Head-down Tilt Bed Rest Study Research Articles

State of the Retina and Optic Nerve in 21-Day Head-Down Tilt Bed Rest

Studies in conditions of head-down tilt bed rest are widely used experiments which imitate redistribution of body fluids similar to the redistribution of fluid media in microgravity. To study the mechanism of development of spaceflight-associated neuro-ocular syndrome (SANS) in head-down tilt bed rest studies scientists have evaluated, among other things, participants’ visual system parameters. The main symptom of SANS is an edema of an optic disc and a retina. To date, the results describing the influence of head-down tilt bed rest on the visual system are ambiguous, which determines the relevance of further research. The aim of the study is to analyze retinal morphometry in the optic disc area and macular area in participants before and after 21-day head-down tilt bed rest. The study was performed using an optical coherence tomography with angiography function (Optovue RTVue XR Avanti System) before and after 21-days of head-down tilt bed rest. In 4 participants (8 eyes) (men, mean age ± standard deviation: 29.3 ± 3.9 years) the macular zone and optic disc area were evaluated according to standard scanning modes. To analyze the values obtained, we used mean tendency scores, mean comparison criteria, and graphical analysis. The results of the retinal thickness in the optic disc zone were consistent with those of other researchers, indicating a possible increase in retinal thickness, but our results did not reach the level of statistical significance. A statistically significant thickening of the retina in the macular zone was shown for the first time, but the increase is significantly less in amplitude than the thickening of the nerve fiber layer in the optic disc zone. Analysis of vascular density under head-down tilt bed rest conditions was performed for the first time both for the optic disc zone and the macula zone, but additional studies are required to obtain steady conclusions. For the first time, an increase in retinal thickness in the macula zone in participants of head-down tilt bed rest experiments was shown; for the optic disc zone, data consistent with the results of other researchers were obtained. For the first time, angiographic data were obtained for the macula and optic disk areas, but the obtained differences in vascular density did not reach the level of statistical significance.

Mechanical deconditioning of the heart due to long-term bed rest as observed on seismocardiogram morphology

During head-down tilt bed rest (HDT) the cardiovascular system is subject to headward fluid shifts. The fluid shift phenomenon is analogous to weightlessness experienced during spaceflight microgravity. The purpose of this study was to investigate the effect of prolonged 60-day bed rest on the mechanical performance of the heart using the morphology of seismocardiography (SCG). Three-lead electrocardiogram (ECG), SCG and blood pressure recordings were collected simultaneously from 20 males in a 60-day HDT study (MEDES, Toulouse, France). The study was divided into two campaigns of ten participants. The first commenced in January, and the second in September. Signals were recorded in the supine position during the baseline data collection (BDC) before bed rest, during 6° HDT bed rest and during recovery (R), post-bed rest. Using SCG and blood pressure at the finger, the following were determined: Pulse Transit Time (PTT); and left-ventricular ejection time (LVET). SCG morphology was analyzed using functional data analysis (FDA). The coefficients of the model were estimated over 20 cycles of SCG recordings of BDC12 and HDT52. SCG fiducial morphology AO (aortic valve opening) and AC (aortic valve closing) amplitudes showed significant decrease between BDC12 and HDT52 (p < 0.03). PTT and LVET were also found to decrease through HDT bed rest (p < 0.01). Furthermore, PTT and LVET magnitude of response to bed rest was found to be different between campaigns (p < 0.001) possibly due to seasonal effects on of the cardiovascular system. Correlations between FDA and cardiac timing intervals PTT and LVET using SCG suggests decreases in mechanical strength of the heart and increased arterial stiffness due to fluid shifts associated with the prolonged bed rest.

Visual Attention Relates to Operator Performance in Spacecraft Docking Training.

