Long-Term Persistence of Increased Number of γH2AX+ Peripheral Blood Lymphocytes in Monkeys Exposed to Negative Factors of Space Flights: Ionizing Radiation and Simulated Hypogravity.

We studied the influence of ionizing radiation and hypogravity as negative factors of space flights on DNA damage in peripheral blood lymphocytes of rhesus monkeys at different times after exposure (from 1 to 446 days). The proportion of cells with high numbers of DNA double-strand breaks (DSB), positive for the surrogate DSB marker-protein γH2AX, was monitored using flow cytometry. Some animals were exposed to 7-day antiorthostatic hypokinesia simulating hypogravity, the others to a combined effect of antiorthostatic hypokinesia, whole-body γ-irradiation (2.34 cGy/h, dose 1 Gy), and irradiation of the head with 12C ions (450 MeV, dose 1 Gy). Exposure to antiorthostatic hypokinesia led to a significant increase in the proportion of γH2AX+ lymphocytes only on the first day after exposure, whereas after combined exposure, increased numbers of damaged lymphocytes were recorded up to 42 days after exposure.

Stumbling reactions in hypo and hyper gravity - muscle synergies are robust across different perturbations of human stance during parabolic flights.

The control of bipedal stance and the capacity to regain postural equilibrium after its deterioration in variable gravities are crucial prerequisites for manned space missions. With an emphasize on natural orthograde posture, computational techniques synthesize muscle activation patterns of high complexity to a simple synergy organization. We used nonnegative matrix factorization to identify muscle synergies during postural recovery responses in human and to examine the functional significance of such synergies for hyper-gravity (1.75 g) and hypo-gravity (0.25 g). Electromyographic data were recorded from leg, trunk and arm muscles of five human exposed to five modes of anterior and posterior support surface translations during parabolic flights including transitional g-levels of 0.25, 1 and 1.75 g. Results showed that in 1 g four synergies accounted for 99% of the automatic postural response across all muscles and perturbation directions. Each synergy in 1 g was correlated to the corresponding one in 0.25 and 1.75 g. This study therefore emphasizes the similarity of the synergy organization of postural recovery responses in Earth, hypo- and hyper-gravity conditions, indicating that the muscle synergies and segmental strategies acquired under terrestrial habits are robust and persistent across variable and acute changes in gravity levels.

An investigation of shock wave therapy and low-intensity pulsed ultrasound on fracture healing under reduced loading conditions in an ovine model.

The use of shock wave therapy (SWT) and low-intensity pulsed ultrasound (LIPUS) as countermeasures to the inhibited fracture healing experienced during mechanical unloading was investigated by administering treatment to the fracture sites of mature, female, Rambouillet Columbian ewes exposed to partial mechanical unloading or full gravitational loading. The amount of fracture healing experienced by the treatment groups was compared to controls in which identical surgical and testing protocols were administered except for SWT or LIPUS treatment. All groups were euthanized after a 28-day healing period. In vivo mechanical measurements demonstrated no significant alteration in fixation plate strains between treatments within either partial unloading group. Similarly, DXA BMD and 4-point bending stiffness were not significantly altered following either treatment. µCT analyses demonstrated lower callus bone volume for treated animals (SWT and LIPUS, p < 0.01) in the full gravity group but not between reduced loading groups. Callus osteoblast numbers as well as mineralized surface and bone formation rate were significantly elevated to the level of the full gravity groups in the reduced loading groups following both SWT and LIPUS. Although no increase in 4-week mechanical strength was observed, it is possible that an increase in the overall rate of fracture healing (i.e., callus strength) may be experienced at longer time points under partial loading conditions given the increase in osteoblast numbers and bone formation parameters following SWT and LIPUS. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:921-929, 2018.

Effect of Long-Term Simulated Microgravity on Immune System and Lung Tissues in Rhesus Macaque.

