The pulmonary oxygen toxicity index.

Pulmonary oxygen toxicity (POT) is a major risk in diving while breathing hyperoxic gas and is also considered in clinical hyperbaric oxygen treatment. The POTindex calculated by the power equation K = t2 × PO24.57 with the recovery form Ktr = Ke × e - [- 0.42 + 0.384 × (PO2)ex] × tr which are based on chemical and physiological principles, have a better prediction power than other suggested approaches. Reduction of vital capacity as well as incidence of POT are well predicted by the POTindex. Both the cumulative pulmonary toxic effect and concomitant recovery were suggested to operate at the lower toxic range of PO2 used in saturation diving K = t2 × PO24.57 × e-0.0135 × t, and further experimental support is supplied. The recovery time constant for the full range of PO2 is presented. POTindex is suggested to replace the old method of UPTD for safe diving. Many diving clubs and diving institutes already adopted the POTindex.

Dipalmitoylphosphatidylcholine in the heart of mice with lupus might support the hypothesis of dual causes of autoimmune diseases.

Lung surfactant dipalmitoylphosphatidylcholine (DPPC) settles on the luminal aspect of blood vessels and forms an active hydrophobic spot - AHS, at which nanobubbles are formed. We hypothesized that large molecules circulating in the blood will adhere and deformed at the gas phase/plasma interface being recognized as autoantigen. NZB mice are afflicted spontaneously with lupus. If their blood vessels contain high levels of DPPC it may support the theory of dual causes of autoimmunity. Phospholipids were extracted from hearts of 8 LPR (lupus) mice and 5 MJP (control mice), and were tested for presence of DPPC. DPPC mg/g was 0.059 in lupus mice and 0.017 in control mice where for equal variance; P = 0.08 and for unequal variance P = 0.048. This trend of 3.5-fold DPPC in lupus mice, supports our hypothesis of dual causes as the origin of autoimmune diseases. The high potential of the hypothesis should be a drive to further explore its validity.

Ex vivo bubble production from ovine large blood vessels: size on detachment and evidence of "active spots".

Nanobubbles formed on the hydrophobic silicon wafer were shown to be the source of gas micronuclei from which bubbles evolved during decompression. Bubbles were also formed after decompression on the luminal surface of ovine blood vessels. Four ovine blood vessels: aorta, pulmonary vein, pulmonary artery, and superior vena cava, were compressed to 1013 kPa for 21 h. They were then decompressed, photographed at 1-s intervals, and bubble size was measured on detachment. There were certain spots at which bubbles appeared, either singly or in a cluster. Mean detachment diameter was between 0.7 and 1.0 mm. The finding of active spots at which bubbles nucleate is a new, hitherto unreported observation. It is possible that these are the hydrophobic spots at which bubbles nucleate, stabilise, and later transform into the gas micronuclei that grow into bubbles. The possible neurological effects of these large arterial bubbles should be further explored.

Evolution of bubbles from gas micronuclei formed on the luminal aspect of ovine large blood vessels.

It has been shown that tiny gas nanobubbles form spontaneously on a smooth hydrophobic surface submerged in water. These nanobubbles were shown to be the source of gas micronuclei from which bubbles evolved during decompression of silicon wafers. We suggest that the hydrophobic inner surface of blood vessels may be a site of nanobubble production. Sections from the right and left atria, pulmonary artery and vein, aorta, and superior vena cava of sheep (n=6) were gently stretched on microscope slides and exposed to 1013 kPa for 18 h. Hydrophobicity was checked in the six blood vessels by advancing contact angle with a drop of saline of 71±19°, with a maximum of about 110±7° (mean±SD). Tiny bubbles ~30 µm in diameter rose vertically from the blood vessels and grew on the surface of the saline, where they were photographed. All of the blood vessels produced bubbles over a period of 80 min. The number of bubbles produced from a square cm was: in the aorta, 20.5; left atrium, 27.3; pulmonary artery, 17.9; pulmonary vein, 24.3; right atrium, 29.5; superior vena cava, 36.4. More than half of the bubbles were present for less than 2 min, but some remained on the saline-air interface for as long as 18 min. Nucleation was evident in both the venous (superior vena cava, pulmonary artery, right atrium) and arterial (aorta, pulmonary vein, left atrium) blood vessels. This newly suggested mechanism of nucleation may be the main mechanism underlying bubble formation on decompression.

