Analysis of Growth Characteristics and Differentially Expressed Homologous Genes in Rhodobacter sphaeroides under Normal and Simulated Microgravity Conditions

The term “microgravity” is used to describe the “weightlessness” or “zero-g” circumstances that can only be found in space beyond earth’s atmosphere. Rhodobacter sphaeroides is a gram-negative purple phototroph, used as a model organism for this study due to its genomic complexity and metabolic versatility. Its genome has been completely sequenced, and profiles of the differential gene expression under aerobic, semi-aerobic, and photosynthetic conditions were examined. In this study, we hypothesized that R. sphaeroides will show altered growth characteristics, morphological properties, and gene expression patterns when grown under simulated microgravity. To test that, we measured the optical density and colony-forming units of cell cultures grown under both microgravity and normal gravity conditions. Differences in the cell morphology were observed using scanning electron microscopy (SEM) images by measuring the length and the surface area of the cells under both conditions. Furthermore, we also identified homologous genes of R. spheroides using the differential gene expression study of Acidovorax under microgravity in our laboratory. Growth kinetics results showed that R. sphaeroides cells grown under microgravity experience a shorter log phase and early stationary phase compared to the cells growing under normal gravity conditions. The length and surface area of the cells under microgravity were significantly higher confirming that bacterial cells experience altered morphological features when grown under microgravity conditions. Differentially expressed homologous gene analysis indicated that genes coding for several COG and GO functions, such as metabolism, signal-transduction, transcription, translation, chemotaxis, and cell motility are differentially expressed to adapt and survive microgravity.

Simulated Microgravity Conditions and Carbon Ion Irradiation Induce Spermatogenic Cell Apoptosis and Sperm DNA Damage

Objective To investigate the effect of simulated microgravity and carbon ion irradiation （CIR） on spermatogenic cell apoptosis and sperm DNA damage to the testis of male Swiss Webster mice, and assess the risk associated with space environment. Methods Sperm DNA damage indicated by DNA fragmentation index （DFI） and high DNA stainability （HDS） was measured by sperm chromatin structure assay （SCSA）. Apoptosis of spermatogenic cells was detected by annexin V-propidium iodide assay. Bax （the expression levels of p53） and proliferating cell nuclear antigen （PCNAI were measured by immunoblotting; p53 and PCNA were located by immunohistology. Results HDS, DFI, apoptosis index, and the expression levels of p53 and Bax were detected to be significantly higher in the experimental groups （P〈0.05） compared with those in the control group, however, the PCNA expression varied to a certain degree, p53- and PCNA- positive expression were detected in each group, mainly in relation to the spermatogonic cells and spermatocytes. Conclusion The findings of the present study demonstrated that simulated microgravity and CIR can induce spermatogenic cell apoptosis and sperm DNA damage. Sperm DNA damage may be one of the underlying mechanisms behind male fertility decline under space environment. These findings may provide a scientific basis for protectint~ astronauts and space traveler＇s health and safety.

Differentiation of Mesenchymal Stem Cells towards a Nucleus Pulposus-like Phenotype Utilizing Simulated Microgravity In Vitro

Mesenchymal stem cells （MSCs） were induced into a nucleus pulposus-like phenotype utilizing simulated microgravity in vitro in order to establish a new cell-based tissue engineering treatment for intervertebral disc degeneration. For induction of a nucleus pulposus-like phenotype, MSCs were cultured in simulated microgravity in a chemically defined medium supplemented with 0 （experimental group） and 10 ng/mL （positive control group） of transforming growth factor β1 （TGF-β1）. MSCs cultured under conventional condition without TGF-β1 served as blank control group. On the day 3 of culture, cellular proliferation was determined by WST-8 assay. Differentiation markers were evaluated by histology and reverse transcriptase-polymerase chain reaction （RT-PCR）. TGF-β1 slightly promoted the proliferation of MSCs. The collagen and proteoglycans were detected in both groups after culture for 7 days. The accumulation of proteoglycans was markedly increased. The RT-PCR revealed that the gene expression of Sox-9, aggrecan and type Ⅱ collagen, which were chondrocyte specific, was increased in MSCs cultured under simulated microgravity for 3 days. The ratio of proteoglycans/collagen in blank control group was 3.4-fold higher than positive control group, which denoted a nucleus pulposus-like phenotype differentiation. Independent, spontaneous differentiation of MSCs towards a nucleus pulposus-like phenotype in simulated microgravity occurred without addition of any external bioactive stimulators, namely factors from TGF-β family, which were previously considered necessary.

