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Abstract: The bone loss induced by microgravity is partly due to the decrease of mature osteoblasts. In the present study, we employed the random positioning machine (RPM) to simulate microgravity and investigated the acute effects of simulated microgravity on the differentiation of 2T3 preosteoblasts. Following 7days’ culture under normal (1 g) condition, cells were exposed to simulated microgravity for 24h. The results showed that 24h treatment of simulated microgravity significantly decreased alkaline phosphatase (ALP) activity without changing the cell morphology. In addition, the mRNA expressions of osteogenic genes, including runt-related gene 2 (Runx2), osterix, osteocalcin (OC), type I collagen (Col I) and bone morphogenetic protein (BMP), were dramatically downregulated. Moreover, western blot analysis of total extracellular signal-regulated kinase (Erk) and phosphorylated Erk (p-Erk) indicated that p-Erk level, which represents the Erk activation status, was increased. Taken together, our results suggested that acute exposure to simulated microgravity inhibited osteoblast differentiation through modulating the expression of osteogenic genes and the Erk activity. These findings provide new insight for bone loss due to microgravity and unloading. [Copyright &y& Elsevier]

Abstract: Arabidopsis gene expression microarray (44K) was used to detect genes highly induced under simulated microgravity stress (SMS). Ten SMS-inducible genes were selected from the microarray data and these 10 genes were found to be abundantly expressed in 3-week-old plants. Nine out of the 10 SMS-inducible genes were also expressed in response to the three abiotic stresses of drought, touch, and wounding in 3-week-old Arabidopsis plants respectively. However, WRKY46 was elevated only in response to SMS. Six other WRKY genes did not respond to SMS. To clarify the characteristics of the genes expressed at high levels in response to SMS, 20 cis-elements in the promoters of the 40 selected genes including the 10 SMS-inducible genes, the 6 WRKY genes, and abiotic stress-inducible genes were analyzed and their spatial positions on each promoter were determined. Four cis-elements (M/T-G-T-P from MYB1AT or TATABOX5, GT1CONSENSUS, TATABOX5, and POLASIG1) showed a unique spatial arrangement in most SMS-inducible genes including WRKY46. Therefore the M/T-G-T-P cis-element patterns identified in the promoter of WRKY46 may play important roles in regulating gene expression in response to SMS. The presences of the cis-element patterns suggest that the order or spatial positioning of certain groups of cis-elements is more important than the existence or numbers of specific cis-elements. Taken together, our data indicate that WRKY46 is a novel SMS inducible transcription factor and the unique spatial arrangement of cis-elements shown in WRKY46 promoter may play an important role for its response to SMS. [Copyright &y& Elsevier]

Abstract: Hematopoietic progenitor cell proliferation can be alternated on either spaceflight or under simulated microgravity experiments on the ground; however, the underlying mechanism remains largely unknown. In the present study, we have demonstrated that exposure of human erythropoietin (EPO)-dependent leukemia cell line UT-7/EPO cells to conditions of simulated microgravity with a rotary culture instrument significantly inhibited the cellular proliferation rate. Adding higher concentrations of EPO to the culture medium failed to improve the inhibitory status. Cell apoptosis was detected by fluorescence staining of cell nuclei and a flow cytometry assay using Annexin V/PI double staining. This microgravity-induced apoptosis in UT-7/EPO cells could be blocked by a pancaspase inhibitor Z-VAD-FMK. Immunoblotting demonstrated that rotary culture resulted in a reduction of the expression of Bcl-xL, an anti-apoptotic protein, and the cleavage of caspase-3. Furthermore, rotary culture reduced surface localization and protein content, as well as the mRNA expression of erythropoietin receptor (EPOR) of UT-7/EPO. Take together, the findings indicated that simulated microgravity may induce mitochondrial related apoptosis of UT-7/EPO cell through depressing the EPO–EPOR pathway. [ABSTRACT FROM AUTHOR]

Abstract: This study investigated the time-course of stress and recovery states and their relations to social support and personality traits in healthy women during a long-term head-down tilt bed rest. Personality, social support and affective states were assessed in 16 women exposed to simulated microgravity for a 60-day duration involving three stages: a 20-day baseline control period (BDC), a 60-day head-down tilt bed rest (HDT) and a 20-day post-HDT ambulatory recovery period (R+). Participants were divided into two groups: an exercise (Exe, n =8) and a control group (Ctl, n =8). All the participants experienced significantly more stress during the HDT period. But exercise did not improve the impaired effects of simulated microgravity. The Exe group perceived more stress and less recovery than the Ctl group during the HDT period. Among the five major personality factors, only Neuroticism was related to both social and affective variables. Neuroticism was positively associated with stress and negatively associated with recovery and social support (S-SSQ). Practical implications in psychological countermeasures for better dealing with the key human factor in spaceflights are discussed. [Copyright &y& Elsevier]

Abstract: 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 15rpm). After culture for 24h, 48h, 72h, and 96h, 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 12h to 96h). 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 12h. But 12h later (from 24h to 60h), 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 12h, including a decrease in cyclin A, and the increasing in cyclin B, D1 and E, and then (from 24h to 36h) began to restore to control levels. After RCCS culture for 12–36h, 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. [Copyright &y& Elsevier]

Abstract: The first experiments using machines and instruments to manipulate gravity and thus learn about its impact to this force onto living systems were performed by Sir Thomas Andrew Knight in 1806, exactly two centuries ago. What have we learned from these experiments and in particular what have we learned about the use of instruments to reveal the impact of gravity and rotation on plants and other living systems? In this essay I want to go into the use of instruments in gravity related research with emphases on the Random Positioning Machine, RPM. Going from water wheel via clinostat to RPM, we will address the usefulness and possible working principles of these hypergravity and mostly called microgravity, or better, micro-weight simulation techniques. [Copyright &y& Elsevier]