Evaluation of upper body muscle activity during cardiopulmonary resuscitation performance in simulated microgravity

Performance of efficient single-person cardiopulmonary resuscitation (CPR) is vital to maintain cardiac and cerebral perfusion during the 2–4 min it takes for deployment of advanced life support during a space mission. The aim of the present study was to investigate potential differences in upper body muscle activity during CPR performance at terrestrial gravity (+1Gz) and in simulated microgravity (μG). Muscle activity of the triceps brachii, erector spinae, rectus abdominis and pectoralis major was measured via superficial electromyography in 20 healthy male volunteers. Four sets of 30 external chest compressions (ECCs) were performed on a mannequin. Microgravity was simulated using a body suspension device and harness; the Evetts–Russomano (ER) method was adopted for CPR performance in simulated microgravity. Heart rate and perceived exertion via Borg scores were also measured. While a significantly lower depth of ECCs was observed in simulated microgravity, compared with +1Gz, it was still within the target range of 40–50 mm. There was a 7.7% decrease of the mean (±SEM) ECC depth from 48 ± 0.3 mm at +1Gz, to 44.3 ± 0.5 mm during microgravity simulation (p < 0.001). No significant difference in number or rate of compressions was found between the two conditions. Heart rate displayed a significantly larger increase during CPR in simulated microgravity than at +1Gz, the former presenting a mean (±SEM) of 23.6 ± 2.91 bpm and the latter, 76.6 ± 3.8 bpm (p < 0.001). Borg scores were 70% higher post-microgravity compressions (17 ± 1) than post +1Gz compressions (10 ± 1) (p < 0.001). Intermuscular comparisons showed the triceps brachii to have significantly lower muscle activity than each of the other three tested muscles, in both +1Gz and microgravity. As shown by greater Borg scores and heart rate increases, CPR performance in simulated microgravity is more fatiguing than at +1Gz. Nevertheless, no significant difference in muscle activity between conditions was found, a result that is favourable for astronauts, given the inevitable muscular and cardiovascular deconditioning that occurs during space travel.     

Signs of Müller cell gliotic response found in the retina of newts exposed to real and simulated microgravity

The effects of real and simulated microgravity on the eye tissue regeneration of newts were investigated. For the first time changes in Müller glial cells in the retina of eyes regenerating after retinal detachment were detected in newts exposed to clinorotation. The cells divided, were hypertrophied, and their processes were thickened. Such changes suggested reactive gliosis and were more significant in animals exposed to rotation when compared with desk-top controls. Later experiments onboard the Russian biosatellite Bion-11 showed similar changes in the retinas that were regenerating in a two-week spaceflight. In the Bion-11 animals, GFAP, the major structural protein of retinal macroglial cells, was found to be upregulated. In a more recent experiment onboard Foton-M3 (2007), GFAP expression in retinas of space-flown, ground control (kept at 1 g), and basal control (sacrificed on launch day) newts was quantified, using microscopy, immunohistochemistry, and digital image analysis. A low level of immunoreactivity was observed in basal controls. In contrast, retinas of space-flown animals showed greater GFAP immunoreactivity associated with both an increased cell number and a higher thickness of intermediate filaments. This, in turn, was accompanied by up-regulation of stress protein (HSP90) and growth factor (FGF2) expressions. It can be postulated that such a response of Müller cells was to mitigate the retinal stress in newts exposed to microgravity. Taken together, the data suggest that the retinal population of macroglial cells could be sensitive to gravity changes and that in space it can react by enhancing its neuroprotective function.     

Simulated microgravity inhibits cell wall regeneration of Penicillium decumbens protoplasts

This work compares cell wall regeneration from protoplasts of the fungus Penicillium decumbens under rotary culture (simulated microgravity) and stationary cultures. Using an optimized lytic enzyme mixture, protoplasts were successfully released with a yield of 5.3 × 10⁵ cells/mL. Under simulated microgravity conditions, the protoplast regeneration efficiency was 33.8%, lower than 44.9% under stationary conditions. Laser scanning confocal microscopy gave direct evidence for reduced formation of polysaccharides under simulated conditions. Scanning electron microscopy showed the delayed process of cell wall regeneration by simulated microgravity. The delayed regeneration of P. decumbens cell wall under simulated microgravity was likely caused by the inhibition of polysaccharide synthesis. This research contributes to the understanding of how gravitational loads affect morphological and physiological processes of fungi.     

