Comprehensive assessment of physiological responses in women during the ESA dry immersion VIVALDI microgravity simulation.

Astronauts in microgravity experience multi-system deconditioning, impacting their inflight efficiency and inducing dysfunctions upon return to Earth gravity. To fill the sex gap of knowledge in the health impact of spaceflights, we simulate microgravity with a 5-day dry immersion in 18 healthy women (ClinicalTrials.gov Identifier: NCT05043974). Here we show that dry immersion rapidly induces a sedentarily-like metabolism shift mimicking the beginning of a metabolic syndrome with a drop in glucose tolerance, an increase in the atherogenic index of plasma, and an impaired lipid profile. Bone remodeling markers suggest a decreased bone formation coupled with an increased bone resorption. Fluid shifts and muscular unloading participate to a marked cardiovascular and sensorimotor deconditioning with decreased orthostatic tolerance, aerobic capacity, and postural balance. Collected datasets provide a comprehensive multi-systemic assessment of dry immersion effects in women and pave the way for future sex-based evaluations of countermeasures.

Integrated Biophysical and Biochemical Signals Augment Megakaryopoiesis and Thrombopoiesis in a Three-Dimensional Rotary Culture System.

Platelet transfusion has been widely used in patients undergoing chemotherapy or radiotherapy; however, the shortage of the platelet supply limits the care of patients. Although derivation of clinical-scale platelets in vitro could provide a new source for transfusion, the devices and procedures for deriving scalable platelets for clinical applications have not been established. In the present study, we found that a rotary cell culture system (RCCS) can potentiate megakaryopoiesis and significantly improve the efficiency of platelet generation. When used with chemical compounds and growth factors identified via small-scale screening, the RCCS improved platelet generation efficiency by as much as ∼3.7-fold compared with static conditions. Shear force, simulated microgravity, and better diffusion of nutrients and oxygen from the RCCS, altogether, might account for the improved efficient platelet generation. The cost-effective and highly controllable strategy and methodology represent an important step toward large-scale platelet production for future biomedical and clinical applications. Significance: Platelet transfusion has been widely used in patients undergoing chemotherapy or radiotherapy; however, the shortage of platelet supply limits the care of patients. Thus, derivation of clinical-scale platelets in vitro would provide a new source for transfusion. The present study evaluated a rotary suspension cell culture system that was able to potentiate megakaryopoiesis and significantly improved the efficiency of platelet generation. When used with chemical compounds and growth factors identified via small-scale screening, the three-dimensional system improved platelet generation efficiency compared with the static condition. The three-dimensional device and the strategy developed in the present study should markedly improve the generation of large-scale platelets for use in future biomedical and clinical settings.

[HEART RHYTHM VARIABILITY ANALYSIS AND ASSESSMENT OF THE SPINAL PAIN SYNDROME DURING DRY IMMERSION].

The spinal pain syndrome appears in cosmonauts on both short and long-duration missions. This untoward factor may affect body systems functioning and complicate the successful accomplishment of space mission. Purpose of the investigation was to examine the lumbar spine and to elucidate whether its condition relates to the spinal pain development and changes in heart rate variability (HRV) in the microgravity environment. The experiment was conducted in dry immersion as a method of microgravity effects simulation. It was shown that in dry immersion locomotion reproduces the patterns peculiar for significant gravitational unloading. Spinal pain intensity, angles and heights of the lumbar intervertebral discs and HRV were measured in 19 selected volunteers. During the experiment, all the volunteers developed pains in the back that abated gradually. Pain dependence on the height of intervertebral discs and cardiac regulatory mechanisms were investigated.

Simulated microgravity: critical review on the use of random positioning machines for mammalian cell culture.

Random Positioning Machines (RPMs) have been used since many years as a ground-based model to simulate microgravity. In this review we discuss several aspects of the RPM. Recent technological development has expanded the operative range of the RPM substantially. New possibilities of live cell imaging and partial gravity simulations, for example, are of particular interest. For obtaining valuable and reliable results from RPM experiments, the appropriate use of the RPM is of utmost importance. The simulation of microgravity requires that the RPM's rotation is faster than the biological process under study, but not so fast that undesired side effects appear. It remains a legitimate question, however, whether the RPM can accurately and reliably simulate microgravity conditions comparable to real microgravity in space. We attempt to answer this question by mathematically analyzing the forces working on the samples while they are mounted on the operating RPM and by comparing data obtained under real microgravity in space and simulated microgravity on the RPM. In conclusion and after taking the mentioned constraints into consideration, we are convinced that simulated microgravity experiments on the RPM are a valid alternative for conducting examinations on the influence of the force of gravity in a fast and straightforward approach.