Dynamics of gas micronuclei formed on a flat hydrophobic surface, the predecessors of decompression bubbles.

It is a long-standing hypothesis that the bubbles which evolve as a result of decompression have their origin in stable gas micronuclei. In a previous study (Arieli and Marmur, 2011), we used hydrophilic and monolayer-covered hydrophobic smooth silicon wafers to show that nanobubbles formed on a flat hydrophobic surface may be the gas micronuclei responsible for the bubbles that evolve to cause decompression sickness. On decompression, bubbles appeared only on the hydrophobic wafers. The purpose of the present study was to examine the dynamics of bubble evolution. The numbers of bubbles after decompression were greater with increasing hydrophobicity. Bubbles appeared after decompression from 150 kPa, and their density increased with elevation of the exposure pressure (and supersaturation), up to 400 kPa. The normal force of attraction between the hydrophobic surface and the bubble, as determined from the volume of bubbles leaving the surface of the wafer, was 38×10(-5) N and the tangential force was 20×10(-5) N. We discuss the correlation of these results with previous reports of experimental decompression and bubble formation, and suggest to consider appropriate modification of decompression models.

Decompression sickness bubbles: are gas micronuclei formed on a flat hydrophobic surface?

It is a long-standing hypothesis that the bubbles which evolve as a result of decompression have their origin in stable gas micronuclei lodged in hydrophobic crevices, micelles of surface-active molecules, or tribonucleation. Recent findings supported by atomic force microscopy have indicated that tiny, flat nanobubbles form spontaneously on smooth, hydrophobic surfaces submerged in water. We propose that these nanobubbles may be the gas micronuclei responsible for the bubbles that evolve to cause decompression sickness. To support our hypothesis, we used hydrophilic and monolayer-covered hydrophobic smooth silicon wafers. The experiment was conducted in three main stages. Double distilled water was degassed at the low pressure of 5.60 kPa; hydrophobic and hydrophilic silicon wafers were placed in a bowl of degassed water and left overnight at normobaric pressure. The bowl was then placed in the hyperbaric chamber for 15 h at a pressure of 1013 kPa (=90 m sea water). After decompression, bubbles were observed and photographed. The results showed that bubbles only evolved on the hydrophobic surfaces following decompression. There are numerous hydrophobic surfaces within the living body (e.g., in the large blood vessels), which may thus be the sites where nanobubbles that serve as gas micronuclei for bubble evolution following decompression are formed.

Mass spectrometer for respiratory research.

The mass spectrometer is the ideal gas analyser for many applications in respiratory research. A number of companies which manufactured mass spectrometers suitable for use in respiratory research have long since ceased production. Two modifications in the presently manufactured MAX300-LG (Extrel, Pittsburgh, PA, USA) have made it suitable for studying respiratory physiology. The initial step was to bypass the four-port valve and connect the sampling capillary directly to the roughing pump. The second step was to replace the original capillary, which had an internal diameter of 0.4mm, by a 2-m long capillary having an internal diameter of 0.15 mm. The 90% response time was shortened to 85 ms, which is suitable for respiratory research.

Hyperbaric oxygen pretreatment according to the gas micronuclei denucleation hypothesis reduces neurologic deficit in decompression sickness in rats.

During sudden or too rapid decompression, gas is released within supersaturated tissues in the form of bubbles, the cause of decompression sickness. It is widely accepted that these bubbles originate in the tissue from preexisting gas micronuclei. Pretreatment with hyperbaric oxygen (HBO) has been hypothesized to shrink the gas micronuclei, thus reducing the number of emerging bubbles. The effectiveness of a new HBO pretreatment protocol on neurologic outcome was studied in rats. This protocol was found to carry the least danger of oxygen toxicity. Somatosensory evoked potentials (SSEPs) were chosen to serve as a measure of neurologic damage. SSEPs in rats given HBO pretreatment before a dive were compared with SSEPs from rats not given HBO pretreatment and SSEPs from non-dived rats. The incidence of abnormal SSEPs in the animals subjected to decompression without pretreatment (1,013 kPa for 32 min followed by decompression) was 78%. In the pretreatment group (HBO at 304 kPa for 20 min followed by exposure to 1,013 kPa for 33 min and decompression) this was significantly reduced to 44%. These results call for further study of the pretreatment protocol in higher animals.