Upregulation of erythropoietin receptor in UT-7/EPO cells inhibits simulated microgravity-induced cell apoptosis

Hematopoietic progenitor cell proliferation can be altered in either spaceflight or under simulated microgravity experiments on the ground, however, the underlying mechanism remains unknown. Our previous study showed that exposure of the human erythropoietin (EPO)-dependent leukemia cell line UT-7/EPO to conditions of simulated microgravity significantly inhibited the cellular proliferation rate and induced cell apoptosis. We postulated that the downregulation of the erythropoietin receptor (EPOR) expression in UT-7/EPO cells under simulated microgravity may be a possible reason for microgravity triggered apoptosis. In this paper, a human EPOR gene was transferred into UT-7/EPO cells and the resulting expression of EPOR on the surface of UT-7/EPO cells increased approximately 61% (p < 0.05) as selected by the antibiotic G418. It was also shown through cytometry assays and morphological observations that microgravity-induced apoptosis markedly decreased in these UT-7/EPO–EPOR cells. Thus, we concluded that upregulation of EPOR in UT-7/EPO cells could inhibit the simulated microgravity-induced cell apoptosis in this EPO dependent cell line.     

Simulated microgravity inhibits the proliferation of K562 erythroleukemia cells but does not result in apoptosis

Astronauts and experimental animals in space develop the anemia of space flight, but the underlying mechanisms are still unclear. In this study, the impact of simulated microgravity on proliferation, cell death, cell cycle progress and cytoskeleton of erythroid progenitor-like K562 leukemia cells was observed. K562 cells were cultured in NASA Rotary Cell Culture System (RCCS) that was used to simulate microgravity (at 15 rpm). After culture for 24 h, 48 h, 72 h, and 96 h, the cell densities cultured in RCCS were only 55.5%, 54.3%, 67.2% and 66.4% of the flask-cultured control cells, respectively. The percentages of trypan blue-stained dead cells and the percentages of apoptotic cells demonstrated no difference between RCCS-cultured cells and flask-cultured cells at every time points (from 12 h to 96 h). Compared with flask-cultured cells, RCCS culture induced an accumulation of cell number at S phase concomitant with a decrease at G0/G1 and G2/M phases at 12 h. But 12 h later (from 24 h to 60 h), the distribution of cell cycle phases in RCCS-cultured cells became no difference compared to flask-cultured cells. Consistent with the changes of cell cycle distribution, the levels of intercellular cyclins in RCCS-cultured cells changed at 12 h, including a decrease in cyclin A, and the increasing in cyclin B, D1 and E, and then (from 24 h to 36 h) began to restore to control levels. After RCCS culture for 12–36 h, the microfilaments showed uneven and clustered distribution, and the microtubules were highly disorganized. These results indicated that RCCS-simulated microgravity could induce a transient inhibition of proliferation, but not result in apoptosis, which could involve in the development of space flight anemia. K562 cells could be a useful model to research the effects of microgravity on differentiation and proliferation of hematopoietic cells.     

The effects of simulated microgravity on the seminiferous tubules of rats

Space flight has been shown to have many adverse effects on various systems throughout the body. Because the opportunity to place research animals on board a Space Shuttle or the International Space Station is infrequent, various techniques have been designed to simulate the effects of microgravity in Earth based laboratories. A commonly used technique is known as antiorthostatic suspension, also often referred to as hind limb suspension. In this technique the hind portion of the animal is raised so that its hind limbs are non-weight bearing. This places the animal in roughly a 30° head down tilt position. This results in cephalic fluid shifts similar to those seen in actual space flight. This technique has also been shown to mimic other physiological parameters that are affected during space flight. This study examined testicular tissue from rats subjected to a 7 day antiorthostatic suspension. This tissue was acquired through a tissue sharing program and some of the experimental animals were injected with Interleukin 1 receptor antagonist (IL-1ra) which was hoped to ameliorate some of the effects of antiorthostatic suspension. The injection of IL-1ra was not expected to have any effect on testicular tissue, however this tissue was included in the morphological and statistical analysis to conduct a more complete study. All tissues were embedded in paraffin, sectioned, and stained using standard H&E staining. The tissue was then qualitatively ranked according to the “health” of the seminiferous tubules. Our findings indicate that 7 days of antiorthostatic suspension had adverse effects on the tissue that comprises the walls of the seminiferous tubules. It has long been known that antiorthostatic suspension has deleterious effects on testicular tissue, however this research indicates that these effects occur much faster than indicated by previous researchers. This is a significant finding because it indicates that meaningful earth based studies in this area can be carried out in a shorter time span. This could result in more studies per year as well as saving money by avoiding longer than necessary animal suspensions. This is especially important as we enter an era when, without Space Shuttle, flight opportunities will become scarce. These antiorthostatic suspension studies indicate that space flight, even short duration spaceflight, may have harmful effects on the seminiferous tubules and blood-testis barrier of astronauts.     