BACKGROUND: Manually controlled docking of a spacecraft to a space station is an operational task that poses high demands on cognitive and perceptual functioning. Effective processing of visual information is crucial for success. Eye tracking can reveal the operator's attentional focus unobtrusively and objectively. Therefore, our aim was to test the feasibility of eye tracking during a simulation of manual docking and to identify links between visual information processing and performance.METHODS: We hypothesized that duration and number of gazes to specific regions of interest of the simulation (total dwell time and number of dwells) would be associated with docking accuracy. Eye movements were recorded in 10 subjects (30% women, M = 33.4 yr old) during the 6° head-down tilt bed rest study AGBRESA during 20 training sessions with the 6df learning program for spacecraft docking.RESULTS: Subjects' gaze was directed most frequently and longest to the vizor (185 dwells and 22,355 ms per task) followed by the two instrument displays (together 75 dwells and 4048 ms per task). We observed a significant positive relationship between number and duration of visual checks of speed and distance to the docking point and the accuracy of the docking maneuver.DISCUSSION: In conclusion, eye tracking provides valuable information related to docking accuracy that might prospectively offer the opportunity to improve docking training effectiveness.Piechowski S, Johannes B, Pustowalow W, Arz M, Mulder E, Jordan J, Wolf OT, Rittweger J. Visual attention relates to operator performance in spacecraft docking training. Aerosp Med Hum Perform. 2022; 93(6):480-486.

The Effects of 30 Minutes of Artificial Gravity on Cognitive and Sensorimotor Performance in a Spaceflight Analog Environment.

The altered vestibular signaling and somatosensory unloading of microgravity result in sensory reweighting and adaptation to conflicting sensory inputs. Aftereffects of these adaptive changes are evident postflight as impairments in behaviors such as balance and gait. Microgravity also induces fluid shifts toward the head and an upward shift of the brain within the skull; these changes are well-replicated in strict head-down tilt bed rest (HDBR), a spaceflight analog environment. Artificial gravity (AG) is a potential countermeasure to mitigate these effects of microgravity. A previous study demonstrated that intermittent (six, 5-mins bouts per day) daily AG sessions were more efficacious at counteracting orthostatic intolerance in a 5 day HDBR study than continuous daily AG. Here we examined whether intermittent daily AG was also more effective than continuous dosing for mitigating brain and behavioral changes in response to 60 days of HDBR. Participants (n = 24) were split evenly between three groups. The first received 30 mins of continuous AG daily (cAG). The second received 30 mins of intermittent AG daily (6 bouts of 5 mins; iAG). The third received no AG (Ctrl). We collected a broad range of sensorimotor, cognitive, and brain structural and functional assessments before, during, and after the 60 days of HDBR. We observed no significant differences between the three groups in terms of HDBR-associated changes in cognition, balance, and functional mobility. Interestingly, the intermittent AG group reported less severe motion sickness symptoms than the continuous group during centrifugation; iAG motion sickness levels were not elevated above those of controls who did not undergo AG. They also had a shorter duration of post-AG illusory motion than cAG. Moreover, the two AG groups performed the paced auditory serial addition test weekly while undergoing AG; their performance was more accurate than that of controls, who performed the test while in HDBR. Although AG did not counteract HDBR-induced gait and balance declines, iAG did not cause motion sickness and was associated with better self-motion perception during AG ramp-down. Additionally, both AG groups had superior cognitive performance while undergoing AG relative to controls; this may reflect attention or motivation differences between the groups.

Head-Down-Tilt Bed Rest With Elevated CO2: Effects of a Pilot Spaceflight Analog on Neural Function and Performance During a Cognitive-Motor Dual Task.

