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.(c) 2015 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited

Eccentric exercise 48 h prior to simulated diving has no effect on vascular bubble formation in rats

PURPOSE: Decompression sickness (DCS) caused by vascular bubble formation is a major risk when diving. Prior studies have shown that physical exercise has a significant impact in both reducing and increasing bubble formation. There is limited knowledge about the mechanisms, but there are indications that exercise-induced muscle injury prior to diving may cause increased bubble formation. The purpose of this study was to investigate the role of exercise-induced muscle injury as a possible mechanism of bubble formation during diving. METHODS: Muscle injury was induced by exposing female Sprague-Dawley rats (n = 30) to a single bout of eccentric exercise, 100 min intermittent, downhill (-16°) treadmill running. Forty-eight hours later, the animals were exposed to a 50-min simulated saturation dive (709 kPa) in a pressure chamber, when the degree of muscle injury and inflammation would be the most pronounced. Bubble formation after the dive was observed by ultrasonic imaging for 4 h. RESULTS: No difference in bubble loads was found between the groups at any time despite evident muscle injury. Maximum bubble loads (bubbles cm-2 heart cycle-1) were not different, exercise: 1.6 ± 3.5 SD vs control: 2.2 ± 4.1 SD, P = 0.90, n = 15 in each group. CONCLUSIONS: Eccentric exercise performed 48 h prior to diving causes skeletal muscle injury but does not increase the amount of vascular bubbles in rats. The prevailing recommendation is that physical activity prior to diving is a risk factor of DCS. However, present and previous studies implicate that pre-dive physical activity does not increase the DCS risk

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

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 anaesthetized, 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 one hour after decompression until sacrificed, and showed no gas bubbles during dissection

Fast hyperbaric decompression after heliox saturation altered the brain proteome in rats

Better understanding of the physiological mechanisms and neurological symptoms involved in the development of decompression sickness could contribute to improvements of diving procedures. The main objective of the present study was to determine effects on the brain proteome of fast decompression (1 bar/20 s) compared to controls (1 bar/10 min) after heliox saturation diving, using rats in a model system. The protein S100B, considered a biomarker for brain injury, was not significantly different in serum samples from one week before, immediately after, and one week after the dive. Alterations in the rat brain proteome due to fast decompression were investigated using both iontrap and orbitrap LC-MS, and 967 and 1062 proteins were quantified, respectively. Based on the significantly regulated proteins in the iontrap (56) and orbitrap (128) datasets, the networks “synaptic vesicle fusion and recycling in nerve terminals” and “translation initiation” were significantly enriched in a system biological database analysis (Metacore). Ribosomal proteins (RLA2, RS10) and the proteins hippocalcin-like protein 4 and proteasome subunit beta type-7 were significantly upregulated in both datasets. The heat shock protein 105 kDa, Rho-associated protein kinase 2 and Dynamin-1 were significantly downregulated in both datasets. Another main effect of hyperbaric fast decompression in our experiment is inhibition of endocytosis and stimulation of exocytosis of vesicles in the presynaptic nerve terminal. In addition, fast decompression affected several proteins taking parts in these two main mechanisms of synaptic strength, especially alteration in CDK5/calcineurin are associated with a broad range of neurological disorders. In summary, fast decompression after heliox saturation affected the brain proteome in a rat model for diving, potentially disturbing protein homeostasis, e.g. in synaptic vesicles, and destabilizing cytoskeletal components. Data are available via ProteomeXchange with identifier PXD006349

Proteins regulated in the brain of diving rats in both orbitrap and Iontrap analyses.

<p>Proteins regulated in the brain of diving rats in both orbitrap and Iontrap analyses.</p

Proteins only detected in either orbitrap or Iontrap and significantly regulated.

<p>Proteins only detected in either orbitrap or Iontrap and significantly regulated.</p

Network analysis of the >20% significantly regulated proteins.

<p>The proteins significantly regulated (>20% regulation) due to fast decompression (Tables <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0185765#pone.0185765.t002" target="_blank">2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0185765#pone.0185765.t003" target="_blank">3</a>) were analysed for potential protein-protein interactions and GO enrichments using String. Green circles indicate the significantly downregulated proteins in the datasets, all proteins without a green circle were significantly upregulated. Explanation of protein name abbreviations, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0185765#pone.0185765.t003" target="_blank">Table 3</a>.</p