Elevations of Extracellular Vesicles and Inflammatory Biomarkers in Closed Circuit SCUBA Divers.

Blood-borne extracellular vesicles and inflammatory mediators were evaluated in divers using a closed circuit rebreathing apparatus and custom-mixed gases to diminish some diving risks. "Deep" divers (n = 8) dove once to mean (±SD) 102.5 ± 1.2 m of sea water (msw) for 167.3 ± 11.5 min. "Shallow" divers (n = 6) dove 3 times on day 1, and then repetitively over 7 days to 16.4 ± 3.7 msw, for 49.9 ± 11.9 min. There were statistically significant elevations of microparticles (MPs) in deep divers (day 1) and shallow divers at day 7 that expressed proteins specific to microglia, neutrophils, platelets, and endothelial cells, as well as thrombospondin (TSP)-1 and filamentous (F-) actin. Intra-MP IL-1ß increased by 7.5-fold (p < 0.001) after day 1 and 41-fold (p = 0.003) at day 7. Intra-MP nitric oxide synthase-2 (NOS2) increased 17-fold (p < 0.001) after day 1 and 19-fold (p = 0.002) at day 7. Plasma gelsolin (pGSN) levels decreased by 73% (p < 0.001) in deep divers (day 1) and 37% in shallow divers by day 7. Plasma samples containing exosomes and other lipophilic particles increased from 186% to 490% among the divers but contained no IL-1ß or NOS2. We conclude that diving triggers inflammatory events, even when controlling for hyperoxia, and many are not proportional to the depth of diving.

Pulmonary Effects of One Week of Repeated Recreational Closed-Circuit Rebreather Dives in Cold Water.

Background and Objectives: The use of closed-circuit rebreathers (CCRs) in recreational diving is gaining interest. However, data regarding its physiological effects are still scarce. Immersion, cold water, hyperoxia, exercise or the equipment itself could challenge the cardiopulmonary system. The purpose of this study was to examine the impact of CCR diving on lung function and autonomous cardiac activity after a series of CCR dives in cold water. Materials and Methods: Eight CCR divers performed a diving trip (one week) in the Baltic Sea. Spirometry parameters, SpO2, and the lung ultrasonography score (LUS) associated with hydration monitoring by bioelectrical impedance were assessed at the end of the week. Heart rate variability (HRV) was recorded during the dives. Results: No diver declared pulmonary symptoms. The LUS increased after dives combined with a slight non-pathological decrease in SpO2. Spirometry was not altered, and all body water compartments were increased. Global HRV decreased during diving with a predominant increase in sympathetic tone while the parasympathetic tone decreased. All parameters returned to baseline 24 h after the last dive. Conclusions: The lung aeration disorders observed seem to be transient and not associated with functional spirometry alteration. The HRV dynamics highlighted physiological constraints during the dive as well as environmental-stress-related stimulation that may influence pulmonary changes. The impact of these impairments is unknown but should be taken into account, especially when considering long and repetitive CCR dives.

Physiological effects of mixed-gas deep sea dives using a closed-circuit rebreather: a field pilot study.

PURPOSE: Deep diving using mixed gas with closed-circuit rebreathers (CCRs) is increasingly common. However, data regarding the effects of these dives are still scarce. This preliminary field study aimed at evaluating the acute effects of deep (90-120 msw) mixed-gas CCR bounce dives on lung function in relation with other physiological parameters. METHODS: Seven divers performed a total of sixteen open-sea CCR dives breathing gas mixture of helium, nitrogen and oxygen (trimix) within four days at 2 depths (90 and 120 msw). Spirometric parameters, SpO2, body mass, hematocrit, short term heart rate variability (HRV) and critical flicker fusion frequency (CFFF) were measured at rest 60 min before the dive and 120 min after surfacing. RESULTS: The median [1st-3rd quartile] of the forced vital capacity was lower (84% [76-93] vs 91% [74-107] of predicted values; p = 0.029), whereas FEV1/FVC was higher (98% [95-99] vs 95% [89-99]; p = 0.019) after than before the dives. The other spirometry values and SpO2 were unchanged. Body mass decreased from 73.5 kg (72.0-89.6) before the dives to 70.0 kg (69.2-85.8) after surfacing (p = 0.001), with no change of hematocrit or CFFT. HRV was increased as indicated by the higher SDNN, RMSSD and pNN50 after than before dives. CONCLUSION: The present observation represents the first original data regarding the effects of deep repeated CCR dives. The body mass loss and decrease of FVC after bounce dives at depth of about 100 msw may possibly impose an important physiological stress for the divers.