We evaluated different lymphocyte populations and levels of plasma cytokines in peripheral blood as well as inflammatory infiltration and expressions of cytokines in lung tissues derived from macaque under long-term stimulated microgravity through being suspended in an antiorthostatic position so as to identify relevant immune parameters and to understand potential mechanisms of lung injury. Fifteen healthy male rhesus macaques were randomly divided into groups 1 (control, n = 5), groups 2 (head-down tilting for 6 weeks, n = 5), and groups 3 (head-down tilting for 6 weeks and recovery for 4 weeks, n = 5). Lymphocyte subsets in peripheral blood were analyzed using flow cytometry and the concentrations of 14 cytokines in plasma were measured with Luminex multiplexing technology. HE staining and transmission electron microscopy were employed to investigate the morphologies and subcellular structures of lung tissues. Immunohistochemistry and real-time PCR were employed to explore mRNA and protein expressions of cytokines in lung tissues. Immunohistochemical demonstrations were detected for CD3, CD4, CD8 T lymphocytes, CD20 B lymphocytes, and CD68 macrophages in lung tissues. Compared to group 1, groups 2 and 3 showed a decrease in the percentage of CD2+T cells, CD2+CD4+T helper cells, and CD2+CD8+cytotoxic T cells as well as an increase in the expression of CD95 on the surface of T lymphocytes in peripheral blood. The serum cytokine levels of IL-18 and TNF-α were increased in group 2 when compared to groups 1 and 3. HE and TEM observed changes in the structure and ultrastructure of lung tissues in groups 2 and 3. The number of CD3+T cell, CD4+T cell, CD8+T cells, and CD68+macrophage and the expression levels of IL-1ß, IL-6, and IL-18 in lung tissues were increased in groups 2 when compared with groups 1 and 3. Our data suggested that long-term microgravity might alter the functions of immune system and cause lung damage, changing lymphocyte distribution and functions as well as cytokine production.

The relationship between widespread changes in gravity and cerebral blood flow.

OBJECTIVES: We investigated the dose-effect relationship between wide changes in gravity from 0 to 2.0 Gz (Δ0.5 Gz) and cerebral blood flow (CBF), to test our hypothesis that CBF has a linear relationship with levels of gravity. SUBJECTS AND METHODS: Ten healthy seated men were exposed to 0, 0.5, 1.0, 1.5, and 2.0 Gz for 21 min, by using a tilt chair and a short-arm human centrifuge. Steady-state CBF velocity (CBFV) in the middle cerebral artery by transcranial Doppler ultrasonography, mean arterial pressure (MAP) at the heart level (MAPHeart), heart rate, stroke volume, cardiac output and respiratory conditions were obtained for the last 6 min at each gravity level. Then, MAP in the middle cerebral artery (MAPMCA), reflecting cerebral perfusion pressure, was estimated. RESULTS: Steady-state CBFV decreased stepwise from 0.5 to 2.0 Gz. Steady-state heart rate, stroke volume, estimated MAPMCA and end-tidal carbon dioxide pressure (ETCO2) also changed stepwise from hypogravity to hypergravity. On the other hand, steady-state MAPHeart and cardiac output did not change significantly. Steady-state CBFV positively and linearly correlated with estimated MAPMCA and ETCO2 in most subjects. CONCLUSION: The present study demonstrated stepwise gravity-induced changes in steady-state CBFV from 0.5 to 2.0 Gz despite unchanged steady-state MAPHeart. The combined effects of reduced MAPMCA and ETCO2 likely led to stepwise decreases in CBFV. We caution that a mild increase in gravity from 0 to 2.0 Gz reduces CBF, even if arterial blood pressure at the heart level is maintained.

Swimming and bone: Is low bone mass due to hypogravity alone or does other physical activity influence it?

