Gain-of-function Tibetan PHD2D4E;C127S variant suppresses monocyte function: A lesson in inflammatory response to inspired hypoxia.

BACKGROUND: We have previously described an evolutionarily selected Tibetan prolyl hydroxylase-2 (PHD2D4E;C127S) variant that degrades the hypoxia-inducible factor (HIFα) more efficiently and protects these highlanders from hypoxia-triggered elevation in haemoglobin concentration. High altitude is known to cause acute mountain sickness (AMS) and high-altitude pulmonary edema (HAPE) in a section of rapidly ascending non-acclimatised lowlanders. These morbidities are often accompanied by inflammatory response and exposure to hypobaric hypoxia is presumed to be the principal causative agent. We have investigated whether PHD2D4E;C127S variant is associated with prevention of hypoxia-mediated inflammatory milieu in Tibetan highlanders and therefore identify a potential target to regulate inflammation. METHODS: We genotyped the Tibetans using DNA isolated from whole blood. Thereafter immunophenotying was performed on PBMCs from homozygous PHD2D4E;C127S and PHD2WT individuals using flow cytometry. RNA isolated from these individuals was used to evaluate the peripheral level of important transcripts associated with immune as well as hypoxia response employing the nCounter technology. The ex-vivo findings were validated by generating monocytic cell lines (U937 cell line) expressing PHD2D4E;C127S and PHD2WT variants post depletion of endogenous PHD2. We had also collected whole blood samples from healthy travellers and travellers afflicted with AMS and HAPE to evaluate the significance of our ex-vivo and in vitro findings. Hereafter, we also attempted to resolve hypoxia-triggered inflammation in vitro as well as in vivo by augmenting the function of PHD2 using alpha-ketoglutarate (αKG), a co-factor of PHD2. FINDINGS: We report that homozygous PHD2D4E;C127S highlanders harbour less inflammatory and patrolling monocytes in circulation as compared to Tibetan PHD2WT highlanders. In response to in vitro hypoxia, secretion of IL6 and IL1ß from PHD2D4E;C127S monocytes, and their chemotactic response compared to the PHD2WT are compromised, corresponding to the down-modulated expression of related signalling molecules RELA, JUN, STAT1, ATF2 and CXCR4. We verified these functional outcomes in monocytic U937 cell line engineered to express PHD2D4E;C127S and confirmed the down-modulation of the signalling molecules at protein level under hypoxia. In contrast, non-Tibetan sojourners with AMS and HAPE at high altitude (3,600 m above sea level) displayed significant increase in these inflammatory parameters. Our data henceforth underline the role of gain-of-function of PHD2 as the rate limiting factor to harness hyper-activation of monocytes in hypoxic environment. Therefore upon pre-treatment with αKG, we observed diminished inflammatory response of monocytes in vitro and reduction in leukocyte infiltration to the lungs in mice exposed to normobaric hypoxia. INTERPRETATION: Our report suggests that gain-of-function PHD2 D4E;C127S variant can therefore protect against inflammation elicited by hypobaric hypoxia. Augmentation of PHD2 activity therefore may be an important method to alleviate inflammatory response to inspired hypoxia. FUNDING: This study is supported by the Department of Biotechnology, Government of India.

The effect of zolpidem on cognitive function and postural control at high altitude.

Study Objectives: Sleep is altered at high altitude leading many mountaineers to use hypnotics in order to improve sleep efficiency. While after a full night at altitude the short-acting hypnotic zolpidem does not appear to alter cognitive function, residual adverse effects should be considered following early waking-up as performed by mountaineers. We hypothesized that zolpidem intake at high altitude would alter cognitive function 4 hours after drug intake. Methods: In a randomized double-blind controlled cross-over study, 22 participants were evaluated during two nights at sea level and two nights at 3800 m, 4 hours after zolpidem (10 mg) or placebo intake at 10:00 pm. Polygraphic recording was performed until waking-up at 01:30 am. Sleep quality, sleepiness and symptoms of acute mountain sickness were assessed by questionnaires. Two cognitive tasks (Simon task and duration-production task) were performed at rest and during exercise and postural control was evaluated. Results: Zolpidem increased reaction time in all conditions (zolpidem 407 ± 9 ms vs. placebo 380 ± 11 ms; p < 0.001) and error rate in incongruent trials only (10.2 ± 1.1% vs. 7.8 ± 0.8%; p < 0.01) in the Simon task and increased time perception variability (p < 0.001). Zolpidem also altered postural parameters (e.g. center of pressure area, zolpidem 236 ± 171.5 mm2 vs. placebo 119.6 ± 59 mm2; p < 0.001). Zolpidem did not affect apnea-hypopnea index and mean arterial oxygen saturation (p > 0.05) but increased sleep quality (p < 0.001). Zolpidem increased symptoms of acute mountain sickness and sleepiness (p < 0.05). Conclusions: Acute zolpidem intake at high altitude alters cognitive functions and postural control during early wakening which may be deleterious for safety and performances of climbers.

Hormonal contraceptives and travel to high altitude.

Women frequently ask about the safety and efficacy of using hormonal contraception (HC), either oral contraceptive pills (OC) or other forms, when traveling to high altitude locales. What are the risks and benefits of using HC at high altitude? Does HC affect acclimatization, exercise performance, or occurrence of acute mountain sickness? This article reviews current data regarding the risks and benefits of HC at high altitude, both demonstrated and theoretical, with the aim of helping health care providers to advise women traveling above 2500 meters. Most healthy women can safely use HC when traveling to high altitude, but should be aware of the potential risks and inconveniences.