Effect of Simulated Microgravity and its Associated Mechanism on Pulmonary Circulation in Rats

Objective To study the effect of Simulated Microgravity and its Associated Mechanism on Pulmonary Circulation in Rats. Methods Rat tail-suspension model was used to simulate the physiological effects of microgravity and changes in pulmonary blood vessel morphology, pulmonary arterial and venous blood pressure, pulmonary vascular resistance, pulmonary vasomotoricity, as well as the regulation of pulmonary circulation by cytokines produced and released by the lung of rats were measured. Results The walls of pulmonary blood vessels of rats were thickened, and the pulmonary artery was reconstructed with increased pulmonary vascular resistance. The pulmonary blood vessels of rats became more prone to dilation as contractions increased. Rat epithelial Adrenomedulin gene transcription and protein expression were upregulated. The level of basic fibroblast growth Factor of rat was also elevated. Conclusion Findings from the present study on rats revealed that the microgravity can affect pulmonary blood vessel structure, pulmonary arterial pressure, and pulmonary blood vessel self-regulation and cytokine production.

The study on the mechanical characteristics of articular cartilage in simulated microgravity

The microgravity environment of a long-term space flight may induce acute changes in an astronaut＇s musculo-skeletal systems. This study explores the effects of simulated microgravity on the mechanical characteristics of articular cartilage. Six rats underwent tail suspension for 14 days and six additional rats were kept under normal earth gravity as controls. Swelling strains were measured using high-frequency ultrasound in all cartilage samples subject to osmotic loading. Site-specific swelling strain data were used in a triphasic theoretical model of cartilage swelling to determine the uniaxial modulus of the cartilage solid matrix. No severe surface irregularities were found in the cartilage samples obtained from the control or tail-suspended groups. For the tail-suspended group, the thickness of the cartilage at a specified site, as determined by ultrasound echo, showed a minor decrease. The uniaxial modulus of articular cartilage at the specified site decreased significantly, from （6.31 ± 3.37） MPa to （5.05 ± 2.98）MPa （p 〈 0.05）. The histology- stained image of a cartilage sample also showed a reduced number of chondrocytes and decreased degree of matrix staining. These results demonstrated that the 14 d simulated microgravity induced significant effects on the mechanical characteristics of articular cartilage. This study is the first attempt to explore the effects of simulated microgravity on the mechanical characteristics of articular cartilage using an osmotic loading method and a triphasic model. The conclusions may provide reference information for manned space flights and a better understanding of the effects of microgravity on the skeletal system.

Effects of simulated microgravity on antioxidant system in testis of adult rats

Objective： To study the effects of simulated microgravity induced by tail-suspension on histological structures and antioxidant system in testis of adult rats. Methods： Forty Spraque-Dawley adult male rats were randomly divided into four groups, two experimental groups and two synchronic control groups. Rats in the two experimental groups were tail-suspended for 14 d and 28 d respectively, then serum hormone, testicular morphology and biochemical changes were evaluated. Results： Compared with the synchronic controls, serum testosterone （T） levels and testicular superoxide dismutase （SOD） significantly decreased（P 〈 0.05）, while testicular malondialdehyde（MDA） significantly increased （P 〈 0.05） in tail-suspended rats. The changes were more prominent in the 28 d tail-suspended rats. In addition, hematoxylin and eosin （HE） staining showed that seminiferous tubules atrophied, spermatogenic cells decreased, and seminiferous epithelial cells disintegrated and shed, abnormal multinucleated giant cells appeared and the lumina were azoospermic in tail-suspended rats. ConcIusion： Simulated microgravity induced by tailsuspension has a harmful effect on male reproduction of adult rats by decreasing the ability of testicular antioxidant defense.

