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.

MRI of the central nervous system in rats following heliox saturation decompression.

AIMS: The main objectives of the present study was to establish an animal model of decompression sickness (DCS) after heliox saturation diving, and to use this model to evaluate possible morphological changes in the CNS induced by DCS using structural MRI. METHODS: Two groups of rats were pressurized with heliox to 5 bar (pO2 = 50 kPa). The saturation time was three hours; decompression rate was 1 bar/10 seconds or 1 bar/20 seconds. A 7.0 Tesla small animal MRI scanner was used for detection of possible morphological changes in the brain and spinal cord, two hours and one week after the dive, compared to one week prior to the dive. RESULTS: Neurological symptoms of DCS were observed in seven out of 10 animals. MRI of the brain and spinal cord did not reveal any morphological CNS injuries. CONCLUSION: This diving procedure was successful in causing DCS in a large proportion of the animals. However, despite massive neurological signs of DCS, no visible CNS injuries were observed in the MRI scans.

Natural killer cells as biomarkers of hyperbaric stress during a dry heliox saturation dive.

INTRODUCTION: Diving, hyperbaric oxygen, and decompression have been described as inducers of alterations in various components of the human immune system, such as the distribution of circulating lymphocytes. Hypothetically, the monitoring of specific lymphocyte subsets during hyperbaric exposure, including T- and NK-cell subsets, can serve as biomarkers of hyperbaric stress. METHODS: Eight experienced saturation divers and eight reference subjects, naive to deep saturation diving, were examined. Peripheral blood mononuclear cells were isolated before and at different points during a 19.3-d dry heliox saturation dive to 2.64 MPa (254 msw). The NK cell cytotoxicity was estimated in a 4-h 51Cr-release assay using the NK cell sensitive tumor cell-line K562 as target cells. The major lymphocyte subpopulations, with special emphasis on the NK cell subsets, were phenotypically delineated by the use of 4-color flow cytometry. RESULTS: Although NK cell cytotoxicity increased significantly in the divers during the compression phase and the reference subjects who remained in normoxic conditions outside the chamber, the NK cell cytotoxicity was significantly higher in the divers. DISCUSSION: This finding, together with augmentation in the absolute number of circulating NK cells in the divers due to a possible activation of specific parts of the innate cellular immune system during hyperbaric exposure, suggests the monitoring of specific immune functions can be useful as biomarkers of hyperbaric-induced inflammatory stress.

A subjective evaluation of a drinking system for saturation divers.

Studies have shown that divers may lose large volumes of body fluids in hot water suit (HWS) dives lasting for four hours or longer, and that this dehydration is mainly caused by sweating. Body fluid balance may be impaired and the diver's alertness and power of judgement could be influenced by such imbalance. The main objective of the present study was to obtain a subjective judgement of a drinking system for divers (DSFD) and to obtain information related to body fluid loss during long saturation lock-out dives. Via a suction pipe imbedded in the microphone unit in the oronasal mask, the DSFD makes it possible for the diver to drink while in the water. Ten divers tested the drinking system during 12 saturation lock-out dives lasting on average for 5.5 h. A questionnaire was answered after each dive. The divers drank 21 times (range 5-30 times) during the dives, and the average drinking volume was 1.4 litre (range 1.0-1.5 litre) but only drank 0.04 litre (range 0-0.3 litre) in the bell after diving. The system was easy to operate and preparation and clothing did not cause any delay. The suction pipe did not intrude and the microphone performed excellently. The work in water was not hindered by DSFD and all divers were very satisfied with the drinking system. It was obvious that the need for fluid intake after a dive with DSFD was markedly reduced; another good indication of maintained body fluid balance.

Postexercise reduction in lung diffusion capacity is not attenuated by skin cooling.

Pulmonary diffusion capacity for carbon monoxide (DL(CO)) is reduced by approximately 10% 1-6 h after maximal exercise. The mechanisms may be interstitial alveolar oedema and reduced pulmonary capillary blood volume. It was hypothesized that thermal stress following exercise contributes to the reduction in DL(CO), and that skin cooling would attenuate the postexercise reduction in DL(CO). Cutaneous vascular conductance (CVC), mean surface temperature (MST), rectal temperature and DL(CO) were measured before and 90 min after maximal incremental cycle exercise. Thereafter, the subjects were exposed to cold air without eliciting shivering one day and another day served as control. The measurements were repeated 120 min after exercise. Twelve healthy subjects (six male) aged 20-27 years were studied. DL(CO) was reduced by 7.1% (SD = 6.3%, P = 0.003) and 7.6% (SD = 5.3%, P<0.001) 90 and 120 min after exercise in the control experiment. It was reduced by 5.6% (SD = 5.5%, P = 0.014) 90 min after exercise and remained reduced by 6.1% (SD = 6.1%, P = 0.012) after cooling despite a significant reduction in CVC and in MST from 31.9 (SD = 0.6) degrees C to 27.4 (SD = 1.9) degrees C. We conclude that the postexercise reduction in DL(CO) is present when thermal status is restored after exercise, and that it is not influenced by further skin surface cooling.

Venous gas emboli in normal and dehydrated rats following decompression from a saturation dive.