An assessment of the long-term effects of simulated microgravity on cranial neural crest cells in zebrafish embryos with a focus on the adult skeleton.

It is becoming increasingly important to address the long-term effects of exposure to simulated microgravity as the potential for space tourism and life in space become prominent topics amongst the World's governments. There are several studies examining the effects of exposure to simulated microgravity on various developmental systems and in various organisms; however, few examine the effects beyond the juvenile stages. In this study, we expose zebrafish embryos to simulated microgravity starting at key stages associated with cranial neural crest cell migration. We then analyzed the skeletons of adult fish. Gross observations and morphometric analyses show that exposure to simulated microgravity results in stunted growth, reduced ossification and severe distortion of some skeletal elements. Additionally, we investigated the effects on the juvenile skull and body pigmentation. This study determines for the first time the long-term effects of embryonic exposure to simulated microgravity on the developing skull and highlights the importance of studies investigating the effects of altered gravitational forces.

The role of echocardiography in the assessment of cardiac function in weightlessness-Our experience during parabolic flights.

Parabolic flight (PF) elicits changes in hydrostatic pressure gradients, resulting in increase (at 0Gz) or decrease (at 1.8Gz) in cardiac preload. The magnitude of these changes on left ventricular (LV) and atrial (LA) volumes, as well as on myocardial velocities, strain and strain rates, is largely unknown. Using real-time 3D (RT3DE) and Doppler tissue echocardiographic imaging (DTI) during PF in normal subjects in standing position, we showed that both LV and LA volumes were decreased at 1.8Gz and increased at 0Gz by about 20% and 40%, respectively. Previous 2D or M-mode studies underestimated such changes. Also, preload dependence was confirmed for systolic and diastolic velocities, and peak systolic strain, while strain rates were preload independent, probably reflecting intrinsic myocardial properties. Low body negative pressure at -50mmHg applied during 0Gz was effective in restoring 1Gz levels. RT3DE and DTI during PF are feasible, allowing the evaluation of the cardiac function under different loading conditions.

Skinfold thickness versus isotope dilution for body fat assessment during simulated microgravity: results from three bed-rest campaigns in men and women with and without countermeasures.

Because body composition is altered during head-down bed rest (HDBR), body mass can not be used as an index of energy balance. Consequently diet allowances should not be based on body mass evolution but on fat mass changes. Though criticized, skinfold thickness (ST) is the costless, easiest and fastest method to use for such an objective. The aim of this study was to compare the percentage of body fat (%BF) estimated by ST with the isotope dilution of H2 18O. We compiled data from three HDBR campaigns, one on women (n=8) in November 1998 and two on the same men (n=8) in December 1997 (without countermeasure) and January 1998 (with thigh-cuffs countermeasure), according to a crossover design. Body composition was assessed before and after 6 days of HDBR. %BF was derived from the biceps, triceps, sub-scapular and sup-iliac ST according to Durnin and Wormersly (1974). Fat-free mass was measured on the same day by H2 18O dilution and fat mass was calculated by the difference with body mass and expressed as a percentage. Based on precision tests, the minimum measurable change by ST was 1.1%BF for single measurement point. Both intercepts (F (4,30)=0.89, P=0.45) and slopes (F (4,30)=0.74; P=0.57) of the ST versus dilution relationships were not affected by the periods (December vs January), experimental conditions (control vs HDBR vs HDBR + thigh cuffs) or sex allowing the derivation of a common relationship %BF(st)=0.94 x %BF(dil) (F (1,47)=97.9, P<0.0001; non-significant intercept excluded) with a bias between methods of -1.7+/-2.0 %BF (95% CI: -5.8, 2.4 %BF). ST can be used to measure %BF during HDBR provided great care is placed on training and changes are higher than 1.1 %BF. If the method can be applied for in-flight energy balance monitoring given the high observed energy deficit, a tight monitoring of the individual nutritional status as needed during simulation appears, however, dubious based on this solely method.

Assessment of simulated surgical skills in parabolic microgravity.

BACKGROUND: During spaceflight crew health is paramount in the success of flight missions. The delivery of healthcare during flight requires crew readiness for medical and surgical response. METHODS: There were 20 participants who were evaluated for accurate performance of 4 basic laparoscopic surgical skills (clip applying, cutting, grasping, and suturing) during parabolic weightlessness using an inanimate workstation aboard the NASA KC-135 aircraft. RESULTS: Data indicate that motor skill performance decreased within the parabolic microgravity flight environment. Performance in parabolic microgravity flight included futile effort with an increase in number of tasks attempted and a decrease in tasks completed successfully. CONCLUSIONS: There is a decreased frequency of accurate task completion in parabolic microgravity flight, but it is not an obstacle to implementation of effective training for providing in-flight medical care. The data reveal that individuals perform basic laparoscopic surgical simulation with greater effort in microgravity following simulation training.

