Pulmonary nitric oxide in astronauts before and during long-term spaceflight.

Introduction: During exploratory space flights astronauts risk exposure to toxic planetary dust. Exhaled nitric oxide partial pressure (PENO) is a simple method to monitor lung health by detecting airway inflammation after dust inhalation. The turnover of NO in the lungs is dependent on several factors which will be altered during planetary exploration such as gravity (G) and gas density. To investigate the impacts of these factors on normal PENO, we took measurements before and during a stay at the International Space Station, at both normal and reduced atmospheric pressures. We expected stable PENO levels during the preflight and inflight periods, with lower values inflight. With reduced pressure we expected no net changes of PENO. Material and methods: Ten astronauts were studied during the pre-flight (1 G) and inflight (µG) periods at normal pressure [1.0 ata (atmospheres absolute)], with six of them also monitored at reduced (0.7 ata) pressure and gas density. The average observation period was from 191 days before launch until 105 days after launch. PENO was measured together with estimates of alveolar NO and the airway contribution to the exhaled NO flux. Results: The levels of PENO at 50 mL/s (PENO50) were not stable during the preflight and inflight periods respectively but decreased with time (p = 0.0284) at a rate of 0.55 (0.24) [mean (SD)] mPa per 180 days throughout the observation period, so that there was a significant difference (p < 0.01, N = 10) between gravity conditions. Thus, PENO50 averaged 2.28 (0.70) mPa at 1 G and 1.65 (0.51) mPa during µG (-27%). Reduced atmospheric pressure had no net impact on PENO50 but increased the airway contribution to exhaled NO. Discussion: The time courses of PENO50 suggest an initial airway inflammation, which gradually subsided. Our previous hypothesis of an increased uptake of NO to the blood by means of an expanded gas-blood interface in µG leading to decreased PENO50 is neither supported nor contradicted by the present findings. Baseline PENO50 values for lung health monitoring in astronauts should be obtained not only on ground but also during the relevant gravity conditions and before the possibility of inhaling toxic planetary dust.

Lung diffusing capacity for nitric oxide in space: microgravity gas density interactions.

Introduction: During manned space exploration lung health is threatened by toxic planetary dust and radiation. Thus, tests such as lung diffusing capacity (DL) are likely be used in planetary habitats to monitor lung health. During a DL maneuver the rate of uptake of an inspired blood-soluble gas such as nitric oxide (NO) is determined (DLNO). The aim of this study was to investigate the influence of altered gravity and reduced atmospheric pressure on the test results, since the atmospheric pressure in a habitat on the moon or on Mars is planned to be lower than on Earth. Changes of gravity are known to alter the blood filling of the lungs which in turn may modify the rate of gas uptake into the blood, and changes of atmospheric pressure may alter the speed of gas transport in the gas phase. Methods: DLNO was determined in 11 subjects on the ground and in microgravity on the International Space Station. Experiments were performed at both normal (1.0 atm absolute, ata) and reduced (0.7 ata) atmospheric pressures. Results: On the ground, DLNO did not differ between pressures, but in microgravity DLNO was increased by 9.8% (9.5) (mean [SD]) and 18.3% (15.8) at 1.0 and 0.7 ata respectively, compared to normal gravity, 1.0 ata. There was a significant interaction between pressure and gravity (p = 0.0135). Discussion: Estimates of the membrane (DmNO) and gas phase (DgNO) components of DLNO suggested that at normal gravity a reduced pressure led to opposing effects in convective and diffusive transport in the gas phase, with no net effect of pressure. In contrast, a DLNO increase with reduced pressure at microgravity is compatible with a substantial increase of DmNO partially offset by reduced DgNO, the latter being compatible with interstitial edema. In microgravity therefore, DmNO would be proportionally underestimated from DLNO. We also conclude that normal values for DL in anticipation of planetary exploration should be determined not only on the ground but also at the gravity and pressure conditions of a future planetary habitat.

Short-arm centrifugation as a partially effective musculoskeletal countermeasure during 5-day head-down tilt bed rest--results from the BRAG1 study.