UNLABELLED: Swimming during adolescence has shown neutral or even negative effects on bone mass. Nevertheless, it is still unknown if these effects are due to swimming or to other factors, such as sedentary behaviors. INTRODUCTION: Three objectives were described (1) to measure objective physical activity (PA) additional to swimming performed by adolescent swimmers (SWI) and compare it to that performed by normo-active controls (CG), (2) to describe the relationship between objectively measured PA and bone mass, and (3) to compare bone mass of swimmers that meet the World Health Organization PA guidelines (active) WHO and those that do not (inactive). METHODS: A total of 71 SWI (33 females) and 41 CG (17 females) wore an accelerometer for at least 4 days. PA was expressed as the amount of time (minutes/day) in each intensity [sedentary/light/moderate or vigorous (VPA), and the sum of moderate and vigorous (MVPA)]. Using the cutoff points proposed by Vanhelst et al. SWI were classified as active or inactive according to whether they reached 60 min of weight-bearing MVPA per day or not. Bone mineral density (BMD) was measured by dual energy X-ray absorptiometry, and bone strength values were calculated with peripheral quantitative computed tomography. Differences in PA intensities were calculated between SWI and CG. The relation of VPA to bone mass was studied in the SWI. RESULTS: Male-SWI spend less time in VPA and MVPA than male-GC, which partly explains the lower BMD values in SWI than CG. CONCLUSION: Swimming may displace weight-bearing VPA with serious implications on bone health.

Computational characterization of fracture healing under reduced gravity loading conditions.

The literature is deficient with regard to how the localized mechanical environment of skeletal tissue is altered during reduced gravitational loading and how these alterations affect fracture healing. Thus, a finite element model of the ovine hindlimb was created to characterize the local mechanical environment responsible for the inhibited fracture healing observed under experimental simulated hypogravity conditions. Following convergence and verification studies, hydrostatic pressure and strain within a diaphyseal fracture of the metatarsus were evaluated for models under both 1 and 0.25 g loading environments and compared to results of a related in vivo study. Results of the study suggest that reductions in hydrostatic pressure and strain of the healing fracture for animals exposed to reduced gravitational loading conditions contributed to an inhibited healing process, with animals exposed to the simulated hypogravity environment subsequently initiating an intramembranous bone formation process rather than the typical endochondral ossification healing process experienced by animals healing in a 1 g gravitational environment. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1206-1215, 2016.

INTRAOCULAR PRESSURE AND EYE HYDRODYNAMICS DURING BRIEF HEAD-DOWN TILT.

Intraocular pressure (IOP) and eye hydrodynamics (aqueous outflow easiness rate (C) and moisture chamber production (F)) were studied in 9 adult volunteers subjected to the hypogravity effects of head-down tilt (HDT) at -15° to the horizontal plane. The volunteers stayed in the horizontal and tilted positions for 10 minutes. IOP was measured according to Maklakov (tonometer 5 g), C and F - according to Nesterov (simplified tonography). In parallel, heart rate (HR) and systolic and diastolic blood pressures (SBP and DBP) were measured in the sitting, lying and tilted positions. In HDT IOP rose 10.3 % (p < 0.05) and C reduced 60 % (p < 0.05); F showed an uncertain trend down by 59 % (p > 0.05). Increase of the Bekker coefficient by 168 % (p < 0.05) could testify interconnection of the increased IOP and impaired moisture outflow. Moreover, in HDT DBP showed a rise while HR decreased. These results suggest that during brief tilt- down IOP increases not only because of a greater filling of the choroid vessels, but also because of retarded outflow of the intraocular fluid. The downward trend in fluid production can be a compensatory reaction to increased'IOP.

Lumbar puncture during spaceflight: operational considerations, constraints, concerns, and limitations.