Protective effects of Dragon's Blood on blood coagulation and NO/iNOS level in myocardium and serum of rats in simulated microgravity

To investigate effects of Dragon＇s Blood（DB）,a traditional Chinese medicine,on blood coagulation and NO/iNOS level in myocardium and serum of rats in simulated microgravity for the first time,Sprague Dawley（SD）rats were randomly divided into six groups：（a）5-day control group,（b）5-day model group,（c）5-day drug group,（d）21-day control group,（e）21-day model group,and（f）21-day drug group.Blood coagulation and NO/iNOS level in myocardium and serum were examined after 5 and 21 days of simulated microgravity respectively.The results showed that blood of tail-suspended rats was in a hypercoagulable state that could not be converted with time extending.Conversely,DB changed these parameters towards normal level and the curative effects became better when tail-suspension lasted till the 21 st day.NO concentration of both myocardium and serum for two periods all increased markedly and DB could effectively reduce these increases except that of 21-day myocardium NO.Activity of iNOS increased markedly as early as 5 days and became more serious on the 21 st day,while DB showed preventive effect on the 21 st day.Western Blot analysis illustrated that the expression of iNOS in the 5-day model group increased significantly over the 5-day control group and the expression in the 5-day drug group dramatically returned to the normal level.The similar trend was observed on the 21-day groups without notable variances.The findings of this study can serve for the further use of Dragon＇s Blood in space diseases.

Protective Effect of Space Ginseng Yeast on Human Primary Fibroblasts under Simulated Microgravity Condition

[Objectives] To study the protective effect of Space Ginseng Yeast on human primary fibroblasts. [Methods] Multiple indicators were used to simulate the microgravity effect,so as to study the protective effect of Space Ginseng Yeast on human primary fibroblasts under simulated microgravity condition. [Results]Space Ginseng Yeast could improve changes of cell morphology and decline of the survival rate caused by microgravity,restore the expression and effective arrangement of cytoskeletal protein Tubulin,extracellular matrix proteins Integrin and Fibronectin,and can inhibit the abnormal expression of MMP-1. The protective effect of Space Ginseng Yeast diluted 20-500 times is better,and the protective effect of Space Ginseng Yeast diluted 100 times is the most obvious. [Conclusions] Space Ginseng Yeast can protect damage of human primary fibroblasts caused by simulated microgravity.

THE ROLE OF MUSCLE SPINDLE IN MUSCLE ATROPHY INDUCED BY SIMULATED MICROGRAVITY

Objective To compare the cross section area (CSA) and the immunoreactivity of conjugated ubiquitin in soleus extrafusal and intrafusal fibers after simulated microgravity and to demonstrate the role of muscle spindle in muscle atrophy induced by simulated microgravity. Methods The immunohistochemical technique (ABC) and image analysis were used to assess the conjugated ubiquitin immunostaining and the cross sectional area of intrafusal and extrafusal fibers of soleus in simulated microgravity rats. Results ①Tail suspension caused a progressive loss of soleus mass. Mean fiber CSA of extrafusal fibers were (7±2)%, (21±4)% and (32±7)% smaller after 3 days, 7 days and 14 days suspension, respectively. While the CAS of intrafusal fibers (bag + chain fibers) were (14±3)% ( P < 0.05 ), (30±7)% ( P < 0.01 ) and (44±10)% ( P < 0.01 ) smaller after 3 days, 7 days and 14 days suspension. ② The immunoreactivity of conjugated ubiquitin both in extrafusal and intrafusal fibers increased after tail suspension. The immunoreactivity of intrafusal fibers increased 1 day after suspension and reached the hightest level at 3 days after tail suspension. The immunoreactivity of extrafusal fibers increased after 3 days suspension and reached the highest level after 7 days tail suspension, which was lower than that in intrafusal fibers after 3 days tail suspension. Conclusion These results suggest that soleus atrophy of intrafusal fibers caused by tail suspension is earlier and more severe than that in extrafusal fibers.