Spaceflight has widespread effects on human performance, including on the ability to dual task. Here, we examine how a spaceflight analog comprising 30 days of head-down-tilt bed rest (HDBR) combined with 0.5% ambient CO2 (HDBR + CO2) influences performance and functional activity of the brain during single and dual tasking of a cognitive and a motor task. The addition of CO2 to HDBR is thought to better mimic the conditions aboard the International Space Station. Participants completed three tasks: (1) COUNT: counting the number of times an oddball stimulus was presented among distractors; (2) TAP: tapping one of two buttons in response to a visual cue; and (3) DUAL: performing both tasks concurrently. Eleven participants (six males) underwent functional MRI (fMRI) while performing these tasks at six time points: twice before HDBR + CO2, twice during HDBR + CO2, and twice after HDBR + CO2. Behavioral measures included reaction time, standard error of reaction time, and tapping accuracy during the TAP and DUAL tasks, and the dual task cost (DTCost) of each of these measures. We also quantified DTCost of fMRI brain activation. In our previous HDBR study of 13 participants (with atmospheric CO2), subjects experienced TAP accuracy improvements during bed rest, whereas TAP accuracy declined while in the current study of HDBR + CO2. In the HDBR + CO2 subjects, we identified a region in the superior frontal gyrus that showed decreased DTCost of brain activation while in HDBR + CO2, and recovered back to baseline levels before the completion of bed rest. Compared to HDBR alone, we found different patterns of brain activation change with HDBR + CO2. HDBR + CO2 subjects had increased DTCost in the middle temporal gyrus whereas HDBR subjects had decreased DTCost in the same area. Five of the HDBR + CO2 subjects developed signs of spaceflight-associated neuro-ocular syndrome (SANS). These subjects exhibited lower baseline dual task activation and higher slopes of change during HDBR + CO2 than subjects with no signs of SANS. Collectively, this pilot study provides insight into the additional and/or interactive effects of CO2 levels during HDBR, and information regarding the impacts of this spaceflight analog environment on the neural correlates of dual tasking.

Long-Term Bed Rest Delays the Circadian Phase of Core Body Temperature.

Spaceflight can be associated with sleep loss and circadian misalignment as a result of non-24 h light-dark cycles, operational shifts in work/rest cycles, high workload under pressure, and psychological factors. Head-down tilt bed rest (HDBR) is an established model to mimic some of the physiological and psychological adaptions observed in spaceflight. Data on the effects of HDBR on circadian rhythms are scarce. To address this gap, we analyzed the change in the circadian rhythm of core body temperature (CBT) in two 60-day HDBR studies sponsored by the European Space Agency [n = 13 men, age: 31.1 ± 8.2 years (M ± SD)]. CBT was recorded for 36 h using a non-invasive and validated dual-sensor heatflux technology during the 3rd and the 8th week of HDBR. Bed rest induced a significant phase delay from the 3rd to the 8th week of HDBR (16.23 vs. 16.68 h, p = 0.005, g = 0.85) irrespective of the study site (p = 0.416, g = −0.46), corresponding to an average phase delay of about 0.9 min per day of HDBR. In conclusion, long-term bed rest weakens the entrainment of the circadian system to the 24-h day. We attribute this effect to the immobilization and reduced physical activity levels associated with HDBR. Given the critical role of diurnal rhythms for various physiological functions and behavior, our findings highlight the importance of monitoring circadian rhythms in circumstances in which gravity or physical activity levels are altered.

Head-Down Tilt Bed Rest Studies as a Terrestrial Analog for Spaceflight Associated Neuro-Ocular Syndrome.

Astronauts who undergo prolonged periods of spaceflight may develop a unique constellation of neuro-ocular findings termed Spaceflight Associated Neuro-Ocular Syndrome (SANS). SANS is a disorder that is unique to spaceflight and has no terrestrial equivalent. The prevalence of SANS increases with increasing spaceflight duration and although there have been residual, structural, ocular changes noted, no irreversible or permanent visual loss has occurred after SANS, with the longest spaceflight to date being 14 months. These microgravity-induced findings are being actively investigated by the United States' National Aeronautics Space Administration (NASA) and SANS is a potential obstacle to future longer duration, manned, deep space flight missions. The pathophysiology of SANS remains incompletely understood but continues to be a subject of intense study by NASA and others. The study of SANS is of course partially limited by the small sample size of humans undergoing spaceflight. Therefore, identifying a terrestrial experimental model of SANS is imperative to facilitate its study and for testing of preventative measures and treatments. Head-down tilt bed rest (HDTBR) on Earth has emerged as one promising possibility. In this paper, we review the HDTBR as an analog for SANS pathogenesis; the clinical and imaging overlap between SANS and HDTBR studies; and potential SANS countermeasures that have been or could be tested with HDTBR.