Gene expression of Decompression Sickness-resistant rats through a miRnome/transcriptome crossed approach.

Susceptibility to decompression sickness (DCS) is characterized by a wide inter-individual variability, the origins of which are still poorly understood. We selectively bred rats with at least a 3-fold greater resistance to DCS than standard rats after 6 generations. In order to better understand DCS mechanisms, we compared the static genome expression of these resistant rats from the 10th generation to their counterparts of the initial non-resistant Wistar strain, by a microarray transcriptomic approach coupled and crossed with a PCR plates miRnome study. Thus, we identified differentially expressed genes on selected males and females, as well as gender differences in those genes, and we crossed these transcripts with the respective targets of the differentially expressed microRNAs. Our results highlight pathways involved in inflammatory responses, circadian clock, cell signaling and motricity, phagocytosis or apoptosis, and they confirm the importance of inflammation in DCS pathophysiology.

A review of nutritional recommendations for scuba divers.

BACKGROUND: Scuba diving is an increasingly popular activity that involves the use of specialized equipment and compressed air to breathe underwater. Scuba divers are subject to the physiological consequences of being immersed in a high-pressure environment, including, but not limited to, increased work of breathing and kinetic energy expenditure, decreased fluid absorption, and alteration of metabolism. Individual response to these environmental stressors may result in a differential risk of decompression sickness, a condition thought to result from excess nitrogen bubbles forming in a diver's tissues. While the mechanisms of decompression sickness are still largely unknown, it has been postulated that this response may further be influenced by the diver's health status. Nutritional intake has direct relevancy to inflammation status and oxidative stress resistance, both of which have been associated with increased decompression stress. While nutritional recommendations have been determined for saturation divers, these recommendations are likely overly robust for recreational divers, considering that the differences in time spent under pressure and the maximum depth could result nonequivalent energetic demands. Specific recommendations for recreational divers remain largely undefined. METHODS: This narrative review will summarize existing nutritional recommendations and their justification for recreational divers, as well as identify gaps in research regarding connections between nutritional intake and the health and safety of divers. RESULTS: Following recommendations made by the Institute of Medicine and the Naval Medical Research Institute of Bethesda, recreational divers are advised to consume ~170-210 kJ·kg-1 (40-50 kcal·kg-1) body mass, depending on their workload underwater, in a day consisting of 3 hours' worth of diving above 46 msw. Recommendations for macronutrient distribution for divers are to derive 50% of joules from carbohydrates and less than 30% of joules from fat. Protein consumption is recommended to reach a minimum of 1 g of protein·kg-1 of body mass a day to mitigate loss of appetite while meeting energetic requirements. All divers should take special care to hydrate themselves with an absolute minimum of 500 ml of fluid per hour for any dive longer than 3 hours, with more recent studies finding 0.69 liters of water two hours prior to diving is most effective to minimize bubble loads. While there is evidence that specialized diets may have specific applications in commercial or military diving, they are not advisable for the general recreational diving population considering the often extreme nature of these diets, and the lack of research on their effectiveness on a recreational diving population. CONCLUSIONS: Established recommendations do not account for changes in temperature, scuba equipment, depth, dive time, work of breathing, breathing gas mix, or individual variation in metabolism. Individual recommendations may be more accurate when accounting for basal metabolic rate and physical activity outside of diving. However, more research is needed to validate these estimates against variation in dive profile and diver demographics.

Scientific shallow saturation dive expedition using diving rebreathers and a specific dry habitat: medical management of the "Capsule" programme.