Combined effect of denucleation and denitrogenation on the risk of decompression sickness in rats.

We previously hypothesized that the number of bubbles emerging on decompression from a dive, and the resultant risk of decompression sickness (DCS), may be reduced by a process whereby effective gas micronuclei that might otherwise have formed bubbles on decompression are shrunk and eliminated. In a procedure defined by us as denucleation, exposure to hyperbaric oxygen (HBO) would result in oxygen replacing the resident gas in the micronuclei, to be subsequently consumed by the mitochondria when the oxygen pressure is reduced. Support for the validity of our hypothesis may be found in our previous studies on the transparent prawn and the reduction of DCS in the rat. In all of these studies, HBO pretreatment was given before supersaturation with inert gas at high pressure. The purpose of the present study was to compare DCS outcome in rats that underwent nitrogen washout (denitrogenation) alone (9 min O(2) at 507 kPa) after exposure to air at high pressure (33 min at 1,266 kPa), and rats treated by both procedures (denitrogenation + denucleation; 8 min of O(2) breathing followed by 5 min air breathing, both at 507 kPa) after high-pressure air exposure. This was done with the same nitrogen load in both groups before the final decompression (a nitrogen pressure of 467 kPa in fatty and 488 kPa in aqueous tissue). Six of 20 rats in the denitrogenation + denucleation group died, compared with 13 in the denitrogenation group (P < 0.03). Three rats in the denitrogenation + denucleation group suffered mild DCS, recovering completely within 2 h of decompression. The present study indicates an advantage in considering both denitrogenation and denucleation before decompression. This may have practical application before escape from a disabled submarine, when aborting a technical dive, or in the preparation of aviators for high altitude.

The effect of over- or underfilling the soda lime canister on CO2 absorption in two closed-circuit oxygen rebreathers.

O2 diving incidents investigated by our laboratory were related to improper filling of the soda lime canister in closed-circuit oxygen rebreathers. We studied the effect of overfilling or underfilling the canister on CO2 absorption using a continuous flow of 5% CO2. With a full canister in the Oxyger 57, CO2 began to rise at 130-160 min, reaching 1% at 240 min and 1.5% at 270 min. Similar results were obtained after a reduction of 100 g in the quantity of soda lime packed into the canister. After reductions of 200, 300 and 400 g, the rise in CO2 concentration occurred earlier as a function of the amount of the reduction. The level of CO2 in the OxyNG 2 began to rise after 250 min with a full canister, reaching 1% at 340 min and 1.5% at 370 min. After a reduction of 100 g there was a delay in the rise of CO2, which reached 1.5% at 390 min. However, when the reduction was 200, 300 and 400 g, the rise in CO2 concentration tended to occur earlier as a function of the amount of the reduction. For both rebreathers, when the quantity of soda lime was reduced by 200 g or more, there was a considerable difference in timing between the two test measurements for each weight reduction, due to variations in channeling. For an excess of soda lime, moderate pressure was applied manually to achieve a full canister plus 300 g in the OxyNG 2. The initial rise in CO2 concentration started early, at 60 min with a full canister plus 300 g compared to 150 min with a full canister; 1% CO2 was reached at 120 min, compared to 210 min with a full canister. As the use of rebreathers becomes increasingly widespread in diving, close attention should be paid to proper filling of the soda lime canister.

Bubble reduction after decompression in the prawn Palaemon elegans by pretreatment with hyperbaric oxygen.