Continuous and Intermittent Artificial Gravity as a Countermeasure to the Cognitive Effects of 60 Days of Head-Down Tilt Bed Rest.

Environmental and psychological stressors can adversely affect astronaut cognitive performance in space. This study used a 6° head-down tilt bed rest (HDBR) paradigm to simulate some of the physiologic changes induced by microgravity. Twenty-four participants (mean ± SD age 33.3 ± 9.2 years, N = 16 men) spent 60 consecutive days in strict HDBR. They were studied in three groups of eight subjects each. One group served as Control, whereas the other two groups received either a continuous or intermittent artificial gravity (AG) countermeasure of 30 min centrifugation daily (1 g acceleration at the center of mass and 2 g at the feet). Participants performed all 10 tests of NASA’s Cognition battery and a brief alertness and mood survey repeatedly before, during, and after the HDBR period. Test scores were adjusted for practice and stimulus set difficulty effects. A modest but statistically significant slowing across a range of cognitive domains was found in all three groups during HDBR compared to baseline, most consistently for sensorimotor speed, whereas accuracy was unaffected. These changes were observed early during HDBR and did not further worsen or improve with increasing time in HDBR, except for emotion recognition performance. With increasing time spent in HDBR, participants required longer time to decide which facial emotion was expressed. They were also more likely to select categories with negative valence over categories with neutral or positive valence. Except for workload, which was rated lower in the Control group, continuous or intermittent AG did not modify the effect of HDBR on cognitive performance or subjective responses. Participants expressed several negative survey responses during HDBR relative to baseline, and some of the responses further deteriorated during recovery, which highlights the importance of adequate medical and psychological support during extended duration HDBR studies. In conclusion, 60 days of HDBR were associated with moderate cognitive slowing and changes in emotion recognition performance, but these effects were not mitigated by either continuous or intermittent exposure to AG for 30 min daily.

Bone Marrow Reconversion With Reambulation: A Prospective Clinical Trial.

In a prospective clinical trial, 20 healthy men participated in a 60-day, 6-degree head-down tilt bed rest study. Serial 3-T magnetic resonance (MR) imaging measures of the lumbar spine were performed at baseline, after 57 days of bed rest, and at 30, 360, and 720 days of reambulation (100 MR imaging scans). Proton density with and without fat saturation, 2-point Dixon, and single-voxel MR spectroscopy techniques were used to assess bone marrow composition (300 measures). Erythropoiesis was measured using hematocrit, reticulocyte, and ferritin. Also, participants randomly received either a nutritional intervention composed of polyphenols, omega-3, vitamin E, and selenium or a normal diet. Thirty days of reambulation after 60 days of bed rest caused a marked decrease of the mean lumbar vertebral fat fraction (VFF) (-9.2 ± 1.6 percentage points, -8.0 ± 1.3 percentage points, and -12.7 ± 1.2 percentage points compared with baseline using proton density, Dixon, MR spectroscopy, respectively; all 3, P < 0.05). Reambulation also decreased the fat saturation index (-5.3 ± 1.1 percentage points compared with baseline; P < 0.05). These coincided with lower hematocrit and ferritin and with increased reticulocytes at reambulation day 13 compared with baseline (all 3, P < 0.05). After 57 days of bed rest, the VFF was unchanged from baseline (all 3 MR techniques, P > 0.05); reambulation for 2 years returned the lumbar VFF to baseline values. This longitudinal trial established that 30 days of reambulation after 60 days of bed rest constituted a powerful stimulus for bone marrow reconversion. In this model, the enhanced erythropoiesis coupled with preferential consumption of fatty acids from regulated marrow adipose tissue to supply energy for erythropoiesis and bone anabolism may explain the lumbar vertebrae reconversion. These results will help interpreting bone marrow signal in ambulatory patients after long periods of bed rest.