Reactive mesoporous silica nanoparticles loaded with limonene for improving physical and mental health of mice at simulated microgravity condition

Astronauts are under high stress for a long time because of the microgravity condition,which leads to anxiety,affects their learning and memory abilities,and seriously impairs the health of astronauts.Aromatherapy can improve the physical and mental health of astronauts in a way that moisturizes them softly and silently.However,the strong volatility of fragrances and inconvenience of aroma treatment greatly limit their application in the field of spaceflight.In this study,reactive mesoporous silica nanoparticles were prepared to encapsulate and slowly release limonene.The limonene loaded nanoparticles were named limonene@mesoporous silica nanoparticles-cyanuric chloride(LE@MSNs-CYC).LE@MSNs-CYC were then applied to wallpaper to improve the convenience of aromatherapy.LE@MSNs-CYC could chemically react with the wallpaper,thus firmly adsorbed on the wallpaper.In the following,the mice were treated with hindlimb unloading(HU)to simulate a microgravity environment.The results showed that 28-day HU led to an increase in the level of anxiety and declines in learning,memory,and physical health in mice.LE@MSNs-CYC showed significant relief effects on anxiety,learning,memory,and physical health of HU treated mice.Subsequently,the molecular mechanisms were explored by hypothalamic-pituitary-adrenal axis related hormones,immune-related cytokines,learning,and memory-related neurotransmitters and proteins.

Early changes to the extracellular space in the hippocampus under simulated microgravity conditions

The smooth transportation of substances through the brain extracellular space(ECS)is crucial to maintaining brain function;however,the way this occurs under simulated microgravity remains unclear.In this study,tracer-based magnetic resonance imaging(MRI)and DECS-mapping techniques were used to image the drainage of brain interstitial fluid(ISF)from the ECS of the hippocampus in a tail-suspended hindlimb-unloading rat model at day 3(HU-3)and 7(HU-7).The results indicated that drainage of the ISF was accelerated in the HU-3 group but slowed markedly in the HU-7 group.The tortuosity of the ECS decreased in the HU-3 group but increased in the HU-7 group,while the volume fraction of the ECS increased in both groups.The diffusion rate within the ECS increased in the HU-3 group and decreased in the HU-7 group.The alterations to ISF drainage and diffusion in the ECS were recoverable in the HU-3 group,but neither parameter was restored in the HU-7 group.Our findings suggest that early changes to the hippocampal ECS and ISF drainage under simulated microgravity can be detected by tracer-based MRI,providing a new perspective for studying microgravity-induced nano-scale structure abnormities and developing neuroprotective approaches involving the brain ECS.

Effects of spaceflight and simulated microgravity on cell sub-microstructure and antioxidant enzyme activity in tomato

Controlled ecological life support systems provide food, air, water, and other basic living resources for crew members on long-duration spaceflight missions. Plants are an important basic requirement of these systems and their biological characteristics in space have very high research value. Based on experiments of spaceflight in Shenzhou 8 spacecraft and simulating microgravity effects on three-dimensional （3-D） clinostat, the biological characteristics of tomato＇s leaf cell sub-microstructure and antioxidant enzyme activities were studied and compared in this work. Results showed that leaf cell sub-microstructure of the tomato samples experiencing spaceflight had more changes than effects, and both peroxidase （POD） and superoxide dismutase （SOD） that of the samples processed by simulated microgravity activities increase obviously in both the environments.

Secondary metabolism in simulated microgravity and space flight

Space flight experiments have suggested that microgravity can affect cellular processes in microorganisms.To simulate the microgravity environment on earth,several models have been developed and applied to examine the effect of microgravity on secondary metabolism.In this paper,studies of effects of space flight on secondary metabolism are exemplified and reviewed along with the advantages and disadvantages of the current models used for simulating microgravity.This discussion is both significant and timely to researchers considering the use of simulated microgravity or space flight to explore effects of weightlessness on secondary metabolism.