Independent metabolic costs of supporting body weight and accelerating body mass during walking.

The metabolic cost of walking is determined by many mechanical tasks, but the individual contribution of each task remains unclear. We hypothesized that the force generated to support body weight and the work performed to redirect and accelerate body mass each individually incur a significant metabolic cost during normal walking. To test our hypothesis, we measured changes in metabolic rate in response to combinations of simulated reduced gravity and added loading. We found that reducing body weight by simulating reduced gravity modestly decreased net metabolic rate. By calculating the metabolic cost per Newton of reduced body weight, we deduced that generating force to support body weight comprises approximately 28% of the metabolic cost of normal walking. Similar to previous loading studies, we found that adding both weight and mass increased net metabolic rate in more than direct proportion to load. However, when we added mass alone by using a combination of simulated reduced gravity and added load, net metabolic rate increased about one-half as much as when we added both weight and mass. By calculating the cost per kilogram of added mass, we deduced that the work performed on the center of mass comprises approximately 45% of the metabolic cost of normal walking. Our findings support the hypothesis that force and work each incur a significant metabolic cost. Specifically, the cost of performing work to redirect and accelerate the center of mass is almost twice as great as the cost of generating force to support body weight.

Focused Assessment with Sonography for Trauma in weightlessness: a feasibility study.

BACKGROUND: The Focused Assessment with Sonography for Trauma (FAST) examines for fluid in gravitationally dependent regions. There is no prior experience with this technique in weightlessness, such as on the International Space Station, where sonography is currently the only diagnostic imaging tool. STUDY DESIGN: A ground-based (1 g) porcine model for sonography was developed. We examined both the feasibility and the comparative performance of the FAST examination in parabolic flight. Sonographic detection and fluid behavior were evaluated in four animals during alternating weightlessness (0 g) and hypergravity (1.8 g) periods. During flight, boluses of fluid were incrementally introduced into the peritoneal cavity. Standardized sonographic windows were recorded. Postflight, the video recordings were divided into 169 20-second segments for subsequent interpretation by 12 blinded ultrasonography experts. Reviewers first decided whether a video segment was of sufficient diagnostic quality to analyze (determinate). Determinate segments were then analyzed as containing or not containing fluid. A probit regression model compared the probability of a positive fluid diagnosis to actual fluid levels (0 to 500 mL) under both 0-g and 1.8-g conditions. RESULTS: The in-flight sonographers found real-time scanning and interpretation technically similar to that of terrestrial conditions, as long as restraint was maintained. On blinded review, 80% of the recorded ultrasound segments were considered determinate. The best sensitivity for diagnosis in 0 g was found to be from the subhepatic space, with probability of a positive fluid diagnosis ranging from 9% (no fluid) to 51% (500 mL fluid). CONCLUSIONS: The FAST examination is technically feasible in weightlessness, and merits operational consideration for clinical contingencies in space.

Tissue engineering in space.

The purpose of this paper is to present the status of that part of the [Microgravity Application Program] project related to the study of cartilage formation from pig chondrocytes. The work carried out so far followed two lines: (i) chondrocytes were incubated for up to three weeks in the RPM; (ii) a module developed for in-vitro cartilage formation will be tested in a sounding rocket flight (MASER 9, November 2001).

Simulated weightlessness alters the nycthemeral distribution of energy expenditure in rats.

The energy metabolism adaptations to simulated weightlessness in rats by hindlimb tail suspension are unknown. 12 male rats were assigned to 7 days of isolation, 7 days of habituation to the suspension device, 10 days of simulated weightlessness, and 3 days of recovery. The 24-hour energy expenditure was measured by continuous indirect calorimetry. We calculated the 12-hour energy expenditure during the active (night) and inactive (day) periods, the minimal observed metabolic rates with the day values taken as an index of the basal metabolic rate, and the non-basal energy expenditure representing the cost of physical activity plus the diet-induced thermogenesis. Suspension did not change the mean 24-hour energy expenditure (360.8+/-15.3 J min(-1) kg(-0.67)), but reduced the night/day difference by 64 % (P<0.05) through a concomitant drop in night-energy expenditure and increase in day values. The difference between night and day minimal metabolic rates was reduced by 81 % (P<0.05), and the transient rise in day values suggests an early and moderate basal metabolic rate increase (9 %). An overall 19 % reduction in non-basal energy expenditure was observed during simulated weightlessness (P<0.05), which was mainly attributable to a reduction in the cost of physical activity. 3 days of recovery restored the night/day differences but increased the 24-hour energy expenditure by 10 % (P<0.05). In conclusion, hindlimb tail suspension in rats did not alter the 24-hour energy expenditure, but it transiently increased the basal metabolic rate, and altered both the energy expended on physical activity and the nycthemeral distribution of motor activity. These data suggest that the circadian rhythms of energy expenditure are affected during simulated weightlessness.