Neural Working Memory Changes During a Spaceflight Analog With Elevated Carbon Dioxide: A Pilot Study.

Spaceflight missions to the International Space Station (ISS) expose astronauts to microgravity, radiation, isolation, and elevated carbon dioxide (CO2), among other factors. Head down tilt bed rest (HDBR) is an Earth-based analog for spaceflight used to study body unloading, fluid shifts, and other factors unrelated to gravitational changes. While in space, astronauts need to use mental rotation strategies to facilitate their adaptation to the ISS environment. Therefore, spatial working memory is essential for crewmember performance. Although the effects of HDBR on spatial working memory have recently been studied, the results are still inconclusive. Here, we expand upon past work and examine the effects of HDBR with elevated CO2 (HDBR + CO2) on brain activation patterns during spatial working memory performance. In addition, we compare brain activation between 30 days of HDBR + CO2 and 70 days of HDBR to test the isolated effect of CO2. Eleven subjects (6 males, 5 females; mean age = 34 ± 8 years) underwent six functional magnetic resonance imaging (fMRI) sessions pre-, during, and post-HDBR + CO2. During the HDBR + CO2 intervention, we observed decreasing activation in the right middle frontal gyrus and left regions of the cerebellum, followed by post-intervention recovery. We detected several correlations between brain and behavioral slopes of change with the HDBR + CO2 intervention. For example, greater increases in activation in frontal, temporal and parietal regions were associated with larger spatial working memory improvements. Comparing the HDBR + CO2 group to data from our previous 70-day HDBR study, we found greater decreases in activation in the right hippocampus and left inferior temporal gyrus for the HDBR + CO2 group over the course of the intervention. Together, these findings increase our understanding of the neural mechanisms of HDBR, elevated levels of CO2 and spaceflight-related changes in spatial working memory performance.

MTRR 66 and SHMT1 1420 Variants Are Associated with Optic Disc Edema During 30-d Strict Head-down Tilt Bed Rest and CO2 Exposure (P24-036-19)

ObjectivesSome astronauts returning from missions to the International Space Station (ISS) have developed ophthalmic structural changes, including optic disc edema. Incidence of optic disc edema among astronauts has been linked to one-carbon pathway genetic variants and B-vitamin status. A recent 30-d 6-degree head-down tilt bed rest study in which all subjects were exposed to 0.5% CO2 documented the occurrence of optic disc edema in 5 of 11 subjects. The objective of this study was to determine the effect of one-carbon pathway genetics and B-vitamin status on the incidence of optic disc edema in the bed rest subjects. MethodsThe study was conducted at the: envihab facility at the German Aerospace Center in Cologne, Germany. Subjects (6 M/5F) were healthy volunteers, 33 ± 8 y (mean ± SD), having a mean BMI of 23.4 ± 2.2 kg/m2. After a 14-d ambulatory phase in a standard environment (i.e., not hypercapnic), a blood sample was collected to assess vitamin status and single nucleotide polymorphisms (SNPs) associated with one carbon metabolism. We focused on two SNPs (rs1801394, MTRR 66 and rs1979227, SHMT1 1420) based on our earlier findings in astronauts (Zwart et al., FASEB J, 30:141–8, 2016). The 30-d bed rest began after collection of this sample. Optical coherence tomography images were collected before, during (days 15 and 30), and after (6 and 13 days after reambulation) bed rest, and the change in total retinal thickness (TRT) at various distances from Bruch’s membrane opening (BMO) was evaluated against the number of MTRR A66G G alleles and SHMT1 C1420T C alleles. ResultsThe change in TRT at various distances from the BMO in bed rest subjects exposed to mild hypercapnia and headward fluid shifts was strongly associated with the number of MTRR 66 G and SHMT1 1420 C alleles. ConclusionsThis finding may increase understanding of mechanisms involved in optic disc edema in astronauts and patients on Earth, and hence lead to development of targeted countermeasures. Funding SourcesThis work was funded by the NASA Human Health Countermeasures Element of the NASA Human Research Program.