Lumbar puncture (LP) is a commonly performed low-risk procedure terrestrially, used diagnostically for evaluation of cerebrospinal fluid (CSF) pressure as well as for collection of CSF for analysis. NASA is investigating noninvasive means for measurement of intracranial pressure (ICP) to assess the potential contribution of elevated intracranial pressures to recently reported changes in astronauts' visual acuity and eye anatomy, known collectively as the Visual Impairment/Intracranial Pressure risk. However, many of these noninvasive technologies are still under development, have limited clinical validation, are several years away from being ready for in-flight use, or only provide qualitative rather than quantitative ICP values. Therefore, performance of in-flight LPs, as part of crewmember evaluation, has also been considered by NASA. This manuscript summarizes the unique operational considerations, constraints, concerns, and limitations of using traditional LP as an adjunct or as an alternative to noninvasive ICP measurements during spaceflight.

Flat spin and negative Gz in high-altitude free fall: pathophysiology, prevention, and treatment.

INTRODUCTION: Red Bull Stratos was a commercial program that brought a test parachutist protected by a full pressure suit to 127,852 ft (38,964 m), via a stratospheric balloon with a pressurized capsule, from which he free fell and subsequently parachuted to the ground. In light of the uniqueness of the operation and the medical threats faced, medical protocols specific to distinctive injury patterns were developed. One unique threat was that of a flat spin during free fall with resultant exposure to -Gz (toe-to-head) acceleration. In preparation for stratospheric free fall, the medical team conducted a review of the literature on the spectrum of human and animal injury patterns attributable to -Gz exposures. Based on the findings, an emergency medical field response protocol was developed for the rapid assessment, diagnosis, and treatment of individuals suspected of -Gz injury. METHODS: A systematic review was conducted on available literature on human and animal studies involving significant -Gz exposure, with subsequent development of an applicable field treatment protocol. RESULTS: The literature review identified pathophysiologic processes and mitigation strategies that were used to develop a prevention and treatment protocol, outlining appropriate interventions using current best medical practices. A medical field treatment protocol was successfully established for the high-altitude balloon program. DISCUSSION: Available literature provided insight into best medical practices for the prevention and treatment of significant -Gz exposures during high-altitude parachute activity. Using the protocol developed for the field medical response, injuries from sustained -Gz exposure can be effectively managed in similar high-altitude and space operations.

[Myelination disorders in mechanism of hypogravity motor syndrome development].

When modeling effects of hypogravitation by the method of hindlimb unloading in rats the area of cross-section in lumbar part of a spinal cord was found to reduce. The analysis of spinal cord slides showed that these changes are associated with a decrease in the area of white substance of a spinal cord. Data obtained are consistent with our previous observation of a decrease in expression of the genes encoding myelin proteins. Results of our researches give the good reasons to believe that miyelinization failure in CNS is one of the factors that underlie the development of hypogravitational motor syndrome.

Human mesenchymal stem cells are sensitive to abnormal gravity and exhibit classic apoptotic features.

The aim of the present study was to investigate the effects of abnormal gravity on human mesenchymal stem cells (hMSCs). Strong magnetic field and magnetic field gradient generate a magnetic force that can add to or subtract from the gravitational force. In this study, this is defined as a high-magneto-gravitational environment (HMGE). The HMGE provides three apparent gravity levels, i.e. hypogravity (µg), hypergravity (2g) and normal gravity with strong magnetic field (1g) conditions. After hMSCs were subject to HMGE for 12 h, the proliferation, morphology, structure and apoptosis were investigated. Results showed that the proliferation of hMSCs was inhibited under µg condition. The abnormal gravity induced morphologic characteristics of apoptosis cells, such as cell shrinkage, membrane blebbing, nuclear chromatin condensation and margination, decreased cell viability, and increased caspase-3/7 activity. The rate of apoptosis under µg condition is up to 56.95%. The F-actin stress fibers and microtubules were disrupted under abnormal gravity condition. Under µg-condition, the expression of p53 at mRNA and protein levels was up-regulated more than 9- and 6 folds, respectively. The Pifithrin-α, an specific inhibitor of p53, inhibited the apoptosis and prevented the disruption of cytoskeleton induced by abnormal gravity. These results implied that hMSCs were sensitive to abnormal gravity and exhibited classic apoptotic features, which might be associated with p53 signaling.