On the theory that bubbles originate from preexisting micronuclei, we previously demonstrated that pretreatment with hyperbaric O2 (HBO2) reduced the number of bubbles in the prawn decompressed from 203 kPa. In the present study, we examined the effect of two HBO2 pretreatment pressures (405 and 709 kPa) on prawns decompressed from a range of pressures between 203-810 kPa. Prawns from the experimental groups were pretreated with O2 at 405 or 709 kPa for 5 min (series A and series B, respectively). Prawns from the control groups were exposed only to air. Following pretreatment, prawns were exposed to air at the desired pressure until saturated with nitrogen, then subjected to rapid decompression and examined under a light microscope. Series A: HBO2 pretreatment at 405 kPa for 5 min significantly reduced the number of bubbles after decompression from 203, 304 and 405 kPa (p < 0.05). The total volume of accumulated gas was not affected by HBO2. Series B: Pretreatment with HBO2 at 709 kPa significantly reduced the number of bubbles after decompression from 203, 304, 507 and 608 kPa (p < 0.05). Total gas volume after decompression from 507 and 608 kPa was reduced as a result of pretreatment with O2. This study demonstrates that HBO2 pretreatment at 405 kPa is sufficient to reduce the number of bubbles that will emerge on decompression from several levels of compression.

Decompression sickness in the rat following a dive on trimix: recompression therapy with oxygen vs. heliox and oxygen.

Trimix (a mixture of helium, nitrogen, and oxygen) has been used in deep diving to reduce the risk of high-pressure nervous syndrome during compression and the time required for decompression at the end of the dive. There is no specific recompression treatment for decompression sickness (DCS) resulting from trimix diving. Our purpose was to validate a rat model of DCS on decompression from a trimix dive and to compare recompression treatment with oxygen and heliox (helium-oxygen). Rats were exposed to trimix in a hyperbaric chamber and tested for DCS while walking in a rotating wheel. We first established the experimental model, and then studied the effect of hyperbaric treatment on DCS: either hyperbaric oxygen (HBO) (1 h, 280 kPa oxygen) or heliox-HBO (0.5 h, 405 kPa heliox 50%-50% followed by 0.5 h, 280 kPa oxygen). Exposure to trimix was conducted at 1,110 kPa for 30 min, with a decompression rate of 100 kPa/min. Death and most DCS symptoms occurred during the 30-min period of walking. In contrast to humans, no permanent disability was found in the rats. Rats with a body mass of 100-150 g suffered no DCS. The risk of DCS in rats weighing 200-350 g increased linearly with body mass. Twenty-four hours after decompression, death rate was 40% in the control animals and zero in those treated immediately with HBO. When treatment was delayed by 5 min, death rate was 25 and 20% with HBO and heliox, respectively.

CNS oxygen toxicity in closed-circuit diving: signs and symptoms before loss of consciousness.

INTRODUCTION: There is a dearth of information regarding CNS oxygen toxicity accidents in closed-circuit oxygen diving. The aims of the present study were to report the sensations and symptoms that accompany CNS oxygen toxicity accidents, and to evaluate whether loss of consciousness can occur without any warning signs. METHODS: We documented 36 CNS oxygen toxicity accidents in closed-circuit oxygen diving. The full accident inquiry included the first report from the diving unit, an interview of the victim and his buddy by the researchers, and an examination of the diving equipment. RESULTS: The symptoms that appeared before termination of a dive, as reported by the victim or his buddy, were as follows (in descending order of frequency): limb convulsions; hyperventilation; difficulty maintaining a steady depth; headache; and visual disturbances. The symptoms that appeared after detachment from the mouthpiece were, in descending order of frequency: headache; loss of consciousness; confusion; weakness; dizziness; and facial muscle twitching and limb convulsions. A high inspired CO2 [mean 4.2 kPa (29.9 mmHg)] was connected with loss of consciousness. No dive was terminated before at least two symptoms (mean 3.4) had been noted a minimum of 5 min before termination. DISCUSSION: Symptoms that are accepted as being related to CNS oxygen toxicity, as well as others such as headache, difficulty maintaining a steady depth, hyperventilation, weakness, and a choking sensation, were more frequent among the O2 accident victims compared with divers who did not interrupt their dives. CONCLUSION: Awareness of any unusual sensation can prevent a potentially dangerous situation from arising.

Effects of nitrogen and helium on CNS oxygen toxicity in the rat.