Change of cortical foot activation following 70 days of head-down bed rest.

Head-down tilt bed rest (HDBR) has been used as a spaceflight analog to study some of the effects of microgravity on human physiology, cognition, and sensorimotor functions. Previous studies have reported declines in balance control and functional mobility after spaceflight and HDBR. In this study we investigated how the brain activation for foot movement changed with HDBR. Eighteen healthy men participated in the current HDBR study. They were in a 6° head-down tilt position continuously for 70 days. Functional MRI scans were acquired to estimate brain activation for foot movement before, during, and after HDBR. Another 11 healthy men who did not undergo HDBR participated as control subjects and were scanned at four time points. In the HDBR subjects, the cerebellum, fusiform gyrus, hippocampus, and middle occipital gyrus exhibited HDBR-related increases in activation for foot tapping, whereas no HDBR-associated activation decreases were found. For the control subjects, activation for foot tapping decreased across sessions in a couple of cerebellar regions, whereas no activation increase with session was found. Furthermore, we observed that less HDBR-related decline in functional mobility and balance control was associated with greater pre-to-post HDBR increases in brain activation for foot movement in several cerebral and cerebellar regions. Our results suggest that more neural control is needed for foot movement as a result of HDBR. NEW & NOTEWORTHY Long-duration head-down bed rest serves as a spaceflight analog research environment. We show that brain activity in the cerebellum and visual areas during foot movement increases from pre- to post-bed rest and then shows subsequent recovery. Greater increases were seen for individuals who exhibited less decline in functional mobility and balance control, suggestive of adaptive changes in neural control with long-duration bed rest.

Long-duration bed rest modifies sympathetic neural recruitment strategies in male and female participants.

To understand the impact of physical deconditioning with head-down tilt bed rest (HDBR) on the malleability of sympathetic discharge patterns, we studied 1) baseline integrated muscle sympathetic nerve activity (MSNA; microneurography) from 13 female participants in the WISE-2005 60-day HDBR study (retrospective analysis), 2) integrated MSNA and multiunit action potential (AP) analysis in 13 male participants performed on data collected at baseline and during physiological stress imposed by end-inspiratory apnea in a new 60-day HDBR study, and 3) a repeatability study (control; n = 6, retrospective analysis, 4 wk between tests). Neither baseline integrated burst frequency nor incidence were altered with HDBR (both P > 0.35). However, baseline integrated burst latency increased in both HDBR studies (male: 1.35 ± 0.02 to 1.39 ± 0.02 s, P < 0.01; female: 1.23 ± 0.02 to 1.29 ± 0.02 s, P < 0.01), whereas controls exhibited no change across two visits (1.25 ± 0.02 to 1.25 ± 0.02 s, group-by-time interaction, P = 0.02). With the exception of increased AP latency ( P = 0.03), male baseline AP data did not change with HDBR (all P > 0.19). The change in AP frequency on going from baseline to apnea (∆94 ± 25 to ∆317 ± 55 AP/min, P < 0.01) and the number of active sympathetic clusters per burst (∆0 ± 0.2 to ∆1 ± 0.2 clusters/burst, P = 0.02) were greater post- compared with pre-HDBR. The change in total clusters with apnea was ∆0 ± 0.5 clusters pre- and ∆2 ± 0.7 clusters post-HDBR ( P = 0.07). These data indicate that 60-day HDBR modified discharge characteristics in baseline burst latency and sympathetic neural recruitment during apneic stress. NEW & NOTEWORTHY Long-duration bed rest did not modify baseline sympathetic burst frequency in male and female participants, but examination of additional features of the multiunit signal provided novel evidence to suggest augmented synaptic delays or processing times at baseline for all sympathetic action potentials. Furthermore, long-duration bed rest increased reflex-sympathetic arousal to apneic stress in male participants primarily by mechanisms involving an augmented firing rate of action potential clusters active at baseline.

