The effect of weightlessness on the human skeletal system is one of the greatest concerns in safely extending space missions [1–11]. The ability to understand and counteract weightlessness-induced bone mineral loss will be vital to crew health and safety during and after extended-duration space sta- tion and exploration missions [1–7]. Research on bone mineral loss during space flight has gone on for more than half a century, and recent studies have shown significant progress in developing coun- termeasures that have proved to be effective, including good nutrition and exercise. We review the history of this research here and provide a summary of recent and ongoing studies, including efforts to counteract bone and calcium loss resulting from weightlessness. Unfortunately, the most obvious nutritional countermeasure—providing excess calcium—does not protect against bone loss . This result is likely related to the decreased calcium absorption observed in space flight and in ground-based models [13–16]. Phosphate supplementation was also ineffective at reducing calcium excretion . Combination therapy with calcium and phosphorus was also unsuccessful at mitigating bone loss and hypercalciuria . Other nutrients, specifically sodium, protein, potassium, vitamin K, and omega-3 fatty acids, have also been proposed and/or tested as bone loss countermeasures , and are discussed in more detail below.
Bone loss and renal stone risk are longstanding concerns for astronauts. Bone resorption brought on by spaceflight elevates urinary calcium and the risk of renal stone formation. Loss of bone calcium leads to concerns about fracture risk and increased long-term risk of osteoporosis. Bone metabolism involves many factors and is interconnected with muscle metabolism and diet. We report here bone biochemistry and renal stone risk data from astronauts on 4- to 6-month International Space Station missions. All had access to a type of resistive exercise countermeasure hardware, either the Advanced Resistance Exercise Device (ARED) or the Interim Resistance Exercise Device (iRED). A subset of the ARED group also tested the bisphosphonate alendronate as a potential anti-resorptive countermeasure (Bis+ARED). While some of the basic bone marker data have been published, we provide here a more comprehensive evaluation of bone biochemistry with a larger group of astronauts. Regardless of exercise, the risk of renal stone formation increased during spaceflight. A key factor in this increase was urine volume, which was lower during flight in all groups at all time points. Thus, the easiest way to mitigate renal stone risk is to increase fluid consumption. ARED use increased bone formation without changing bone resorption, and mitigated a drop in parathyroid hormone in iRED astronauts. Sclerostin, an osteocyte-derived negative regulator of bone formation, increased 10-15% in both groups of astronauts who used the ARED (p<0.06). IGF-1, which regulates bone growth and formation, increased during flight in all 3 groups (p<0.001). Our results are consistent with the growing body of literature showing that the hyper-resorptive state of bone that is brought on by spaceflight can be countered pharmacologically or mitigated through an exercise-induced increase in bone formation, with nutritional support. Key questions remain about the effect of exercise-induced alterations in bone metabolism on bone strength and fracture risk.
Magnesium is an essential nutrient for muscle, cardiovascular, and bone health on Earth, and during space flight. We sought to evaluate magnesium status in 43 astronauts (34 male, 9 female; 47 +/- 5 years old, mean +/- SD) before, during, and after 4-6-month space missions. We also studied individuals participating in a ground analog of space flight (head-down-tilt bed rest; n = 27 (17 male, 10 female), 35 +/- 7 years old). We evaluated serum concentration and 24-h urinary excretion of magnesium, along with estimates of tissue magnesium status from sublingual cells. Serum magnesium increased late in flight, while urinary magnesium excretion was higher over the course of 180-day space missions. Urinary magnesium increased during flight but decreased significantly at landing. Neither serum nor urinary magnesium changed during bed rest. For flight and bed rest, significant correlations existed between the area under the curve of serum and urinary magnesium and the change in total body bone mineral content. Tissue magnesium concentration was unchanged after flight and bed rest. Increased excretion of magnesium is likely partially from bone and partially from diet, but importantly, it does not come at the expense of muscle tissue stores. While further study is needed to better understand the implications of these findings for longer space exploration missions, magnesium homeostasis and tissue status seem well maintained during 4-6-month space missions.
