Hydration status during commercial saturation diving measured by bioimpedance and urine specific gravity.

Excessive fluid loss triggered by hyperbaric pressure, water immersion and hot water suits causes saturation divers to be at risk of dehydration. Dehydration is associated with reductions in mental and physical performance, resulting in less effective work and an increased risk of work-related accidents. In this study we examined the hydration status of 11 male divers over 19 days of a commercial saturation diving campaign to a working depth of 74 m, using two non-invasive methods: Bioelectrical impedance analysis (BIA) and urine specific gravity (USG). Measurements were made daily before and after bell runs, and the BIA data was used to calculated total body water (TBW). We found that BIA and USG were weakly negatively correlated, probably reflecting differences in what they measure. TBW was significantly increased after bell runs for all divers, but more so for bellmen than for in-water divers. There were no progressing changes in TBW over the 19-day study period, indicating that the divers' routines were sufficient for maintaining their hydration levels on short and long term.

A Work Environment Under Pressure: Psychosocial Job Demands and Resources Among Saturation Divers.

Saturation divers work and live under high physiological and social demands for weeks on end. Even though physiological research has contributed insights to the work conditions of saturation divers, research on the qualities of the divers' psychosocial work environment is lacking. This study aimed to explore which job demands and resources are viewed as characteristic among saturation divers working within an isolated and confined environment. Based on data from 6 in-depth semi-structured interviews, template analysis was applied to map unique characteristics. By using the theoretical framework of the job demands-resources model, we found that the work environment in saturation diving was characterized by shifting demands and big contrasts, requiring adaptability in each individual diver. One major demand described by the informants was an unpredictable future, somewhat due to the changes in the oil and gas industry. Another important demand was the conflict between family and work/leisure when committing to work for extended periods in isolated environments. The monotony that characterizes the work environment is a challenge that must be managed. High wages, periods of leisure, and a prestigious job provide external motivation, while personal resources such as mental endurance and flexibility, a willingness to learn, and keeping up small personal routines, may benefit the divers' mental health. This is also affected by the quality of team climate-with features such as being sociable and considerate, having a dark sense of humor and having trust in one another.

Using Salivary Biomarkers for Stress Assessment in Offshore Saturation Diving: A Pilot Study.

Health monitoring during offshore saturation diving is complicated due to restricted access to the divers, the desire to keep invasive procedures to a minimum, and limited opportunity for laboratory work onboard dive support vessels (DSV). In this pilot study, we examined whether measuring salivary biomarkrers in samples collected by the divers themselves might be a feasible approach to environmental stress assessment. Nine saturation divers were trained in the passive drool method for saliva collection and proceeded to collect samples at nine time points before, during, and after an offshore commercial saturation diving campaign. Samples collected within the hyperbaric living chambers were decompressed and stored frozen at -20°C onboard the DSV until they were shipped to land for analysis. Passive drool samples were collected without loss and assayed for a selection of salivary biomarkers: secretory immunoglobulin A (SIgA), C-reactive protein (CRP), tumor necrosis factor (TNF)-α, interleukins IL-6, IL-8, IL-1ß, as well as cortisol and alpha-amylase. During the bottom phase of the hyperbaric saturation, SIgA, CRP, TNF-α, IL-8 and IL-1ß increased significantly, whereas IL-6, cortisol and alpha-amylase were unchanged. All markers returned to pre-dive levels after the divers were decompressed back to surface pressure. We conclude that salivary biomarker analysis may be a feasible approach to stress assessment in offshore saturation diving. The results of our pilot test are consonant with an activation of the sympathetic nervous system related to systemic inflammation during hyperbaric and hyperoxic saturation.

Functional Profiling Reveals Altered Metabolic Activity in Divers' Oral Microbiota During Commercial Heliox Saturation Diving.