PURPOSE: Human centrifugation, also called artificial gravity (AG), is proposed as a combined strategy against detrimental effects of microgravity in long-term space missions. This study scrutinized human short-arm centrifugation as countermeasure against musculoskeletal de-conditioning. METHOD: Eleven healthy male subjects [mean age of 34 (SD 7) years] completed the cross-over trial, including three campaigns of -6° head-down tilt bed rest (HDT) for 5 days, with preceding baseline data collection and recovery phases. Bed rest without AG was used as control condition (Ctrl), and AG with 1 g at the center of mass applied once per day for 30 min in one bout (AG1×30) and in 6 bouts of 5 min (AG6×5, 3-min rest between bouts) as experimental conditions. End-points were muscle strength, vertical jump performance, and biomarkers of bone and protein metabolism. RESULT: AG6×5 was better tolerated than AG1×30. Bone resorption markers CTX, NTX, and DPD all increased by approximately 25 % toward the end of bed rest (P < 0.001), and nitrogen balance decreased by approximately 3 g/day (P < 0.001), without any protection by AG (P > 0.4). Decreases in vertical jump height by 2.1 (SE 0.6) cm after Ctrl bed rest was prevented by either of the AG protocols (P = 0.039). CONCLUSION: The present study yielded succinct catabolic effects upon muscle and bone metabolism that were un-prevented by AG. The preservation of vertical jump performance by AG in this study is likely caused by central nervous rather than by peripheral musculoskeletal effects.

Effects of an artificial gravity countermeasure on orthostatic tolerance, blood volumes and aerobic power after short-term bed rest (BR-AG1).

Exposure to artificial gravity (AG) in a short-arm centrifuge has potential benefits for maintaining human performance during long-term space missions. Eleven subjects were investigated during three campaigns of 5 days head-down bed rest: 1) bed rest without countermeasures (control), 2) bed rest and 30 min of AG (AG1) daily, and 3) bed rest and six periods of 5 min AG (AG2) daily. During centrifugation, the supine subjects were exposed to AG in the head-to-feet direction with 1 G at the center of mass. Subjects participated in the three campaigns in random order. The cardiovascular effects of bed rest and countermeasures were determined from changes in tolerance to a head-up tilt test with superimposed lower body negative pressure (HUT), from changes in plasma volume (PV) and from changes in maximum aerobic power (V̇o2 peak) during upright work on a cycle ergometer. Complete data sets were obtained in eight subjects. After bed rest, HUT tolerance times were 36, 64, and 78% of pre-bed rest baseline during control, AG1 and AG2, respectively, with a significant difference between AG2 and control. PV and V̇o2 peak decreased to 85 and 95% of pre-bed rest baseline, respectively, with no differences between the treatments. It was concluded that the AG2 countermeasure should be further investigated during future long-term bed rest studies, especially as it was better tolerated than AG1. The superior effect of AG2 on orthostatic tolerance could not be related to concomitant changes in PV or aerobic power.

Lung diffusing capacity for nitric oxide at lowered and raised ambient pressures.

Lung diffusing capacity for NO (DLNO) was determined in eight subjects at ambient pressures of 505, 1015, and 4053hPa (379, 761 and 3040mmHg) as they breathed normoxic gases. Mean values were 116.9±11.1 (SEM), 113.4±11.1 and 99.3±10.1mlmin(-1)hPa(-1)at 505, 1015, and 4053hPa, with a 13% difference between the two higher pressures (P=0.017). The data were applied to a model with two serially coupled conductances; the gas phase (DgNO, variable with pressure), and the alveolo-capillary membrane (DmNO, constant). The data fitted the model well and we conclude that diffusive transport of NO in the peripheral lung is inversely related to gas density. At normal pressure DmNO was approximately 5% larger than DLNO, suggesting that the Dg factor then is not negligible. We also conclude that the density of the breathing gas is likely to impact the backdiffusion of naturally formed NO from conducting airways to the alveoli.

Effects of ambient pressure on pulmonary nitric oxide.

Airway nitric oxide (NO) has been proposed to play a role in the development of high-altitude pulmonary edema. We undertook a study of the effects of acute changes of ambient pressure on exhaled and alveolar NO in the range 0.5-4 atmospheres absolute (ATA, 379-3,040 mmHg) in eight healthy subjects breathing normoxic nitrogen-oxygen mixtures. On the basis of previous work with inhalation of low-density helium-oxygen gas, we expected facilitated backdiffusion and lowered exhaled NO at 0.5 ATA and the opposite at 4 ATA. Instead, the exhaled NO partial pressure (Pe(NO)) did not differ between pressures and averaged 1.21 ± 0.16 (SE) mPa across pressures. As a consequence, exhaled NO fractions varied inversely with pressure. Alveolar estimates of the NO partial pressure differed between pressures and averaged 88 (P = 0.04) and 176 (P = 0.009) percent of control (1 ATA) at 0.5 and 4 ATA, respectively. The airway contribution to exhaled NO was reduced to 79% of control (P = 0.009) at 4 ATA. Our finding of the same Pe(NO) at 0.5 and 1 ATA is at variance with previous findings of a reduced Pe(NO) with inhalation of low-density gas at normal pressure, and this discrepancy may be due to the much longer durations of low-density gas breathing in the present study compared with previous studies with helium-oxygen breathing. The present data are compatible with the notion of an enhanced convective backtransport of NO, compensating for attenuated backdiffusion of NO with increasing pressure. An alternative interpretation is a pressure-induced suppression of NO formation in the airways.

