Further analyses of human kidney cell populations separated on the Space Shuttle.

Cultured human embryonic kidney cells were separated into electrophoretic subpopulations in laboratory experiments and in two separation experiments on the STS-8 (Challenger) Space Shuttle flight using the mid-deck Continuous Flow Electrophoretic Separator (CFES). Populations of cells from each fraction were cultured for the lifetime of the cells, and supernatant medium was withdrawn and replaced at 4-day intervals. Withdrawn medium was frozen at -120 degrees C for subsequent analysis. Enzyme assays, antibodies and gel electrophoresis were used as analytical tools for the detection and quantitation of plasminogen activators in these samples. These assays of frozen culture supernatant fluids confirmed the electrophoretic separation of plasminogen-activator producing cells from non-producing cells, the isolation of cells capable of sustained production, and the separation of cells that produce different plasminogen activators from one another.     

Arthropod model systems for studying complex biological processes in the space environment.

Three arthropod systems are discussed in relation to their complementary and potential use in Space Biology. In a next biosatellite flight, Drosophila melanogaster pre-adapted during several months to different g levels will be flown in an automatic device that separates parental from first and second generations. In the same flight, flies will be exposed to microgravity conditions in an automatic unit in which fly motility can be recorded. In the International Microgravity Laboratory-2, several groups of Drosophila embryos will be grown in Space and the motility of a male fly population will be video-recorded. In the Biopan, an ESA exobiology facility that can be flown attached to the exterior of a Russian biosatellite, Artemia dormant gastrulae will be exposed to the space environment in the exterior of the satellite under a normal atmosphere or in the void. Gastrulae will be separated in hit and non-hit populations. The developmental and aging response of these animals will be studied upon recovery. With these experiments we will be able to establish whether exposure to the space environment influences arthropod development and aging, and elaborate on some of the cellular mechanisms involved which should be tested in future experiments.     

How gravity acts on Paramecium: new insights from free-fall experiments.

The swimming behaviour of Paramecium is affected by media of various specific gravities. At 1g, the negative gravitaxis of Paramecium virtually disappears in solutions the specific gravity of which is about the same as that of the organism (1.04). In solutions with a higher specific gravity (1.08), Paramecium becomes positively gravitactic. We recorded the swimming tracks of Paramecium in these media on videotape before, during and after free-falls. The records show that the density-dependent differences in the swimming behaviour disappeared immediately following the onset of the free-fall. The recorded tracks and distributions of cells in the experimental chambers were compared with computer-simulated traces and distributions based on gravitactic and gravikinetic models proposed for Paramecium. Our preliminary analysis favors a novel gravitactic mechanism involving modification of the ciliary movement The drop shaft at the Japan Microgravity Center, Hokkaido (JAMIC) was used for the free-fall experiments.     

Simulated microgravity does not alter epithelial cell adhesion to matrix and other molecules.

Microgravity has advantages for the cultivation of tissues with high fidelity; however, tissue formation requires cellular recognition and adhesion. We tested the hypothesis that simulated microgravity does not affect cell adhesion. Human colorectal carcinoma cells were cultured in the NASA Rotating Wall Vessel (RWV) under low shear stress with randomization of the gravity vector that simulates microgravity. After 6-7 days, cells were assayed for binding to various substrates and compared to cells grown in standard tissue culture flasks and static suspension cultures. The RWV cultures bound as well to basement membrane proteins and to CEA, an intercellular adhesion molecule, as control cultures did. Thus, microgravity does not alter epithelial cell adhesion and may be useful for tissue engineering.     

Drop motion in a rotating immiscible liquid body

The interaction of liquids is an important part of many processes and is often used for such purposes as causing a separation of solution components by mass transfer between phases, allowing a chemical reaction between liquids or their components, providing direct contact heating or cooling, or creating emulsions. Containerless space processing may well involve the interaction of liquids in the form of drops. For example it may be desirable to form an alloy in space by merger of two or more drops. Encapsulation of a volatile liquid by a second less volatile and immiscible liquid might be a means for avoiding volatilization. Rotation of such systems would enhance mixing where desirable and/or provide means for drop management. In this study, single drop behavior in rotating liquid bodies is studied experimentally.     

Short-term responses of gravitaxis to altered gravity in Paramecium.

Negative gravitaxis of Paramecium almost disappeared in solutions having specific gravity about the same as that of the organisms (1.04). The taxis turned to positive in solutions of specific gravity 1.08. Using a drop shaft at the Japan Microgravity Center, Hokkaido (JAMIC) we examined how swimming behaviour in these media was modified by changing gravitational conditions before, during and after free-fall. Tracks of swimming cells recorded on videotape indicate that the swimming cells continued upward and downward shift depending on the specific gravity of the external medium under 1-g conditions and these vertical displacements disappeared immediately after the moment of launch. The effectiveness of changing gravity to induce displacement of the cells seems to depend on the orientation of the cells to gravity. These results suggest a corelation between vertical displacement of the cell through the medium and a gravitactic mechanism in Paramecium.     

电子对撞机束流管冷却系统模糊控制仿真

电子对撞机运行过程中,会产生很多同步辐射热,这些热量均通过束流管冷却系统散失出去.针对束流管冷却这个关键技术难题,研制了一种全新的基于模块化的主动式液体冷却系统;通过合理的分析与简化,应用集总参数法建立了束流管冷却系统的温度动态特性模型,导出了并联调节阀流量特性的数学方程;根据模型给出了相应的模糊控制策略,并在此基础上对束流管冷却系统的控制策略进行了仿真研究.仿真结果表明:采用模糊控制策略的冷却系统具有稳定、超调小的特点,优于传统的比例积分微分(PID)控制策略,能很好地满足电子对撞机的要求,并可以据此指导工程设计.     