The contribution of inert gases to the risk of central nervous system (CNS) oxygen toxicity is a matter of controversy. Therefore, diving regulations apply strict rules regarding permissible oxygen pressures (Po(2)). We studied the effects of nitrogen and helium (0, 15, 25, 40, 50, and 60%) and different levels of Po(2) (507, 557, 608, and 658 kPa) on the latency to the first electrical discharge (FED) in the EEG in rats, with repeated measurements in each animal. Latency as a function of the nitrogen pressure was not homogeneous for each rat. The prolongation of latency observed in some rats at certain nitrogen pressures, mostly in the range 100 to 500 kPa, was superimposed on the general trend for a reduction in latency as nitrogen pressure increased. This pattern was an individual trait. In contrast with nitrogen, no prolongation of latency to CNS oxygen toxicity was observed with helium, where an increase in helium pressure caused a reduction in latency. This bimodal response and the variation in the response between rats, together with a possible effect of ambient temperature on metabolic rate, may explain the conflicting findings reported in the literature. The difference between the two inert gases may be related to the difference in the narcotic effect of nitrogen. Proof through further research of a correlation between individual sensitivity to nitrogen narcosis and protection by N(2) against CNS oxygen toxicity in rat may lead to a personal O(2) limit in mixed-gas diving based on the diver sensitivity to N(2) narcosis.

Optimal oxygen pressure and time for reduced bubble formation in the N2-saturated decompressed prawn.

Bubbles that grow during decompression are believed to originate from preexisting gas micronuclei. We showed that pretreatment of prawns with 203 kPa oxygen before nitrogen loading reduced the number of bubbles that evolved on decompression, presumably owing to the alteration or elimination of gas micronuclei (Arieli Y, Arieli R, and Marx A. J Appl Physiol 92: 2596-2599, 2002). The present study examines the optimal pretreatment for this assumed crushing of gas micronuclei. Transparent prawns were subjected to various exposure times (0, 5, 10, 15, and 20 min) at an oxygen pressure of 203 kPa and to 5 min at different oxygen pressures (PO2 values of 101, 151, 203, 405, 608, and 810 kPa), before nitrogen loading at 203 kPa followed by explosive decompression. After the decompression, bubble density and total gas volume were measured with a light microscope equipped with a video camera. Five minutes at a PO2 of 405 kPa yielded maximal reduction of bubble density and total gas volume by 52 and 71%, respectively. It has been reported that 2-3 h of hyperbaric oxygen at bottom pressure was required to protect saturation divers decompressed on oxygen against decompression sickness. If there is a shorter pretreatment that is applicable to humans, this will be of great advantage in diving and escape from submarines.

Modeling pulmonary and CNS O(2) toxicity and estimation of parameters for humans.

The power expression for cumulative oxygen toxicity and the exponential recovery were successfully applied to various features of oxygen toxicity. From the basic equation, we derived expressions for a protocol in which PO(2) changes with time. The parameters of the power equation were solved by using nonlinear regression for the reduction in vital capacity (DeltaVC) in humans: %DeltaVC = 0.0082 x t(2)(PO(2)/101.3)(4.57), where t is the time in hours and PO(2) is expressed in kPa. The recovery of lung volume is DeltaVC(t) = DeltaVC(e) x e(-(-0.42 + 0.00379PO(2))t), where DeltaVC(t) is the value at time t of the recovery, DeltaVC(e) is the value at the end of the hyperoxic exposure, and PO(2) is the prerecovery oxygen pressure. Data from different experiments on central nervous system (CNS) oxygen toxicity in humans in the hyperbaric chamber (n = 661) were analyzed along with data from actual closed-circuit oxygen diving (n = 2,039) by using a maximum likelihood method. The parameters of the model were solved for the combined data, yielding the power equation for active diving: K = t(2) (PO(2)/101.3)(6.8), where t is in minutes. It is suggested that the risk of CNS oxygen toxicity in diving can be derived from the calculated parameter of the normal distribution: Z = [ln(t) - 9.63 +3.38 x ln(PO(2)/101.3)]/2.02. The recovery time constant for CNS oxygen toxicity was calculated from the value obtained for the rat, taking into account the effect of body mass, and yielded the recovery equation: K(t) = K(e) x e(-0.079t), where K(t) and K(e) are the values of K at time t of the recovery process and at the end of the hyperbaric oxygen exposure, respectively, and t is in minutes.

PCO(2) threshold for CNS oxygen toxicity in rats in the low range of hyperbaric PO(2).