Background: The extreme environment in saturation diving affects all life forms, including the bacteria that reside on human skin and mucosa. The oral cavity alone is home to hundreds of different bacteria. In this study, we examined the metabolic activity of oral bacteria from healthy males during commercial heliox saturation diving. We focused on environmentally induced changes that might affect the divers' health and fitness. Methods: We performed pathway abundance analysis using PICRUSt2, a bioinformatics software package that uses marker gene data to compute the metabolic activity of microbial communities. The analysis is based on 16S rRNA metagenomic data generated from the oral microbiota of 23 male divers before, during, and after 4weeks of commercial heliox saturation diving. Environmentally induced changes in bacterial metabolism were computed from differences in predicted pathway abundances at baseline before, versus during, and immediately after saturation diving. Results and Conclusion: The analysis predicted transient changes that were primarily associated with the survival and growth of bacteria in oxygenated environments. There was a relative increase in the abundance of aerobic metabolic pathways and a concomitant decrease in anaerobic metabolic pathways, primarily comprising of energy metabolism, oxidative stress responses, and adenosylcobalamin biosynthesis. Adenosylcobalamin is a bioactive form of vitamin B12 (vitB12), and a reduction in vitB12 biosynthesis may hypothetically affect the divers' physiology. While host effects of oral bacterial vitamin metabolism are uncertain, this is a finding that concurs with the existing recommendations for vitB12 supplements as part of the divers' diet, whether to boost antioxidant defenses in bacteria or their host or to improve oxygen transport during saturation diving.

Effects of Cold Decompression on Hemodynamic Function and Decompression Sickness Risk in a Dry Diving Rat Model.

Background: Diving in cold water is thought to increase the risk of decompression sickness (DCS), especially if the diver is cold during decompression. In this study, we investigated hemodynamic function and DCS risk in an animal model, where cold decompression was followed by rewarming at the surface. Methods: Nine female Sprague Dawley rats had pressure-volume catheters inserted into their left heart ventricle and femoral artery before they were exposed to dry air dives in which their core temperature was normothermic during the bottom phase, cold (35°C) during decompression, and normothermic after the dive. Data from an earlier study were used as controls. The rats were compressed in air to 600kPa, maintained at pressure for 45min, and decompressed at 50kPa/min. Hemodynamic data were recorded before, during, and 60min after decompression. Venous gas bubbles were recorded in the right heart ventricle and pulmonary artery for 60min after the dive. Results and Conclusion: During decompression, cardiac output (CO), and stroke volume (SV) decreased equally in cold rats and controls. CO and SV were temporarily re-established at the surface, before falling again in the cold rats. There was no difference in post-dive venous bubble grades. However, as the post-dive fall in CO and SV could be a sign of gas emboli obstructing the pulmonary circulation, we cannot conclude whether the DCS risk was increased. More sensitive bubble detection methods are needed to elucidate this point.

The Assessment of Daily Energy Expenditure of Commercial Saturation Divers Using Doubly Labelled Water.

Commercial saturation divers are exposed to unique environmental conditions and are required to conduct work activity underwater. Consequently, divers' physiological status is shown to be perturbed and therefore, appropriate strategies and guidance are required to manage the stress and adaptive response. This study aimed to evaluate the daily energy expenditure (DEE) of commercial saturation divers during a 21-day diving operation in the North Sea. Ten saturation divers were recruited during a diving operation with a living depth of 72 metres seawater (msw) and a maximum working dive depth of 81 msw. Doubly labelled water (DLW) was used to calculate DEE during a 10-day measurement period. Energy intake was also recorded during this period by maintaining a dietary log. The mean DEE calculated was 3030.9 ± 513.0 kcal/day, which was significantly greater than the mean energy intake (1875.3 ± 487.4 kcal; p = 0.005). There was also a strong positive correction correlation between DEE and total time spent performing underwater work (r = 0.7, p = 0.026). The results suggested saturation divers were in a negative energy balance during the measurement period with an intraindividual variability in the energy cost present that may be influenced by time spent underwater.

Acute Effects on the Human Peripheral Blood Transcriptome of Decompression Sickness Secondary to Scuba Diving.