INTRODUCTION: Dehydration may increase the risk for decompression sickness (DCS). Since DCS most probably is caused by endogenous gas phase formation, we hypothesized that decompression will induce more venous gas emboli (VGE) in dehydrated rats compared to controls. METHODS: Two groups of rats were pressurized to 0.5 MPa (5 ATA) on heliox for 16 h, and thereafter decompressed to atmospheric pressure at a rate of 0.3 MPa x min(-1). The nine control rats had free access to water ad libitum whereas the eight dehydrated rats were water-deprived for 48 h before decompression. During and after decompression, VGE was measured in the vena cava for 60 min with the Doppler technique and graded into six bubble grade (BG) categories. Body mass (BM), and food and water intake were registered daily, and venous blood samples were taken before and after pressure exposure. RESULTS: Serum osmolality and hematocrit increased significantly in dehydrated rats (306 +/- 5.2 to 315 +/- 7.3 mosmol x kg(-1) and 39.3 +/- 4.9 to 49.6 +/- 5.2%) but not in controls (300 +/- 8.9 to 303 +/- 6.7 mosmol x kg(-1) and 40.3 +/- 5.2 to 41.4 +/- 6.1%). Plasma volume decreased by 9.2% (P < 0.05) and 2.8% (n.s.) in dehydrated and control rats. VGE were detected in all control animals (average BG: 2.8 +/- 1.9), but only in four water-deprived rats (BG: 1.6 +/- 2.2). This difference was not significant. CONCLUSIONS: Our experiments do not support the idea that dehydration increases circulatory VGE.

Time and temperature effects on body fluid loss during dives with the open hot-water suit.

BACKGROUND: Bodyweight (BW) losses up to 5 kg have been observed during diving with the open hot-water suit (HWS). The objective of these dives was to study the hormonal, hematological, and renal effects of dehydration during shallow HWS diving. METHODS: In series 1, four divers dove for 3.5 h each day for 7 d. In series 2, 12 divers dove to 6-8 msw for 1, 2, and 4 h. Blood and urine samples, BW measurements, oral temperature, and thermal stress indices were collected. RESULTS: Average deltaBW (+/- SD) for the 28 dives in series 1 was 1.5 +/- 0.8 kg, and the largest BW reductions were 3.2 and 3.0 kg, corresponding to 3.7 and 4.7% of BW. Changes in thermal stress, hemoglobin, hematocrit, aldosterone, and electrolyte excretion correlated with BW reduction. In series 2, average BW reductions were 0.46 +/- 0.27, 0.96 +/- 0.38, and 1.55 +/- 0.59 kg during 1-, 2-, and 4-h dives. BW reduction correlated significantly with thermal stress (p < 0.01). Aldosterone increased after 1 and 2 h and plasma renin activity was unchanged. Atrial natriuretic peptide increased in all dives (p < 0.01) and arginine vasopressin increased in the 4-h dives (p < 0.05). The 7.2% decrease in plasma volume, the increases in hemoglobin, hematocrit and serum proteins, and an unchanged plasma osmolality indicate an isotonic dehydration after the 4-h dives. CONCLUSIONS: BW loss during HWS diving is mainly caused by sweating. Dives of 4 h produce an isotonic dehydration and a break for fluid intake is, therefore, recommended.

Changes in erythropoietin and haemoglobin concentrations in response to saturation diving.

A reduction in haemoglobin concentration is consistently reported after deep saturation dives. This may be due to a downregulation of erythropoietin (EPO) concentration or to a toxic effect of the hyperoxia associated with the dives resulting in an increased destruction rate of erythrocytes. In this study haemoglobin concentration, blood cell counts, serum ferritin, bilirubin, haptoglobin and EPO concentrations were measured before, during and after a 19 day saturation dive to 240 m. The partial pressure of oxygen (PO(2)) was 35-70 kPa during the 7 day compression and bottom phase, and 30-50 kPa during the 12 day decompression phase. There was a reduction in EPO concentration from 8.4+/-1.4 (mean +/- 1SD) to 6.3 +/- 1.9 U.L(-1) on Dive day 2. On Dive days 7 and 17 EPO concentrations were not significantly different from baseline despite the continued exposure to hyperoxia. Immediately after the dive and return to a normoxic environment there was an increase in the EPO concencentration to 14.5 +/- 4.7 U.L(-1). Haemoglobin concentration, erythrocyte and reticulocyte counts were decreased at the end of the dive, and there was an increase in serum ferritin. There were no changes in bilirubin or haptoglobin concentrations indicative of haemolysis. It appears that the change in PO(2), rather than the sustained exposure to a hyperoxic environment, induces the changes in the EPO concentrations and erythropoietic activity.

Gas bubbles in rats after heliox saturation and different decompression steps and rates.

Effects of pressure reduction, decompression rate, and repeated exposure on venous gas bubble formation were determined in five groups (GI, GII, GIII, GIV, and GV) of conscious and freely moving rats in a heliox atmosphere. Bubbles were recorded with a Doppler ultrasound probe implanted around the inferior caval vein. Rats were held for 16 h at 0.4 MPa (GI), 0.5 MPa (GII and GIII), 1.7 MPa (GIVa), or 1.9 MPa (GIV and GV), followed by decompression to 0.1 MPa in GI to GIII and to 1.1 MPa in GIV and GV. A greater decompression step, but at the same rate (GII vs. GI and GIVb vs. GIVa), resulted in significantly more bubbles (P < 0.01). A twofold decompression step resulted in equal amount of bubbles when decompressing to 1.1 MPa compared with 0.1 MPa. The faster decompression in GII and GVa (10.0 kPa/s) resulted in significantly more bubbles (P < 0.01) compared with GIII and GVb (2.2 kPa/s). No significant difference was observed in cumulative bubble score when comparing first and second exposure. With the present animal model, different decompression regimes may be evaluated.