Formation of new bioactive organic nitrites and their identification with gas chromatography-mass spectrometry and liquid chromatography coupled to nitrite reduction.

Nitric oxide (NO) donors, notably organic nitrates and nitrites are used therapeutically but tolerance develops rapidly, making the use of e.g. nitroglycerin difficult. NO donation in the pulmonary vascular bed might be useful in critically ill patients. Organic nitrites are not associated with tachyphylaxis but may induce methaemoglobinemia and systemic hypotension which might hamper their use. We hypothesised that new lung-selective NO donors can be identified by utilizing exhaled NO as measure for pulmonary NO donation and systemic arterial pressure to monitor hypotension and tolerance development. Solutions of alcohols and carbohydrates were reacted with NO gas and administered to ventilated rabbits for evaluation of in vivo NO donation. Chemical characterization was made by liquid chromatography with on-line nitrite reduction (LC-NO) and by gas chromatography-mass spectrometry (GC-MS). In vivo experiments showed that the hydroxyl-containing compounds treated with NO gas yielded potent NO donors, via nitrosylation to organic nitrites. Analyses by LC-NO showed that the reaction products were able to release NO in vitro. In GC-MS the reaction products were determined to be the organic nitrites, where some are new chemical entities. Non-polar donors preferentially increased exhaled NO with less effect on systemic blood pressure whereas more polar molecules had larger effects on systemic blood pressure and less on exhaled NO. We conclude that new organic nitrites suitable for intravenous administration are produced by reacting NO gas and certain hydroxyl-containing compounds in aqueous solutions. Selectivity of different organic nitrites towards the pulmonary and systemic circulation, respectively, may be determined by molecular polarity.

Effects of acceleration in the Gz axis on human cardiopulmonary responses to exercise.

The aim of this paper was to develop a model from experimental data allowing a prediction of the cardiopulmonary responses to steady-state submaximal exercise in varying gravitational environments, with acceleration in the G(z) axis (a (g)) ranging from 0 to 3 g. To this aim, we combined data from three different experiments, carried out at Buffalo, at Stockholm and inside the Mir Station. Oxygen consumption, as expected, increased linearly with a (g). In contrast, heart rate increased non-linearly with a (g), whereas stroke volume decreased non-linearly: both were described by quadratic functions. Thus, the relationship between cardiac output and a (g) was described by a fourth power regression equation. Mean arterial pressure increased with a (g) non linearly, a relation that we interpolated again with a quadratic function. Thus, total peripheral resistance varied linearly with a (g). These data led to predict that maximal oxygen consumption would decrease drastically as a (g) is increased. Maximal oxygen consumption would become equal to resting oxygen consumption when a (g) is around 4.5 g, thus indicating the practical impossibility for humans to stay and work on the biggest Planets of the Solar System.

Effect of blood redistribution on exhaled and alveolar nitric oxide: a hypergravity model study.

Alveolar (CA(NO)) and exhaled nitric oxide (FE(NO)) concentrations, mainly regarded as inflammation surrogates, may also be affected by perfusion redistribution changing alveolar transfer factor (DA(NO)). A model of blood redistribution is hypergravity, Karlsson et al. (2009b) found, at 2G, increases of 22% and 70%, for FE(NO), and CA(NO), respectively. The present study aimed at theoretically estimating the amplitude of DA(NO) changes that mimic these experimental data. An equation describing convection, diffusion and NO sources was solved in a 2-trumpet model (parallel dependent and non-dependent lung units). Acinar airways lumen reduction was also simulated. A reduction of 33% of the overall DA(NO) (-51% in the non-dependent unit) along with a 36% reduction of acinar airways lumen reproduced experimental findings. In conclusion, substantial FE(NO) and CA(NO) increases may be accounted for by a decrease of the alveolo-capillaries contact surface, here hypergravity-induced. Acinar airway constriction may also have a part in the overall FE(NO) increase.