Decompression sickness (DCS) develops due to inert gas bubble formation in bodily tissues and in the circulation, leading to a wide range of potentially serious clinical manifestations. Its pathophysiology remains incompletely understood. In this study, we aim to explore changes in the human leukocyte transcriptome in divers with DCS compared to closely matched unaffected controls after uneventful diving. Cases (n = 7) were divers developing the typical cutis marmorata rash after diving with a confirmed clinical diagnosis of DCS. Controls (n = 6) were healthy divers who surfaced from a ≥25 msw dive without decompression violation or evidence of DCS. Blood was sampled at two separate time points-within 8 h of dive completion and 40-44 h later. Transcriptome analysis by RNA-Sequencing followed by bioinformatic analysis was carried out to identify differentially expressed genes and relate their function to biological pathways. In DCS cases, we identified enrichment of transcripts involved in acute inflammation, activation of innate immunity and free radical scavenging pathways, with specific upregulation of transcripts related to neutrophil function and degranulation. DCS-induced transcriptomic events were reversed at the second time point following exposure to hyperbaric oxygen. The observed changes are consistent with findings from animal models of DCS and highlight a continuum between the responses elicited by uneventful diving and diving complicated by DCS. This study sheds light on the inflammatory pathophysiology of DCS and the associated immune response. Such data may potentially be valuable in the search for novel treatments targeting this disease.

Shifts in the Oral Microbiota During a Four-Week Commercial Saturation Dive to 200 Meters.

During commercial saturation diving, divers live and work under hyperbaric and hyperoxic conditions. The myriads of bacteria that live in and on the human body must adjust to the resultant hyperbaric stress. In this study, we examined the shifts in bacterial content in the oral cavity of saturation divers, using a metagenomic approach to determine the diversity in the composition of bacterial phyla and genera in saliva from 23 male divers before, during, and immediately after 4 weeks of commercial heliox saturation diving to a working depth of circa 200 m. We found that the bacterial diversity fell during saturation, and there was a change in bacterial composition; with a decrease at the phylum level of obligate anaerobe Fusobacteria, and an increase of the relative abundance of Actinobacteria and Proteobacteria. At the genus level, Fusobacterium, Leptotrichia, Oribacterium, and Veillonella decreased, whereas Neisseria and Rothia increased. However, at the end of the decompression, both the diversity and composition of the microbiota returned to pre-dive values. The results indicate that the hyperoxic conditions during saturation may suppress the activity of anaerobes, leaving a niche for other bacteria to fill. The transient nature of the change could imply that hyperbaric heliox saturation has no lasting effect on the oral microbiota, but it is unknown whether or how a shift in oral bacterial diversity and abundance during saturation might impact the divers' health or well-being.

Endothelial dysfunction in small arteries and early signs of atherosclerosis in ApoE knockout rats.

Endothelial dysfunction is recognized as a major contributor to atherosclerosis and has been suggested to be evident far before plaque formation. Endothelial dysfunction in small resistance arteries has been suggested to initiate long before changes in conduit arteries. In this study, we address early changes in endothelial function of atherosclerosis prone rats. Male ApoE knockout (KO) rats (11- to 13-weeks-old) were subjected to either a Western or standard diet. The diet intervention continued for a period of 20-24 weeks. Endothelial function of pulmonary and mesenteric arteries was examined in vitro using an isometric myograph. We found that Western diet decreased the contribution of cyclooxygenase (COX) to control the vascular tone of both pulmonary and mesenteric arteries. These changes were associated with early stage atherosclerosis and elevated level of plasma total cholesterol, LDL and triglyceride in ApoE KO rats. Chondroid-transformed smooth muscle cells, calcifications, macrophages accumulation and foam cells were also observed in the aortic arch from ApoE KO rats fed Western diet. The ApoE KO rats are a new model to study endothelial dysfunction during the earlier stages of atherosclerosis and could help us improve preclinical drug development.

Extensive Simulated Diving Aggravates Endothelial Dysfunction in Male Pro-atherosclerotic ApoE Knockout Rats.