Exercise as potential countermeasure for the effects of 70 days of bed rest on cognitive and sensorimotor performance.

Background: Spaceflight has been associated with changes in gait and balance; it is unclear whether it affects cognition. Head down tilt bed rest (HDBR) is a microgravity analog that mimics cephalad fluid shifts and body unloading. In consideration of astronaut’s health and mission success, we investigated the effects of HDBR on cognition and sensorimotor function. Furthermore, we investigated if exercise mitigates any cognitive and sensorimotor sequelae of spaceflight.Method: We conducted a 70-day six-degree HDBR study in 10 male subjects who were randomly assigned to a HDBR supine exercise or a HDBR control group. Cognitive measures (i.e., processing speed, manual dexterity, psychomotor speed, visual dependency, and 2D and 3D mental rotation) and sensorimotor performance (functional mobility (FMT) and balance performance) were collected at 12 and 8 days pre-HDBR, at 7, 50, and 70 days in HDBR, and at 8 and 12 days post-HDBR. Exercise comprised resistance training, and continuous and high-intensity interval aerobic exercise. We also repeatedly assessed an outside-of-bed rest control group to examine metric stability.Results: Small practice effects were observed in the control group for some tasks; these were taken into account when analyzing effects of HDBR. No significant effects of HDBR on cognition were observed, although visual dependency during HDBR remained stable in HDBR controls whereas it decreased in HDBR exercise subjects. Furthermore, HDBR was associated with loss of FMT and standing balance performance, which were almost fully recovered 12 days post-HDBR. Aerobic and resistance exercise partially mitigated the effects of HDBR on FMT and accelerated the recovery time course post-HDBR.Discussion: HDBR did not significantly affect cognitive performance but did adversely affect FMT and standing balance performance. Exercise had some protective effects on the deterioration and recovery of FMT.

Energy expenditure and balance during spaceflight on the space shuttle.

The objectives of this study were as follows: 1) to measure human energy expenditure (EE) during spaceflight on a shuttle mission by using the doubly labeled water (DLW) method; 2) to determine whether the astronauts were in negative energy balance during spaceflight; 3) to use the comparison of change in body fat as measured by the intake DLW EE, 18O dilution, and dual energy X-ray absorptiometry (DEXA) to validate the DLW method for spaceflight; and 4) to compare EE during spaceflight against that found with bed rest. Two experiments were conducted: a flight experiment (n = 4) on the 16-day 1996 life and microgravity sciences shuttle mission and a 6 degrees head-down tilt bed rest study with controlled dietary intake (n = 8). The bed rest study was designed to simulate the flight experiment and included exercise. Two EE determinations were done before flight (bed rest), during flight (bed rest), and after flight (recovery). Energy intake and N balance were monitored for the entire period. Results were that body weight, water, fat, and energy balance were unchanged with bed rest. For the flight experiment, decreases in weight (2.6 +/- 0.4 kg, P < 0.05) and N retention (-2. 37 +/- 0.45 g N/day, P < 0.05) were found. Dietary intake for the four astronauts was reduced in flight (3,025 +/- 180 vs. 1,943 +/- 179 kcal/day, P < 0.05). EE in flight was 3,320 +/- 155 kcal/day, resulting in a negative energy balance of 1,355 +/- 80 kcal/day (-15. 7 +/- 1.0 kcal. kg-1. day-1, P < 0.05). This corresponded to a loss of 2.1 +/- 0.4 kg body fat, which was within experimental error of the fat loss determined by 18O dilution (-1.4 +/- 0.5 kg) and DEXA (-2.4 +/- 0.4 kg). All three methods showed no change in body fat with bed rest. In conclusion, 1) the DLW method for measuring EE during spaceflight is valid, 2) the astronauts were in severe negative energy balance and oxidized body fat, and 3) in-flight energy (E) requirements can be predicted from the equation: E = 1.40 x resting metabolic rate + exercise.