Determinants of oxygen consumption during exercise on cycle ergometer: the effects of gravity acceleration.

The hypothesis that changes in gravity acceleration (a(g)) affect the linear relationships between oxygen consumption VO2 and mechanical power (w ) so that at any w, VO2 increases linearly with a(g) was tested under conditions where the weight of constant-mass legs was let to vary by inducing changes in a(g) in a human centrifuge. The effects of a(g) on the VO2/w relationship were studied on 14 subjects at two pedalling frequencies (f(p), 1.0 and 1.5 Hz), during four work loads on a cycle ergometer (25, 50, 75 and 100 W) and at four a(g) levels (1.0, 1.5, 2.0 and 2.5 times normal gravity). VO2 increased linearly with w. The slope did not differ significantly at various a(g) and f(p), suggesting invariant mechanical efficiency during cycling, independent of f(p) and a(g). Conversely, the y-intercept of the VO2/w relationship, defined as constant b, increased linearly with a(g). Constant b is the sum of resting VO2 plus internal metabolic power (E (i)). Since the former was the same at all investigated a(g), the increase in constant b was entirely due to an increase in E (i). Since the VO2 versus w lines had similar slopes, the changes in E (i) entirely explained the higher VO2 at each w, as a(g) was increased. In conclusion, the effects of a(g) on VO2 are mediated through changes in E (i), and not in w or in resting VO2.

Microgravity decreases and hypergravity increases exhaled nitric oxide.

Inhalation of toxic dust during planetary space missions may cause airway inflammation, which can be monitored with exhaled nitric oxide (NO). Gravity will differ from earth, and we hypothesized that gravity changes would influence exhaled NO by altering lung diffusing capacity and alveolar uptake of NO. Five subjects were studied during microgravity aboard the International Space Station, and 10 subjects were studied during hypergravity in a human centrifuge. Exhaled NO concentrations were measured during flows of 50 (all gravity conditions), 100, 200, and 500 ml/s (hypergravity). During microgravity, exhaled NO fell from a ground control value of 12.3 +/- 4.7 parts/billion (mean +/- SD) to 6.6 +/- 4.4 parts/billion (P = 0.016). In the centrifuge experiments and at the same flow, exhaled NO values were 16.0 +/- 4.3, 19.5 +/- 5.1, and 18.6 +/- 4.7 parts/billion at one, two, and three times normal gravity, where exhaled NO in hypergravity was significantly elevated compared with normal gravity (P

Lower exhaled nitric oxide in hypobaric than in normobaric acute hypoxia.

Analysis of exhaled nitric oxide (NO) has become an accepted complementary tool in the management of inflammatory airway diseases. Previous studies have demonstrated reduced exhaled NO at altitude and ascribed their findings to hypoxia. We studied exhaled NO partial pressures (Pe(NO)) in eight healthy subjects at reduced ambient pressure down to 540 hPa (equivalent to 5000 m altitude) and at sea level, with equivalently hypoxic breathing gases (down to 11.3% O2 in N2). Pe(NO) readings were corrected for gas density effects on the instrument performance. Sea level control values for Pe(NO) at an exhaled flow of 50 mls(-1) averaged 2.4 mPa and were virtually unchanged with normobaric hypoxia down to an inspired P(O)(2) of 10.7 kPa. With the same degree of hypoxia, hypobaric Pe(NO) was 1.4 mPa. The reduction in hypobaric Pe(NO) of up to 33+/-16% (mean+/-SD) in comparison to normobaric Pe(NO), is likely to have been caused by enhanced axial back diffusion of NO because of the reduced gas density and an associated increased alveolar NO uptake to the blood.

Measuring exhaled nitric oxide at high altitude.

The altitude performance of two NO analysers using different NO detectors was studied. The analysers and their flow regulators were tested with simulated exhalations of reference gases. At 4000 m, volume flow was +35% and mass flow -24% of nominal in both instruments. The reduced mass flow increased the exhaled NO fraction by 26% for a given rate of NO excretion. Furthermore, the electrochemical NO detector in one analyser showed an increased signal level for a given partial pressure of test gas. Taken together, these two effects increased the signal output by 60% in comparison to the NO partial pressure. To avoid the above errors, it is proposed that the flow regulator should be readjusted to give a volume flow of 50 ml s(-1) at the altitude of interest and that the analyser should be recalibrated to the operational altitude. Finally, it is recommended that exhaled NO should always be reported as partial pressure and not as volume fraction, in order to compare measurements at any altitude.