INTRODUCTION: The average age of the diving population is rising, and the risk of atherosclerosis and cardiovascular disease in divers are accordingly increasing. It is an open question whether this risk is altered by diving per se. In this study, we examined the effect of 7-weeks simulated diving on endothelial function and mitochondrial respiration in atherosclerosis-prone rats. METHODS: Twenty-four male ApoE knockout (KO) rats (9-weeks-old) were fed a Western diet for 8 weeks before 12 rats were exposed to simulated heliox dry-diving in a pressure chamber (600 kPa for 60 min, decompression of 50 kPa/min). The rats were dived twice-weekly for 7 weeks, resulting in a total of 14 dives. The remaining 12 non-diving rats served as controls. Endothelial function of the pulmonary and mesenteric arteries was examined in vitro using an isometric myograph. Mitochondrial respiration in cardiac muscle tissues was measured using high-resolution respirometry. RESULTS AND CONCLUSION: Both ApoE KO diving and non-diving rats showed changes in endothelial function at the end of the intervention, but the extent of these changes was larger in the diving group. Altered nitric oxide signaling was primarily involved in these changes. Mitochondrial respiration was unaltered. In this pro-atherosclerotic rat model of cardiovascular changes, extensive diving appeared to aggravate endothelial dysfunction rather than promote adaptation to oxidative stress.

Hemoglobin During and Following a 4-Week Commercial Saturation Dive to 200 m.

Commercial saturation divers must acclimatize to hyperbaric hyperoxia in their work environment, and subsequently readjust to breathing normal air when their period in saturation is over. In this study, we measured hemoglobin (Hb) during and following 4 weeks of heliox saturation diving in order to monitor anemia development and the time for Hb to recover post-saturation. Male commercial saturation divers reported their capillary blood Hb daily, before, and during 28 days of heliox saturation to a working depth of circa 200 m (n = 11), and for 12 days at surface post-saturation (n = 9-7), using HemoCue 201+ Hb devices. Hb remained in normal range during the bottom phase, but fell during the decompression; reaching levels of mild anemia (≤13.6 g/dl) the day after the divers' return to the surface. Hb was significantly lower than the pre-saturation baseline (14.7 ± 1.1 g/dl) on the fifth day post-saturation (12.8 ± 1.8 g/dl, p = 0.028), before reverting to normal after 6-7 days. At the end of the 12-day post-saturation period, Hb was not statistically different from the pre-saturation baseline. The observed Hb changes, although significant, were modest. While we cannot rule out effect of other factors, the presence of mild anemia may partially explain the transient fatigue that commercial saturation divers experience post-saturation.

A Single Simulated Heliox Dive Modifies Endothelial Function in the Vascular Wall of ApoE Knockout Male Rats More Than Females.

INTRODUCTION: The number of divers is rising every year, including an increasing number of aging persons with impaired endothelial function and concomitant atherosclerosis. While diving is an independent modulator of endothelial function, little is known about how diving affects already impaired endothelium. In this study, we questioned whether diving exposure leads to further damage of an already impaired endothelium. METHODS: A total of 5 male and 5 female ApoE knockout (KO) rats were exposed to simulated diving to an absolute pressure of 600 kPa in heliox gas (80% helium, 20% oxygen) for 1 h in a dry pressure chamber. 10 ApoE KO rats (5 males, 5 females) and 8 male Sprague-Dawley rats served as controls. Endothelial function was examined in vitro by isometric myography of pulmonary and mesenteric arteries. Lipid peroxidation in blood plasma, heart and lung tissue was used as measures of oxidative stress. Expression and phosphorylation of endothelial NO synthase were quantified by Western blot. RESULTS AND CONCLUSION: A single simulated dive was found to induce endothelial dysfunction in the pulmonary arteries of ApoE KO rats, and this was more profound in male than female rats. Endothelial dysfunction in males was associated with changing in production or bioavailability of NO; while in female pulmonary arteries an imbalance in prostanoid signaling was observed. No effect of diving was found on mesenteric arteries from rats of either sex. Our findings suggest that changes in endothelial dysfunction were specific for pulmonary circulation. In future, human translation of these findings may suggest caution for divers who are elderly or have prior reduced endothelial function.