BACKGROUND: Scientific underwater exploration could benefit from professional diving facilities. This could allow marine research for durations far exceeding anything currently possible. The closed-circuit rebreather expansion provides new perspectives by unleashing divers and their diving bell. "Under the Pole Expeditions" developed an innovative compact underwater habitat for this purpose. MATERIALS AND METHODS: The habitat's depth was fixed at 20 m. Saturation lasted 3 days and was followed by a 245 min long decompression procedure with mandatory in-water phase. Isolation and environmental constraints will require specific medical and safety procedures. "In situ" medical concerns were considered, and a specific evacuation plan was established. This report describes the medical management of this atypical project and the systematic clinical follow-up mostly targeted on the cardiovascular system, fatigue and psychological tolerance. RESULTS: Seventeen individual saturation exposures were performed. All selected divers were professional. Neither severe illness nor decompression sickness was observed. These short-term saturation exposures appeared to be well tolerated. There was a relatively low bubble grade after decompression. Psychological tolerance appeared good. However, a transient moderate orthostatic hypotension suggested cardiovascular deconditioning after dive. CONCLUSIONS: This first experiment demonstrates the interest and feasibility of a shallow revisited saturation dive with rebreather use. This isolation requires medical accompaniment and rigorous preparation. Medical and physiological risks assessment is essential in this context and must be consolidated by new experiences.

Highlighting of the interactions of MYD88 and NFKB1 SNPs in rats resistant to decompression sickness: toward an autoimmune response.

Decompression sickness (DCS) with neurological disorders includes an inappropriate inflammatory response which degenerates slowly, even after the disappearance of the bubbles. There is high inter-individual variability in terms of the occurrence of DCS that could have been mastered by the selection and then the breeding of DCS-resistant rats. We hypothesized the selection of single-nucleotide polymorphisms (SNPs) linked to autoimmunity operated upon a generation of a DCS-resistant strain of rats. We used the candidate gene approach and targeted SNPs linked to the signaling cascade that directly regulates inflammation of innate immunity transiting by the Toll-like receptors. Twenty candidate SNPs were investigated in 36 standard rats and 33 DCS-resistant rats. For the first time, we identify a diplotype (i.e., with matched haplotypes)-when coinherited-that strengthens protection against DCS, which is not strictly homozygous and suggests that a certain tolerance may be considered. We deduced an ideal haplotype of six variants from it (MyD88_50-T, _49-A, _97-C coupled to NFKB_85-T, _69-T, _45-T) linked to the resistant phenotype. Four among the six identified variants are located in pre- and/or post-transcriptional areas regulating MyD88 or NFKB1 expression. Because of missense mutations, the other two variants induce a structural change in the NFKB1 protein complex including one damage alteration according to the Missense3D algorithm. In addition to the MyD88/NFKB1 haplotype providing rats with a strong resistance to DCS, this also highlights the importance that the immune response, here linked to the genetic heritage, can have in the development of DCS and offer a new perspective for therapeutic strategies.

Blood pressure in rats selectively bred for their resistance to decompression sickness.

INTRODUCTION: Susceptibility to decompression sickness (DCS) is characterised by a wide inter-individual variability whose origins are still poorly understood. This hampers reliable prediction of DCS by decompression algorithms. We previously selectively bred rats with a 3-fold greater resistance to DCS than standard rats. Based on its previously reported relation with decompression outcomes, we assessed whether modification in vascular function is associated with resistance to DCS. METHODS: The arterial pressure response to intravenous administration of acetylcholine (ACh, 5 µg.kg-1) and adrenaline (5 and 10 µg.kg-1) was compared in anaesthetised DCS-resistant rats (seven females, seven males) and standard Wistar rats (seven females, 10 males) aged 14-15 weeks. None of these rats had previously undergone hyperbaric exposure. RESULTS: There was a non-significant tendency for a lower diastolic (DBP) and mean blood pressure (MBP) in DCS-resistant rats. After ACh administration, MBP was significantly lower in resistant rats, for both males (P = 0.007) and females (P = 0.034). After administration of adrenaline 10 µg.kg-1, DCS-resistant rats exhibited lower maximal DBP (P = 0.016) and MBP (P = 0.038). Systolic and pulse blood pressure changes did not differ between groups in any of the experiments. CONCLUSIONS: Resistance to DCS in rats is associated to a trend towards a lower vascular tone but not blood pressure reactivity. Whether these differences are a component of the susceptibility to DCS remains to be confirmed.

Evidence of a hormonal reshuffle in the cecal metabolome fingerprint of a strain of rats resistant to decompression sickness.