Venous gas emboli and exhaled nitric oxide with simulated and actual extravehicular activity.

The decompression experienced due to the change in pressure from a space vehicle (1013hPa) to that in a suit for extravehicular activity (EVA) (386hPa) was simulated using a hypobaric chamber. Previous ground-based research has indicated around a 50% occurrence of both venous gas emboli (VGE) and symptoms of decompression illness (DCI) after similar decompressions. In contrast, no DCI symptoms have been reported from past or current space activities. Twenty subjects were studied using Doppler ultrasound to detect any VGE during decompression to 386hPa, where they remained for up to 6h. Subjects were supine to simulate weightlessness. A large number of VGE were found in one subject at rest, who had a recent arm fracture; a small number of VGE were found in another subject during provocation with calf contractions. No changes in exhaled nitric oxide were found that can be related to either simulated EVA or actual EVA (studied in a parallel study on four cosmonauts). We conclude that weightlessness appears to be protective against DCI and that exhaled NO is not likely to be useful to monitor VGE.

Regional lung blood flow and ventilation in upright humans studied with quantitative SPECT.

We used quantitative Single Photon Emission Computed Tomography (SPECT) to study the effect of the upright posture on regional lung blood flow and ventilation. Nine (upright) plus seven (prone and supine) healthy volunteers were studied awake, breathing spontaneously. Regional blood flow and ventilation were marked in sitting upright, supine and prone postures using (113m)In-labeled macroaggregates and inhaled Technegas ((99m)Tc); both remain fixed in the lung after administration. All images were obtained while supine. In comparison with horizontal postures, both blood flow and ventilation were greater in caudal regions when upright. The redistribution was greater for blood flow than for ventilation, resulting in decreasing ventilation-to-perfusion ratios down the lung when upright. We conclude that gravity redistributes regional blood flow and ventilation in the upright posture, while the influence is much less in the supine and prone postures.

Central command and metaboreflex cardiovascular responses to sustained handgrip during microgravity.

Four subjects were studied before and during a 16-day space flight. The test included 2min of rest, 2min of sustained handgrip (SHG), and 2min of post-exercise circulatory occlusion (PECO). Heart rate (HR) and mean arterial pressure (MAP) responses to central command and mechanoreceptor stimulation were determined from the difference between SHG and PECO. Responses to metaboreceptor stimulation were determined from the difference between PECO and rest. Late in-flight (days 12-14) the central command/mechanoreceptor component of the HR response was reduced by 5bpm (P=0.01) from its pre-flight value of 15 (+/-3)bpm (mean (+/-SEM)). At the same time the metaboreflex responses of HR and MAP were unchanged. The attenuated HR response to central command was likely of baroreflex origin. Together with a parallel study of PECO after dynamic leg exercise, our data indicate that central processing of metaboreflex inputs is unchanged in microgravity whereas metaboreflex inputs from weight-bearing muscles are enhanced.

Influence of combined exercise and gravity transients and apnea on hemodynamics.

Hemodynamic responses to combined heavy dynamic leg exercise (hiP), breath holding (BH) and gravity-induced blood volume shifts direction were studied. Thirteen subjects were studied at normal gravity and 12 during parabolic flight, performing 20 s hiP or combined hiP&BH (stimulus period) from a baseline of 30 W at normal gravity (1 G(z+)). Heart rate and mean arterial pressure responses to BH were similar between gravity conditions, but stroke volume (SV) differed markedly between gravity conditions: at 1 G(z+) SV was higher [112 +/- 16 ml (mean +/- SD)] during BH, than during eupnea [101 +/- 17 ml (P < 0.05, N = 13)]. In weightlessness the corresponding SV values were 105 +/- 16 and 127 +/- 20 ml, respectively (P < 0.05, N = 6). Transthoracic electrical conductance (TTC) was used as index for intrathoracic volume. TTC fell significantly during BH. This decrease was attenuated in weightlessness. It is concluded that the transient microgravity temporarily reduces the efficiency of the muscle pump so that the deep inspiration at the onset of the high-intensity exercise and breath-hold period cannot augment venous return as it could during identical manoeuvres at normal gravity.