Synchrony of the mammalian circadian clock is achieved by complex transcriptional and translational feedback loops centered on the BMAL1:CLOCK heterodimer. Modulation of circadian feedback loops is essential for maintaining rhythmicity, yet the role of transcriptional coactivators in driving BMAL1:CLOCK transcriptional networks is largely unexplored. Here, we show diurnal hepatic steroid receptor coactivator 2 (SRC-2) recruitment to the genome that extensively overlaps with the BMAL1 cistrome during the light phase, targeting genes that enrich for circadian and metabolic processes. Notably, SRC-2 ablation impairs wheel-running behavior, alters circadian gene expression in several peripheral tissues, alters the rhythmicity of the hepatic metabolome, and deregulates the synchronization of cell-autonomous metabolites. We identify SRC-2 as a potent coregulator of BMAL1:CLOCK and find that SRC-2 targets itself with BMAL1:CLOCK in a feedforward loop. Collectively, our data suggest that SRC-2 is a transcriptional coactivator of the BMAL1:CLOCK oscillators and establish SRC-2 as a critical positive regulator of the mammalian circadian clock.
Selective weighting of cutaneous receptor feedback and associated balance impairments following short duration space flight
The present study investigated the perception of low frequency (3 Hz) vibration on the foot sole and its relationship to standing balance following short duration space flight in nine astronauts. Both 3 Hz vibration perception threshold (VPT) and standing balance measures increased on landing day compared to pre-flight. Contrary to our hypothesis, a positive linear relationship between these measures was not observed; however astronauts with the most sensitive skin (lowest 3 Hz VPT) were found to have the largest sway on landing day. While the change in foot sole sensitivity does not appear to directly relate to standing balance control, an exploratory strategy may be employed by astronauts whose threshold to pressure information is lower. Understanding sensory adaptations and balance control has implications to improve balance control strategies following space flight and in sensory impaired populations on earth.
Comprehensive analysis of the skin fungal microbiota of astronauts during a half-year stay at the International Space Station
The International Space Station (ISS) is a huge manned construct located approximately 400 km above the earth and is inhabited by astronauts performing space experiments. Because the station is within a closed microgravity environment, the astronauts are subject to consistent stress. This study analyzed the temporal changes in the skin fungal microbiota of 10 astronauts using pyrosequencing and quantitative PCR assay before, during, and after their stay in the ISS. Lipophilic skin fungi, Malassezia predominated most samples regardless of the collection period, body site (cheek or chest), or subject. During their stay in the ISS, the level of Malassezia colonization changed by 7.6- +/- 7.5-fold (mean +/- standard deviation) and 9.5- +/- 24.2-fold in cheek and chest samples, respectively. At the species level, M. restricta, M. globosa, and M. sympodialis were more abundant. In the chest samples, the ratio of M. restricta to all Malassezia species increased, whereas it did not change considerably in cheek samples. Fungal diversity was reduced, and the ratio of Malassezia to all fungal colonization increased during the astronauts’ stay at the ISS. The ascomycetous yeast Cyberlindnera jadinii was detected in abundance in the in-flight sample of 5 of the 10 astronauts. The microorganism may have incidentally adhered to the skin during the preflight period and persisted on the skin thereafter. This observation suggests the ability of a specific or uncommon microorganism to proliferate in a closed environment. Our study is the first to reveal temporal changes in the skin fungal microbiota of ISS astronauts. These findings will provide information useful for maintaining the health of astronauts staying in the space environment for long periods and for preventing infection due to the human skin microbiota.