On one side, decompression sickness (DCS) with neurological disorders lead to a reshuffle of the fecal metabolome from rat caecum. On the other side, there is high inter-individual variability in terms of occurrence of DCS. One could wonder whether the fecal metabolome could be linked to the DCS-susceptibility. We decided to study male and female rats selected for their resistance to decompression sickness, and we hypothesize a strong impregnation concerning the fecal metabolome. The aim is to verify whether the rats resistant to the accident have a fecal metabolomic signature different from the stem generations sensitive to DCS. 39 DCS-resistant animals (21 females and 18 males), aged 14 weeks, were compared to 18 age-matched standard Wistar rats (10 females and 8 males), i.e., the same as those we used for the founding stock. Conventional and ChemRICH approaches helped the metabolomic interpretation of the 226 chemical compounds analyzed in the cecal content. Statistical analysis shows a panel of 81 compounds whose expression had changed following the selection of rats based on their resistance to DCS. 63 compounds are sex related. 39 are in common. This study shows the spectral fingerprint of the fecal metabolome from the caecum of a strain of rats resistant to decompression sickness. This study also confirms a difference linked to sex in the metabolome of non-selected rats, which disappear with selective breeding. Results suggest hormonal and energetic reshuffle, including steroids sugars or antibiotic compounds, whether in the host or in the microbial community.

Physiological characteristics associated with increased resistance to decompression sickness in male and female rats.

Decompression sickness (DCS) is a complex and poorly understood systemic disease with wide interindividual resistance variability. We selectively bred rats with a threefold greater resistance to DCS than standard ones. To investigate possible physiological mechanisms underlying the resistance to DCS, including sex-related differences in these mechanisms, 15 males and 15 females resistant to DCS were compared with aged-matched standard Wistar males (n = 15) and females (n = 15). None of these individuals had been previously exposed to hyperbaric treatment. Comparison of the allelic frequencies of single nucleotide polymorphisms (SNPs) showed a difference of one SNP located on the X chromosome. Compared with nonresistant rats, the neutrophil-to-lymphocyte ratio and the plasmatic activity of coagulation factor X were significantly higher in DCS-resistant individuals regardless of their sex. The maximal relaxation elicited by sodium nitroprusside was lower in DCS-resistant individuals regardless of their sex. Males but not females resistant to DCS exhibited higher neutrophil and lymphocyte counts and higher prothrombin time but lower mitochondrial basal O2 consumption and citrate synthase activity. Principal components analysis showed that two principal components discriminate the DCS-resistant males but not females from the nonresistant ones. These components were loaded with activated partial thromboplastin time, monocyte-to-lymphocyte ratio, prothrombin time, factor X, and fibrinogen for PC1 and red blood cells count and neutrophils count for PC2. In conclusion, the mechanisms that drive the resistance to DCS appear different between males and females; lower coagulation tendency and enhanced inflammatory response to decompression stress might be key for resistance in males. The involvement of these physiological adaptations in resistance to DCS must now be confirmed.NEW & NOTEWORTHY By selective breeding of individuals resistant to decompression sickness (DCS) we previously obtained a rat model of inherited resistance to this pathology. Comparison of these individuals with nonresistant animals revealed differences in leukocyte counts, coagulation, and mitochondrial and vascular functions, but not resistance to oxidative stress. This study also reveals sex-related differences in the physiological changes associated with DCS resistance. A principal components analysis of our data allowed us to discriminate DCS-resistant males from standard ones, but not females. These differences represent possible mechanisms driving resistance to DCS. Although still far from the diver, this opens a pathway to future adaptation of personalized decompression procedures for "DCS-prone" individuals.

A survey of scuba diving-related injuries and outcomes among French recreational divers.

INTRODUCTION: Few studies are available to appreciate the broad spectrum of dive-related injuries (DI), which are not limited to decompression illness (DCI) and fatalities. Studies supporting injury-management efficacy from early recognition to first-aid, final treatment and outcome are also lacking. This study aims at making an epidemiologic inventory of DI among French scuba divers. METHODS: This online, retrospective, cross-sectional survey analyzed self-reported symptoms, context of occurrence, initial response and outcome. The relationships between symptoms and diver characteristics were assessed and severity scores created from the reports. RESULTS: A total of 799 divers responded, of whose questionnaires 784 were sufficiently complete to be analyzed. Approximately one-third (35%) of respondents had never experienced a DI. DCI-like symptoms represent a small fraction of DIs, the most commonly reported being ear barotrauma. Self-reported symptom rates decreased with increasing age and male sex. The ranking dive leader was the primary care provider in 58% of reports and 32% of injured divers never sought help. Management decisions (first aid and/or hyperbaric oxygen treatment) were related to the severity score. Complete resolution was achieved in 84 (74%) of 114 DCI cases, whilst mild (n = 22, 19%) and severe (n = 8, 7%) residual symptoms were reported. One in 10 divers who did not seek treatment for symptoms believed to be related to DCI declared some residual symptoms. CONCLUSION: Based on these results, diving injury rates may be higher than previously reported. However, the most frequent symptoms appear to be of only a modest nature.