[Role of calcium-dependent mechanisms in the development of atrophy in postural muscle deprived of gravitational loading].

Downsizing of muscle fibers, decline of their contractility and alteration of the myosin phenotype towards fast isoforms prevalence are ranked among the main consequences of gravitational unloading of postural muscles. Role of Ca2+ ions in these processes is the subject of the article. Authors revealed increase in the Ca2+ content in myofibers of resting m. soleus of mice following hindlimb suspension. It has been also reported earlier that nifedipine, a specific blocker of calcium L-channels, can prevent this increase acting on the central and local controls. Therefore, a supposition can be made that activation of the dehydropyridine Ca channels is responsible for Ca2+ rest accumulation during gravitational unloading. The calcium-dependent signaling pathways may be thought of as key players in a number of developments due to gravitational unloading. To begin with, already in early 1990s this role in the hypogravity-induced muscle atrophy was ascribed to calpains, Ca-dependent proteinases which was clearly demonstrated later. We observed maintenance of the relative titin and nebulin contents in unloaded m. soleus associated with the effect of Ca chelators. Also, nifedipine injection reduced significantly the growth of MHC fast isoforms expression during gravitational unloading equally on the RNA and protein levels. To bring to light the place of calcineurin/NFAT signaling system in the MHC phenotype alteration, we undertook inhibition of this pathway by injection of cyclosporine A. This resulted in getting evidence of the stabilizing function of this pathway as it opposes to the myosin phenotype transformation to the fast one.

[Physiological changes in microgravity]. / Modifications physiologiques en microgravité.

Microgravity affects the different organ systems to various degrees. In particular, a thoracocephalic fluid shift occurs through suppression of the hydrostatic gradient. Sensory conflict leads to space motion sickness, which is frequent early in the flight, and the musculo-skeletal system is perturbed by the lack of stimuli. The respiratory system seems to be less affected. These changes do not seem to impair health or performance. Humans seem able to adapt to long-term exposure to microgravity, but disorders can occur on return to Earth. In-flight preventive measures need be developed.

Gravitational unloading induces an anti-angiogenic phenotype in human microvascular endothelial cells.

Creating conditions similar to those occurring during exposure of cells to microgravity modulates endothelial functions. We have previously demonstrated that human macrovascular endothelial cells in simulated hypogravity proliferate faster than controls, partly because they downregulate interleukin 1alpha. On the contrary, murine microvascular endothelial cells are growth inhibited in simulated hypogravity, and this is due, at least in part, to the decrease of interleukin 6. Since endothelial cells are very heterogeneous and differences between various species have been reported, we exposed human microvascular cells to gravitational unloading and found that it retards cell growth without affecting cell migration. Interestingly, we detected the induction of Tissue Inhibitor of Metalloprotease-2, which inhibits endothelial growth in vitro and angiogenesis in vivo. Together with the finding that hypogravity stimulates the synthesis of nitric oxide, involved also in neovascularization, our results underscore a modulation of the angiogenic properties of microvascular human endothelial cells. We also show that hypogravity inhibits proteasome activity, thus suggesting that post-translational mechanisms are involved in the adaptations of these cells to hypogravity. These results underscore that hypogravity differently impacts on micro- and macro-vascular human endothelial cells. In particular, these results may shed some light on the molecular mechanisms contributing to the impairment of angiogenesis observed in different models in space. Our data might also explain why bioengineered tissues to be used for regenerative medicine fail to neovascularize when assembled in simulated hypogravity.

Bone height measurements around a dental implant after a 6-month space flight: a case report.