The subsequent biological effects of simulated microgravity on endothelial cell growth in HUVECs

Purpose： Microgravity is known to cause endothelium dysfunction in astronauts returning from spaceflight. We aimed to reveal the regulatory mechanism in alterations of human endothelial cells after simulated microgravity （SMG）. Methods： We utilized the rotary cell culture system （RCCS-1） to explore the subsequent effects of SMG on human umbilical vein endothelial cells （HUVECs）. Results： SMG-treated HUVECs appeared obvious growth inhibition after return to normal gravity, which might be attributed to a set of responses including alteration of cytoskeleton, decreased cell adhesion capacity and increased apoptosis. Expression levels of mTOR and its downstream Apaf-1 were increased during subsequent culturing after SMG. miR-22 was up-regulated and its target genes SRF and LAMC1 were down-regulated at mRNA levels. LAMC1 siRNAs reduced cell adhesion rate and inhibited stress fiber formation while SRF siRNAs caused apoptosis. Conclusion： SMG has the subsequent biological effects on HUVECs, resulting in growth inhibition through mTOR signaling and miR-22-mediated mechanism.

Effects of simulated zero gravity on adhesion,cell structure,proliferation,and growth behavior,in glioblastoma multiforme

All life on Earth has evolved under the influence of continuous gravity,and methods have been developed to balance this influence with the biological evolution of organisms at the cellular and system levels.However,when exposed to zero gravity in space,the balance between cell structure and external forces is destroyed,resulting in changes at the cellular level(e.g.,cell morphology,adhesion,viability,apoptosis,etc.),and understanding the molecular mechanism of cell response to zero gravity will help to cope with diseases that rely on mechanical response.Therefore,biological research in space and zero gravity is a unique step in developing the best anti-cancer treatments,which is a great challenge to humanity.In this study,multicellular glioma cancer cells from a brain tumor in a 72-year-old Iraqi patient were subjected to simulated zero gravity for 24 h,and the results showed that most of the cells lost their adhesion,which is considered to be the first step toward cell apoptosis.In addition to the formation of multicellular spheroids,the results also showed that the inhibition rate for cell death was 32%in comparison to the control cells.Moreover,the cells showed a clear change in their cellular morphology and growth behavior.These results give new hope for fighting cancer distinctively,and such a treatment method has no side effects in comparison to traditional chemical and radiological ones.

Effects of Microgravity on Human Physiology and Ground-Based Models for Space Physiological Studies

With the development of manned spaceflight, more and more researches are involved in the area of gravitation physiology. When astronauts are exposed to microgravity, a series of special physiological or pathological changes will occur, which will start self-regulation mechanisms to reduce abnormalities and help the organism to better adapt to microgravity. However, these adaptive changes may also induce degradation or damage to physiological functions. This paper summarizes the physiological effects of microgravity on the human body from the aspects of skeletal and mineral metabolism, muscle structure and function, vestibular functions, cardiovascular function and pulmonary function, as well as expounds some commonly used ground-based space analogies. The paper will provide a reference for further study on the physiological effects of microgravity.

Research on Suspension Gravity Compensation System of Lunar Rover with Magnetic Levitation Servo

In order to overcome the shortcomings of the traditional sling suspension method,such as complex structure of suspension truss,large running resistance,and low precision of position servo system,a gravity compensation method of lunar rover based on the combination of active suspension and active position following of magnetic levitation is proposed,and the overall design is carried out.The dynamic model of the suspension module of microgravity compensation system was established,and the decoupling control between the constant force component and the position servo component was analyzed and verified.The constant tension control was achieved by using hybrid force/position control.The position following control was realized by using fuzzy adaptive PID(proportional⁃integral⁃differential)control.The stable suspension control was realized based on the principle of force balance.The simulation results show that the compensation accuracy of constant tension could reach more than 95%,the position deviation was less than 5 mm,the position deviation angle was less than 0.025°,and the air gap recovered stability within 0.1 s.The gravity compensation system has excellent dynamic performance and can meet the requirements of microgravity simulation experiment of lunar rover.

Space Life Science of China

In the past two years,China’s space life science has made great progress.Space biomedical and life science programs have carried out ground-based research for the first batch of projects,and are preparing to carry out space-based experiments along with the construction of China’s space station.And space life science payload of the space station completed the development of positive samples.Thus,with the development of lunar exploration and Mars exploration projects,astrobiology research has also made a lot of basic achievements.On the basis of summarizing the development of space life science in China,this paper mainly introduces the important progress of payload technology and life science research.