Continuous Hemodynamic Monitoring in an Intact Rat Model of Simulated Diving.

Cardiovascular risk is elevated in divers, but detailed information of cardiac function during diving is missing. The aim of this study was to apply an intact rat model with continuous monitoring of cardiac left ventricular (LV) function in a simulated diving experiment. Thirteen rats were inserted with a LV pressure-volume catheter and a pressure transducer in the femoral artery to measure hemodynamic variables, and randomly assigned to diving (n = 9) and control (n = 4) groups. The diving group was compressed to 600 kPa in air, maintained at pressure for 45 min (bottom phase), and decompressed to surface at 50 kPa/min. Data was collected before, during, and up to 60 min after exposure in the diving group, and at similar times in non-diving controls. During the bottom phase, stroke volume (SV) (-29%) and cardiac output (-30%) decreased, whereas LV end-systolic volume (+13%), mean arterial pressure (MAP) (+29%), and total peripheral resistance (TPR) (+72%) increased. There were no changes in LV contractility, stroke work, or diastolic function. All hemodynamic variables returned to baseline values within 60 min after diving. In conclusion, our simulated dive experiment to 600 kPa increased MAP and TPR to levels which caused a substantial reduction in SV and LV volume output. The increase in cardiac afterload demonstrated to take place during a dive is well tolerated by the healthy heart in our model, whereas in a failing heart this abrupt change in afterload may lead to acute cardiac decompensation.

Hemoglobin and Erythropoietin After Commercial Saturation Diving.

Saturation divers are exposed to elevated partial pressure of oxygen (ppO2) in their hyperbaric work environment. Experimental studies indicate that oxygen transport is altered, and we have previously reported a drop in hematocrit and extensive downregulation of genes involved in blood oxygen transport capacity after decompression from professional saturation diving. Here we investigate the initial period of hematological adjustment back to normobaric air after professional saturation diving. Erythropoietin (EPO) and hemoglobin (Hb) were measured in blood from 13 divers at two time-points after saturation assignments lasting up to 4 weeks; first immediately after decompression and again 24 h later. Pre-dive levels defined baselines. The ppO2 varied from 40 kPa in the saturation chambers during storage, 50 to 80 kPa during bell excursions, and gradually reduced to 21 kPa during decompression to surface pressure. EPO was similar to baseline immediately after saturation diving (P = 0.4), and markedly increased within the next 24 h (99%, P < 0.0002). Hb levels remained slightly reduced at both time-points (4% immediately after; P = 0.02, 8% 24 h after; P < 0.001). The results imply that the hematological acclimatization back to normobaric air was ongoing, but not completed, during the first 24 h after professional saturation diving.

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.

Commercial Divers' Subjective Evaluation of Saturation.

Commercial saturation diving involves divers living and working in an enclosed atmosphere with elevated partial pressure of oxygen (ppO2) for weeks. The divers must acclimatize to these conditions during compression, and for up to 28 days until decompression is completed. During decompression, the ppO2 and ambient pressure are gradually decreased; then the divers must acclimatize again to breathing normal air in atmospheric pressure when they arrive at surface. We investigated 51 saturation divers' subjective evaluation of the saturation and post-decompression phase via questionnaires and individual interviews. The questions were about decompression headaches and fatigue; and time before recovering to a pre-saturation state. Twenty-two (44%) of the divers who responded declared having headaches; near surface (44%) or after surfacing (56%). 71% reported post-saturation fatigue after their last saturation, 82% of them described it as typical and systematic after each saturation. Recovery was reported to normally take from 1 to 10 days. The fatigue and headaches observed are compatible with divers' acclimatization to the changes in ppO2 levels during saturation and decompression. They appear to be reversible post- decompression.