Intracranial pressure in microgravity conditions: non-invasive assessment by ophthalmodynamometry.

BACKGROUND: As well known from former manned spaceflight experiments (German D2-Mission/German D1-Mission 1985/German-Russian MIR-Mission 1 992/German-D2-Mission 1993), fluid shift after entry into microgravity leads to a rapid increase in pressure and volume within the upper compartments of the human body. This has been proven by precise measurements with automatic selftonometers for intraocular pressure. HYPOTHESIS: There is little doubt, that a very similar--even more, marked--increase of intracranial pressure happens soon after entry into microgravity. This may be the cause for some of the reported hormonal and even neurological changes in metabolism. There is no non-invasive method to assess these important increases in pressure. METHODS: Ophthalmodynamometry in general allows for rather precise estimation of intracranial BP, but so far the method was too complicated for routine application, specifically in spaceflight conditions. Therefore, using the microprocessor controlled technology of our automatic selftonometer we have designed a very precise automatic instrument which can be applied by the astronaut/kosmonaut. The measurement takes only a few seconds. CONCLUSIONS: This easily applied, non-invasive method would allow for completely new insights into these important changes and explain some of the clinical consequences noted so far.

[Echocardiographic assessment of left ventricular structure and function after simulated weightlessness in rats].

Objective. To investigate whether the changes in rat after simulated weightlessness are similar to those in astronaut after flight. Methods. The effects of 4 wk tail-suspension on left ventricular structure and function in rats were examined by echocardiography. Results. After 4 wk of simulated weightlessness, the thickness of both the anterior and posterior wall in left ventricle (LV) showed a general trend of decrease, but these changes were not statistically significant; the end-systolic and end-diastolic internal dimensions (ESD and EDD respectively) of LV decreased significantly; and the end-systolic volume, end-diastolic volume and stroke volume (ESV, EDV and SV respectively) were all reduced; so did the relevant indices of them. There were no significant differences in ejection fraction (EF) and fractional shortening (FS) between the tail-suspended and control groups. The left ventricular mass (LVM) and its index (LVMI) were decreased. The peak velocities of blood flow of aorta, pulmonary artery and mitral valve didn't show any significant change after simulated weightlessness. Conclusion. Medium-term simulated weightlessness may lead to a significant decrease in left ventricular internal dimension, ventricular volume, and mass, and a trend of decrease in mean left ventricular wall thickness. These changes in rats are similar to those observed in astronauts postflight.

The NASA Performance Assessment Workstation: cognitive performance during head-down bed rest.

The NASA Performance Assessment Workstation was used to assess cognitive performance changes in eight males subjected to seventeen days of 6 degrees head-down bed rest. PAWS uses six performance tasks to assess directed and divided attention, spatial, mathematical, and memory skills, and tracking ability. Subjective scales assess overall fatigue and mood state. Subjects completed training trials, practice trials, bed rest trials, and recovery trials. The last eight practice trials and all bed rest trials were performed with subjects lying face-down on a gurney. In general, there was no apparent cumulative effect of bed rest. Following a short period of performance stabilization, a slight but steady trend of performance improvement was observed across all trials. For most tasks, this trend of performance improvement was enhanced during recovery. No statistically significant differences in performance were observed when comparing bed rest with the control period. Additionally, fatigue scores showed little change across all periods.

Physiological [correction of Physilogical] cost of standard physical load after 7-day dry immersion.

NASA: Working capacity and cardiovascular response to exercise were studied in men subjected to 7-days of dry immersion. Heart rate, ventilation, and oxygen uptake and consumption were measured before immersion and on the 4th and 7th days of immersion. No significant changes in heart rate were found, but some changes in ventilation and oxygen uptake dynamics indicate that gas exchange may be altered during immersion. Physiological adaptation and individual differences are discussed.^ieng

Simple technique to evaluate on the ground the energetic expenditure of physical exercise carried out in weightlessness.

A simple method is described to evaluate the energetic expenditure of graded muscular exercises carried out on the ground in such a way as to approximate the state of space microgravity. Consumption of O2 measured at different rates of execution is shown as well as a tentative computation of the mechanical work on the basis of the accelerations impressed alternately on the mass of the trunk and upper limbs. The practicability of the method proposed under space conditions is pointed out; it does not require any ergometer, in fact, but only the fixing of the subject's feet to the floor of the vehicle.