from diving in humans have been less studied. Aims To examine possible long-term cardiovascular health effects from occupational diving. Methods We compared the prevalence of cardiovascular disease in former divers to non-divers. We obtained data on male former divers with a certificate valid for professional diving after 1980, from the Norwegian Diver 2011 project, and matched data on the general male population from the HUNT3 Survey. We also compared former divers with high and low grades of diving exposure. Results Data were available on 768 former divers. The prevalence of self-reported high blood pressure in former divers who often omitted a dive-free day after 3 days of strenuous diving was 28% compared with 18% in those who rarely violated these regulations [relative risk (RR) 1.47, confidence interval (CI) 1.01–2.15]. Also, the prevalence of myocardial infarction/angina pectoris was 11% in divers with >150 professional dives/year compared with 4% in divers with ≤50 professional dives/year [RR adj. 2.91 (CI 1.23–6.87)] and 16% in divers with >2000 air dives in total relative to 3% in divers with ≤2000 dives [RR adj. 3.05 (CI 1.47–6.34)]. Conclusions The prevalence of some cardiovascular symptoms and diseases may be higher in male former divers than in the general population. Diving might have adverse long-term cardiovascular effects. Whether this is associated with diving per se or strenuous physical activity requires further studies.

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.

Evaluating PAI-1 as a biomarker for stress in diving: human serum total PAI-1 is unaltered after 2 h dry exposures to 280 kPa hyperbaric air

Plasminogen activator inhibitor (PAI‐1) is induced in the vasculature and secreted into the vascular lumen in response to inflammation and oxidative stress. We have previously reported a fivefold increase in plasma PAI‐1 from rats exposed to 708 kPa hyperbaric air. In the current study we assess the potential of human serum total PAI‐1 as a biomarker for stress in compressed air diving. Eleven recreational divers, nine males and two females, completed four 2 h hyperbaric air exposures to 280 kPa in a pressure chamber over a period of 2 weeks. The air pressure corresponds to a diving depth of 18 m in water. Serum was collected before the study and again 3 h 30 min after completion of each hyperbaric exposure. All samples were taken in the afternoon to minimize the contribution of circadian variation. The analysis revealed no change in serum total PAI‐1 after hyperbaric exposures within the group of divers (P = 0.064), but significant interindividual differences persisted throughout the study (P < 0.0005). A case of decompression sickness after the third round of hyperbaric exposure did not affect PAI‐1. In conclusion, compressed air exposure to 280 kPa does not affect serum total PAI‐1, and significant interindividual variation in PAI‐1 levels may limit its usefulness as a biomarker. This does, however, not give a complete answer regarding PAI‐1 in physiologically stressful dives. Further studies with different exposures and timing are needed for that.

Erratum to: Venous gas embolism as a predictive tool for improving CNS decompression safety

Erratum to: Eur J Appl PhysiolDOI 10.1007/s00421-011-1998-9An incorrect citation was included in the originalpublication:Vince RV, Chrismas B, Midgley AW, McNaughton LR,Madden LA (2009) Hypoxia mediated release of endo-thelial microparticles and increased association ofS100A12 with circulating neutrophils. Oxid Med CellLongev 2:2–6The correct reference should be:Vince RV, McNaughton LR, Taylor L, Midgley AW,Laden G, Madden LA (2009) Release of VCAM-1 asso-ciated endothelial microparticles following simulatedSCUBA dives. Eur J Appl Physiol 105:507–513