First Direct Observation of Impurity Effects on the Growth Rate of Tetragonal Lysozyme Crystals under Microgravity as Measured by Interferometry
The normal growth rates R and apparent step velocities (lateral growth rates of a spiral hillock) V of tetragonal hen egg-white lysozyme (HEWL) crystals were for the first time measured by Michelson interferometry in the international space station (as part of the NanoStep project) using commercialized HEWL samples containing 1.5% impurities. A significant increase in V under microgravity was confirmed compared to step velocities Vstep on the ground, while a decrease in R was also confirmed compared to that in the purified solution under microgravity as expected. Because of exact measurement of growth rates, kinetic analyses of R were conducted as a function of supersaturation, σ (σ ≡ ln(C/Ce), where C is the concentration; Ce is the solubility), using a spiral growth model and a two-dimensional (2D) nucleation growth model. For both models over a wide range of σ, R in the impure solution was significantly lower than that in the purified solution. The degree of the suppression of impurity effects was also evaluated using the difference in Vp and Vi, where Vp is the apparent step velocity in the purified solution, and Vi is that in the impure solution. The difference between Vp and Vi was smaller than the difference in step velocities on the ground, Vstep,p and Vstep,i, where Vstep,p is the step velocity in the purified solution, and Vstep,i is the step velocity in the impure solution.
Expression of p53-Regulated Proteins in Human Cultured Lymphoblastoid TSCE5 and WTK1 Cell Lines during Spaceflight
The aim of this study was to determine the biological effects of space radiations, microgravity, and the interaction of them on the expression of p53-regulated proteins. Space experiments were performed with two human cultured lymphoblastoid cell lines: one line (TSCE5) bears a wild-type p53 gene status, and another line (WTK1) bears a mutated p53 gene status. Under 1 gravity or microgravity conditions, the cells were grown in the cell biology experimental facility (CBEF) of the International Space Station for 8 days without experiencing the stress during launching and landing because the cells were frozen during these periods. Ground control samples were simultaneously cultured for 8 days in the CBEF on the ground for 8 days. After spaceflight, protein expression was analyzed using a PanoramaTM Ab MicroArray protein chips. It was found that p53-dependent up-regulated proteins in response to space radiations and space environment were MeCP2 (methyl CpG binding protein 2), and Notch1 (Notch homolog 1), respectively. On the other hand, p53-dependent down-regulated proteins were TGF-β, TWEAKR (tumor necrosis fac- tor-like weak inducer of apoptosis receptor), phosho-Pyk2 (Proline-rich tyrosine kinase 2), and 14-3-3θ/τ which were affected by microgravity, and DR4 (death receptor 4), PRMT1 (protein arginine methyltrans- ferase 1) and ROCK-2 (Rho-associated, coiled-coil containing protein kinase 2) in response to space radi- ations. ROCK-2 was also suppressed in response to the space environment. The data provides the p53- dependent regulated proteins by exposure to space radiations and/or microgravity during spaceflight. Our expression data revealed proteins that might help to advance the basic space radiation biology.
Manned space flight induces a reduction in immune competence among crew and is likely to cause deleterious changes to the composition of the gastrointestinal, nasal, and respiratory bacterial flora, leading to an increased risk of infection. The space flight environment may also affect the susceptibility of microorganisms within the spacecraft to antibiotics, key components of flown medical kits, and may modify the virulence characteristics of bacteria and other microorganisms that contaminate the fabric of the International Space Station and other flight platforms. This review will consider the impact of true and simulated microgravity and other characteristics of the space flight environment on bacterial cell behavior in relation to the potential for serious infections that may appear during missions to astronomical objects beyond low Earth orbit.
Three-dimensional structure of phosphoribosyl pyrophosphate synthetase from E. coli at 2.71 Å resolution
Phosphoribosyl pyrophosphate synthetase from Escherichia coli was cloned, purified, and crystal- lized. Single crystals of the enzyme were grown under microgravity. The X-ray diffraction data set was col- lected at the Spring-8 synchrotron facility and used to determine the three-dimensional structure of the enzyme by the molecular-replacement method at 2.71 Å resolution. The active and regulatory sites in the molecule of E. coli phosphoribosyl pyrophosphate synthetase were revealed by comparison with the homol- ogous protein from Bacillus subtilis, the structure of which was determined in a complex with functional ligands. The conformations of polypeptide-chain fragments surrounding and composing the active and reg- ulatory sites were shown to be identical in both proteins.