PURPOSE: In space, astronauts are subject to microgravity, which reduces skeletal loading and osteoblast function and can cause bone resorption and a decrease in bone density. No known research to date has studied the effect of microgravity on dental implants. This study evaluated peri-implant bone changes around a dental implant placed in a French astronaut who spent 6 months in Russia's Mir Space Station. MATERIALS AND METHODS: Measurements were performed by 2 examiners before the flight (baseline), after the flight (stage 1), and following a recovery period (stage 2). Standardized periapical radiographs were taken, and data were recorded using a photomicroscope and a measuring scale. RESULTS: Cumulatively, the implant sustained 0.43 mm of mesial bone gain and 0.31 mm of distal bone loss. DISCUSSION: The observed peri-implant bone height changes were within normal limits and the implant appeared very stable during the course of this study. CONCLUSION: Peri-implant bone levels remained stable after 6 months in microgravity, and the implant continued to function without complications.

Seven-degree head-down tilt reduces choroidal pulsatile ocular blood flow.

INTRODUCTION: We investigated the effects of head-down tilt (HDT), which simulates microgravity during spaceflights, on the choroidal pulsatile ocular blood flow (POBF). This investigation is important because alterations in the choroidal blood flow can affect the function of retinal rods and cones that rely totally on the choroid for metabolites. METHODS: Nineteen healthy adults between 20 and 38 yr of age participated in this study. The POBF was compared for: 1) baseline, wherein subjects were declined 30 degrees from vertical; 2) microgravity simulation where subjects were in a 7 degrees HDT for 2 min; 3) 90 min of the 7 degrees HDT; and 4) recovery, i.e., back at 30 degrees for 2 min. RESULTS: The group averaged POBF (Mean +/- SEM values: 828.43 +/- 48.88 microL x min(-1)) decreased immediately during the 2-min microgravity simulation (582.18 +/- 43.62 microL x min(-1)), remained at that inferior level at the 90-min mark of HDT (542.26 +/- 45.35 microL x min(-1)), and came back toward baseline POBF during the recovery period (760.11 +/- 46.03 microL x min(-1)) (p = 0.0001). DISCUSSION: The results show that simulated-microgravity of relatively short duration induces retinal hypoperfusion throughout the microgravity interval through the reduction in the POBF. This finding may have important implications regarding visual performance in space crewmembers placed in prolonged microgravity environments.

The Bellagio Report: Cardiovascular risks of spaceflight: implications for the future of space travel.

BACKGROUND: Long-duration space missions, as well as emerging civilian tourist space travel activities, prompted review and assessment of data available to date focusing on cardiovascular risk and available risk mitigation strategies. The goal was the creation of tools for risk priority assessments taking into account the probability of the occurrence of an adverse cardiovascular event and available and published literature from spaceflight data as well as available risk mitigation strategies. METHODS: An international group of scientists convened in Bellagio, Italy, in 2004 under the auspices of the Aerospace Medical Association to review available literature for cardiac risks identified in the Bioastronautics Critical Path Roadmap (versions 2000, 2004). This effort led to the creation of a priority assessment framework to allow for an objective assessment of the hazard, probability of its occurrence, mission impact, and available risk mitigation measures. RESULTS/CONCLUSIONS: Spaceflight data are presented regarding evidence/ no evidence of cardiac dysrhythmias, cardiovascular disease, and cardiac function as well as orthostatic intolerance, exercise capacity, and peripheral resistance in presyncopal astronauts compared to non-presyncopal astronauts. Assessment of the priority of different countermeasures was achieved with a tabular framework with focus on probability of occurrence, mission impact, compliance, practicality, and effectiveness of countermeasures. Special operational settings and circumstances related to sensitive portions of any mission and the impact of environmental influences on mission effectiveness are addressed. The need for development of diagnostic tools, techniques, and countermeasure devices, food preparation, preservation technologies and medication, as well as an infrastructure to support these operations are stressed. Selected countermeasure options, including artificial gravity and pharmacological countermeasures need to be systematically evaluated and validated in flight, especially after long-duration exposures. Data need to be collected regarding the emerging field of suborbital and orbital civilian space travel, to allow for sound risk assessment.