Oral contraceptive use and acute mountain sickness in South Pole workers.

INTRODUCTION: Progesterone has a number of properties that could influence the development of acute mountain sickness (AMS), including anti-inflammation, respiratory smooth muscle relaxation, ventilatory stimulation, and antidiuretic characteristics. Oral contraceptive (OC) use decreases levels of circulating progesterone by preventing ovulation. We hypothesized rates of AMS development would be significantly higher in OC users as compared to Non-OC users in a population traveling rapidly to the South Pole. METHODS: There were 50 female subjects (OC N = 13, no OC N = 37) who traveled by airplane from Sea Level (SL) to Altitude (ALTD) (-3200 m) in < 4 h and were monitored for the development of AMS. SL and ALTD measurements of anthropometrics, vital signs, hematologic variables, blood chemistries, electrolytes, endocrine responses, and pulmonary function were assessed with t-test and Chi-square analyses, P < 0.05. RESULTS: As compared to Non-OC users, OC users had lower progesterone levels (ng x ml(-1)) at SL (0.7 +/- 0.5 vs. 3.2 +/- 4.6) and at ALTD (0.7 +/- 0.7 vs. 3.1 +/- 4.6). AMS was significantly more prevalent in OC users (85%) as compared to Non-OC users (51%). Acetazolamide prophylaxis was not protective, with a greater proportion of OC users (100%) developing AMS despite its use as compared to Non-OC users (50%). Blood pressure responses also differed significantly, with OC users displaying higher mean arterial pressures at ALTD vs. Non-OC users. CONCLUSION: OC use at ALTD is associated with an increased risk for the development of AMS. Acetazolamide prophylaxis with OC use was also associated with an increased rate of AMS development.

Corticosterone synthesis inhibitor metyrapone ameliorates chronic hypobaric hypoxia induced memory impairment in rat.

Chronic exposure to hypobaric hypoxia causes oxidative stress and neurodegeneration leading to memory impairment. The present study aimed at investigating the role of corticosterone in hypoxia induced neurodegeneration and effect of metyrapone, a corticosterone synthesis inhibitor that reduces the stress induced elevation of corticosterone without affecting the basal level, in ameliorating chronic hypobaric hypoxia induced cognitive decline. Rats were exposed to simulated altitude of 25,000 ft for 0, 3, 7, 14 and 21 days to determine the temporal alterations in corticosterone and its receptors following exposure to hypobaric hypoxia. Our results showed an elevation of corticosterone in plasma and hippocampal tissue following 7 days of exposure, which declined on prolonged hypoxic exposure for 21 days. A concomitant increase in ROS and lipid peroxidation was observed along with depletion of intracellular antioxidants. Glucocorticoid and mineralocorticoid receptors were upregulated on 3 and 7 days of hypoxic exposure. Though expression of Glut1 and Glut3 were upregulated on 3 days of hypoxic exposure, sharp decline in Glut1 expression following 7 days of hypoxic exposure leads to reduced neuronal glucose uptake. Administration of metyrapone from 3rd to 7th day of hypoxic exposure to suppress hypoxia induced increase in corticosterone levels resulted in reduced oxidative damage, neurodegeneration and improvement of intracellular energy status. The metyrapone treated hypoxic animals performed better in the Morris Water Maze. Further, administration of exogenous corticosterone along with metyrapone during hypoxic exposure blunted the neuroprotective effect of metyrapone indicating a role for corticosterone in mediating hypobaric hypoxia induced neurodegeneration and memory impairment.

Severe acute mountain sickness and suspect high altitude cerebral edema related to nitroglycerin use.

An elite mountaineer reported severe acute mountain sickness and ataxia during an 8000-m expedition and concomitant use of transdermal nitroglycerin patches aimed to prevent frostbites. Use of nitroglycerin for this purpose is off-label, and its safety has not been assessed. Moreover, a relation between nitrate-induced cerebral vasodilation and high altitude cerebral edema is theoretically possible on a pathophysiological basis. It is our opinion that nitroglycerin use at high altitude should be discouraged, as efficacy in the prevention of frostbites is questionable and safety has not been assessed.

Oral contraceptives, exercise, and acute mountain sickness in women.

BACKGROUND: Previous research has found that exercise exacerbated acute mountain sickness (AMS) in men. PURPOSE: The current study tested this relationship in women taking oral contraceptives. METHODS: We studied seven women at 428 mmHg for 10 h; once while at rest (R) and once while performing intermittent exercise (EX). RESULTS: AMS scores had a slight increase at 9 vs. 0 h at altitude in both trials (p < 0.05). Resting measurements of ventilation (VE), arterial oxygen saturation (SPO2), end tidal O2 (PETO2), and end tidal CO2 (PETCO2) were not different over time or between trials (p > 0.05). While fluid intake did not change, urine output increased during the 0-3 h period, regardless of trial, and returned to baseline values by the 6-9 h period (218 +/- 37 vs. 121 +/- 22 ml x h(-1); p < 0.05). During exercise, SPO2 significantly dropped compared with similar time points in R (73.1 +/- 1.1 vs. 85.7 +/- 1.8%; p < 0.05). Despite exercise-induced desaturation, the AMS scores were not significantly different between R and EX. CONCLUSION: These results suggest that oral contraceptives may cause a compensation for the physiological responses to exercise critical for the development of AMS.