In vitro effects of simulated microgravity on Sertoli cell function

With the advent of space flights questions concerning the effects of microgravity (0×G) on human reproductive physiology have received great attention. The aim of this study was to evaluate the influence of 0×G on Sertoli cells. A Sertoli cell line from mouse testis (42GPA9) was analyzed for cytoskeletal and Sex Hormone Binding Globilin (SHBG) changes by immunohistochemistry, for antioxidant content by RT-PCR and for culture medium lactate concentrations by protein chemistry. Cells were cultured for 6, 24 and 48 h on a three-dimensional Random Positioning Machine (3D-RPM); static controls (1×G) were positioned on the supporting frame. At the end of each experiment, cultured cells were either fixed in paraformaldehyde or lysed and RNA-extracted or used for culture medium lactate measurements as needed. At 0×G, Sertoli cytoskeleton became disorganized, microtubules fragmented and SHBG undetectable already after 24 h, with alterations worsening by 48 h. It was evident that various antioxidant systems appreciably increased during the first 24 h but significantly decreased at 48 h. No changes occurred in the 1×G samples. Initially, 0×G seemed to disturb antioxidant protection strategies allowing the testes to support sperm production, thus generating an aging-like state of oxidative stress. Lactate production at 0×G slightly decreased after 24 h. Further experiments are needed in space to investigate upon steroidogenesis and germ cell differentiation within the testis, to rule out male infertility as a possible consequence, which could be a problem, as life expectancy increases.     

Effects of simulated weightlessness on the intestinal mucosal barrier of rats

This study employed a rat tail-suspension model to investigate the effects of simulated weightlessness on the intestinal mucosal barrier. Twenty-four Wistar rats were randomly divided into control (CON), 14-day tail-suspension (SUS-14d), and 21-day tail-suspension (SUS-21d) groups (n = 8 per group). Expression of occludin and zonula occludins-1 (ZO-1), proteins of the tight junction (TJ), in the intestinal mucosa was measured by immunohistochemical analysis, Western blotting, and mRNA fluorescent quantitation PCR. Plasma concentrations of diamine oxidase (DAO) and d-lactate were determined using an enzymatic spectrophotometric assay. Expression of occludin and ZO-1 was reduced in the SUS-14d and SUS-21d groups as compared to the CON group, with lowest expression observed in the SUS-21d group (P < 0.01). Examination by transmission electron microscopy (TEM) of the jejunal epithelium revealed increased intercellular space, decreased TJ and desmosome densities, and destruction of microvilli in the SUS-14d and SUS-21d groups. Plasma DAO and d-lactate concentrations in the SUS-21d group were higher than those in SUS-14d group and significantly higher than those in the CON group (P < 0.01). In all three groups, the expression of occludin and ZO-1 was found to correlate negatively with DAO (P < 0.01) and d-lactate (P < 0.01) concentrations. It is concluded that simulated weightless results in down-regulation of expression of TJ proteins in the rat intestinal mucosa. Simulated weightlessness is proposed to increase intestinal permeability through damage to the TJ.     

Involvement of nitric oxide in the mechanism of biochemical alterations induced by simulated microgravity in Microcystis aeruginosa

Simulated microgravity (SMG) can inhibit proliferation and enhance microcystin production of Microcystis aeruginosa. We investigated the role of nitric oxide (NO) in regulating the SMG induced changes of proliferation, photochemical system II photochemical activity, pigment, soluble protein and microcystin production in M. aeruginosa. M. aeruginosa was exposed to 0.1 mM sodium nitroprusside (SNP, NO donor) or 0.02 mM 2-(4-carboxyphenyl)-4, 4, 5, 5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO, NO scavenger) alone or in combination with SMG for 48 h. SMG and SNP inhibited the growth of M. aeruginosa while c-PTIO had no effect on cell number. As to yield, the negative effect of SMG was augmented by SNP and suppressed by c-PTIO. The intracellular concentrations of chlorophyll a, carotenoid, phycocyanin, soluble protein and microcystin were increased by SMG after 48 h. The effects of SMG on these metabolic processes could be enhanced by SNP and be partly eliminated by c-PTIO. Moreover, SNP and c-PTIO only functioned in these biochemical processes under SMG, unlike in the regulation of cell proliferation and yield. These results showed that the effects of SMG could be enhanced by adding exogenous NO and be mitigated by scavenging endogenous NO, revealing the involvement of NO in the changes in biochemistry processes induced by SMG in M. aeruginosa.     

Simulation analysis for effects of bone loss on acceleration tolerance of human lumbar vertebra

The purpose of the present study was to analyze and predict the changes in acceleration tolerance of human vertebra as a result of bone loss caused by long-term space flight. A human L3–L4 vertebra FEM model was constructed, in which the cancellous bone was separated, and surrounding ligaments were also taken into account. The simulation results demonstrated that bone loss has more of an effect on the acceleration tolerance in x-direction. The results serve to aid in the creation of new acceleration tolerance standards, ensuring astronauts return home safely after long-term space flight. This study shows that more attention should be focused on the bone degradation of crew members and to create new protective designs for space capsules in the future.