A new form of admissible pressure for Haldanian decompression models.

In this article, we propose and study a new form of admissible pressure in the Haldanian framework. We then use it to study the surjectivity of the Gradient Factors on the space of the reachable decompression profiles, and investigate a particular case. This case leads to the proposition of a decompression strategy, whose crucial parameter is the ascent rate. An appropriate ascent rate is suggested as recommended by COMEX, through a physiologically relevantmethod. This new strategy enables the unification of the COMEX approach (not based on a tissuesaturation theory), with the Haldanian method.

Angiotensin Converting Enzyme Inhibitor Has a Protective Effect on Decompression Sickness in Rats.

Introduction: Commercial divers, high altitude pilots, and astronauts are exposed to some inherent risk of decompression sickness (DCS), though the mechanisms that trigger are still unclear. It has been previously showed that diving may induce increased levels of serum angiotensin converting enzyme. The renin angiotensin aldosterone system (RAAS) is one of the most important regulators of blood pressure and fluid volume. The purpose of the present study was to control the influence of angiotensin II on the appearance of DCS. Methods: Sprague Dawley rats have been pre-treated with inhibitor of angiotensin II receptor type 1 (losartan; 10 mg/kg), angiotensin-converting enzyme (ACE) inhibitor (enalapril; 10 mg/kg), and calcium-entry blocker (nifedipine; 20 mg/kg). The experimental groups were treated for 4 weeks before exposure to hyperbaric pressure while controls were not treated. Seventy-five rats were subjected to a simulated dive at 1000 kPa absolute pressure for 45 min before starting decompression. Clinical assessment took place over a period of 60 min after surfacing. Blood samples were collected for measurements of TBARS, interleukin 6 (IL-6), angiotensin II (ANG II) and ACE. Results: The diving protocol induced 60% DCS in non-treated animals. This ratio was significantly decreased after treatment with enalapril, but not other vasoactive drugs. Enalapril did not change ANG II or ACE concentration, while losartant decreased post dive level of ACE but not ANG II. None of the treatment modified the effect of diving on TBARS and IL-6 values. Conclusion: Results suggests that the rennin angiotensin system is involved in a process of triggering DCS but this has to be further investigated. However, a vasorelaxation mediated process, which potentially could increase the load of inert gas during hyperbaric exposure, and antioxidant properties were excluded by our results.

Evidence of Heritable Determinants of Decompression Sickness in Rats.

INTRODUCTION: Decompression sickness (DCS) is a complex and poorly understood systemic disease caused by inadequate desaturation after a decrease of ambient pressure. Strong variability between individuals is observed for DCS occurrence. This raises questions concerning factors that may be involved in the interindividual variability of DCS occurrence. This study aimed to experimentally assess the existence of heritable factors involved in DCS occurrence by selectively breeding individuals resistant to DCS from a population stock of Wistar rats. METHODS: Fifty-two male and 52 female Wistar rats were submitted to a simulated air dive known to reliably induce about 63% DCS: compression was performed at 100 kPa·min up to 1000 kPa absolute pressure before a 45-min long stay. Decompression was performed at 100 kPa·min with three decompression stops: 5 min at 200 kPa, 5 min at 160 kPa, and 10 min at 130 kPa. Animals were observed for 1 h to detect DCS symptoms. Individuals without DCS were selected and bred to create a new generation, subsequently subjected to the same hyperbaric protocol. This procedure was repeated up to the third generation of rats. RESULTS: As reported previously, this diving profile induced 67% of DCS, and 33% asymptomatic animals in the founding population. DCS/asymptomatic ratio was not initially different between sexes, although males were heavier than females. In three generations, the outcome of the dive significantly changed from 33% to 67% asymptomatic rats, for both sexes. Interestingly, survival in females increased sooner than in males. CONCLUSIONS: This study offers evidence suggesting the inheritance of DCS resistance. Future research will focus on genetic and physiological comparisons between the initial strain and the new resistant population.