Central nervous system (CNS) oxygen toxicity, as manifested by the first electrical discharge (FED) in the electroencephalogram, can occur as convulsions and loss of consciousness. CO(2) potentiates this risk by vasodilation and pH reduction. We suggest that CO(2) can produce CNS oxygen toxicity at a PO(2) that does not on its own ultimately cause FED. We searched for the CO(2) threshold that will result in the appearance of FED at a PO(2) between 507 and 253 kPa. Rats were exposed to a PO(2) and an inspired PCO(2) in 1-kPa steps to define the threshold for FED. The results confirmed our assumption that each rat has its own PCO(2) threshold, any PCO(2) above which will cause FED but below which no FED will occur. As PO(2) decreased from 507 to 456, 405, and 355 kPa, the percentage of rats that exhibited FED without the addition of CO(2) (F(0)) dropped from 91 to 62, to 8 and 0%, respectively. The percentage of rats (F) having FED as a function of PCO(2) was sigmoid in shape and displaced toward high PCO(2) with the reduction in PO(2). The following formula is suggested to express risk as a function of PCO(2) and PO(2) [abstract: see text] where P(50) is the PCO(2)for the half response and N is power. A small increase in PCO(2) at a PO(2) that does not cause CNS oxygen toxicity may shift an entire population into the risk zone. Closed-circuit divers who are CO(2)retainers or divers who have elevated inspired CO(2)are at increased risk of CNS oxygen toxicity.

Use of a mass spectrometer for direct respiratory gas sampling from the hyperbaric chamber.

BACKGROUND: When conducting respiratory gas measurements during hyperbaric chamber research, it is preferable to carry out gas concentration analysis by mass spectrometry. Gas samples for the mass spectrometer are normally taken from a bypass flow exiting the high pressure chamber to the ambient atmosphere. Under these conditions, mixing in the sampling line smoothes the concentration profile, and much of the advantage of low sampling flow is lost. We propose to use a direct sampling method by mass spectrometer that overcomes these deficiencies. METHODS: In the present study, the original high resistance capillary of a QP 9000 mass spectrometer was inserted through the wall of a hyperbaric chamber. Series A: Air and pure nitrogen flowed alternately (1 s each) via the sampling tip of the mass spectrometer. Series B: End expired CO2 from 15 immersed, professional divers exercising at 405 kPa was measured in a screening test for CO2 retention for nitrox diving. RESULTS: There was no difference in the recorded rise time, fall time and plateau reached in the concentration of oxygen at pressures of 101, 202, 303, 405 and 506 kPa. The new sampling method functioned correctly throughout the full-scale experiment, and the recording of end tidal CO2 was more precise than in the conventional method. CONCLUSIONS. Direct sampling of gases from a hyperbaric chamber by the QP 9000 mass spectrometer has many advantages over sampling of the same gases once they are outside the chamber.

Humidity does not affect central nervous system oxygen toxicity.

Central nervous system (CNS) oxygen toxicity can occur as convulsions and loss of consciousness when hyperbaric oxygen is breathed in diving and hyperbaric medical therapy. Lin and Jamieson (J Appl Physiol 75: 1980-1983, 1993) reported that humidity in the inspired gas enhances CNS oxygen toxicity. Because alveolar gas is fully saturated with water vapor, we could not see a cause and effect and surmised that other factors, such as metabolic rate, might be involved. Rats were exposed to 507- and 608-kPa O(2) in dry (31 or 14%) or humid (99%) atmosphere until the appearance of the first electrical discharge preceding the clinical convulsions. Each rat served as its own control. A thermoneutral temperature (28 +/- 0.4 degrees C) yielded resting CO(2) production of 0.81 +/- 0.06 ml x g(-1) x h(-1). Latency to the first electrical discharge was not affected by humidity. At 507-kPa O(2), latency was 23 +/- 0.4 and 22 +/- 0.7 min in dry and humid conditions, respectively, and, at 608-kPa O(2), latency was 15 +/- 4 and 14 +/- 3 min in dry and humid conditions, respectively. When no effects of CO(2) and metabolic rate are present, humidity does not affect CNS oxygen toxicity. Relevance of the findings to diving and hyperbaric therapy is discussed.