Space environmental factors affecting responses to radiation at the cellular level.

Previous space experiments suggest a high value for the RBE of cosmic radiation. A possible explanation could be a change in cell radiosensitivity due to a combined effect of radiation and other factors related to the space environment and to the space flight. Results of the EXOBLOC II experiment support this assumption. On earth, vibrations or accelerations applied before or after irradiation can change the responses to radiation. Microgravity could be the main factor affecting the radiosensitivity and DNA repair but this hypothesis must be confirmed by additional experiments.     

Simulation of launch and re-entry acceleration profiles for testing of shuttle and unmanned microgravity research payloads

Microgravity experiments designed for execution in Get-Away Special canisters, Hitchhiker modules, and Reusable Re-entry Satellites will be subjected to launch and re-entry accelerations. Crew-dependent provisions for preventing acceleration damage to equipment or products will not be available for these payloads during flight; therefore, the effects of launch and re-entry accelerations on all aspects of such payloads must be evaluated prior to flight. A procedure was developed for conveniently simulating the launch and re-entry acceleration profiles of the Space Shuttle (3.3 and 1.7 × g maximum, respectively) and of two versions of NASA's proposed materials research Re-usable Re-entry Satellite (8 × g maximum in one case and 4 × g in the other). By using the 7 m centrifuge of the Gravitational Plant Physiology Laboratory in Philadelphia it was found possible to simulate the time dependence of these 5 different acceleration episodes for payload masses up to 59 kg. A commercial low-cost payload device, the “Materials Dispersion Apparatus” of Instrumentation Technology Associates was tested for (1) integrity of mechanical function, (2) retention of fluid in its compartments, and (3) integrity of products under simulated re-entry g-loads. In particular, the sharp rise from 1 g to maximum g-loading that occurs during re-entry in various unmanned vehicles was successfully simulated, conditions were established for reliable functioning of the MDA, and crystals of 5 proteins suspended in compartments filled with mother liquor were subjected to this acceleration load.     

模拟微重力对人骨髓间充质干细胞向成骨细胞分化中细胞信号通路影响的分析

通过模拟来研究微重力对hMSC向成骨细胞分化的影响,并利用相关信号通路的激活剂或抑制剂来调节这一分化过程.研究结果表明,在成骨细胞分化诱导条件下,微重力降低了hMSC向成骨细胞定向分化的能力,并且成骨细胞标记性基因的表达明显降低, Runt相关转录因子2(Ruax2)的表达受到抑制.相反,过氧化物酶体增殖激活受体γ(PPARγ2)的表达则增加.同时,微重力也降低了ERK的磷酸化水平,而增加了p38MAPK的磷酸化水平.使用p38MAPK的抑制剂SB203580能够抑制p38MAPK的磷酸化,但不能降低PPARγ2的表达水平.骨形态发生蛋白(BMP)能增加Runx2的表达水平.成纤维细胞生长因子2(FGF2)增加了ERK的磷酸化水平,但也不能显著增加成骨细胞标记性基因的表达水平.采用BMP,FGF2和SB203580三种因子组合来调控微重力下的成骨细胞分化能力,结果表明三者的协同作用能显著逆转微重力对成骨细胞定向分化的生物学效应.研究结果还说明,模拟微重力应该是通过不同的细胞信号通路来抑制成骨细胞分化和提升脂肪细胞分化的.      

Properties of electrophoretic fractions of human embryonic kidney cells separated on Space Shuttle flight STS-8.

Suspensions of cultured primary human embryonic kidney cells were subjected to continuous flow electrophoresis on Space Shuttle flight STS-8. The objectives of the experiments were to obtain electrophoretically separated fractions of the original cell populations and to test these fractions for the amount and kind of urokinase (a kidney plasminogen activator that is used medically for digesting blood clots), the morphologies of cells in the individual fractions, and their cellular electrophoretic mobilities after separation and subsequent proliferation. Individual fractions were successfully cultured after return from orbit, and they were found to differ substantially from one another and from the starting sample with respect to all of these properties.     

Comparative characteristic of mitochondria ultrastructural organization in Chlorella cells under altered gravity conditions.

Results from experiments that used cells from the unicellular alga Chlorella vulgaris (strain Larg-1) grown on a clinostat, demonstrated the occurrence of rearrangements in cellular organelles, including changes in the mitochondrial ultrastructure compared to controls. Changes in mitochondrial structure were observed in auto- and heterotrophic regimes of cells grown in altered gravity conditions, especially in long-term experiments. The mitochondrial rearrangements become apparent during cell proliferation, which resulted in an increase in the relative volume of mitochondria per cell: up to 2.7 +/- 0.3% in short-term clino-rotation (2.2 +/- 0.1% in the control) and up to 5.3 +/- 0.4% and 5.1 +/- 0.4% in long-term clinorotation (2.3 +/- 0.2% in the control). The size of the mitochondria and their cristae increased in cells grown under long-time clinorotation. In addition, hypertrophied organelles, not typical for this strain, were observed. These changes in the cells were accompanied by increased electron density of the matrix and a well-ordered topography of the cristae. To examine the separation of oxidative phosphorylation and respiration, an inhibitory agent 2,4-dinitrophenol (2,4-DNP) was applied to cells which resulted in insignificant volume changes of the mitochondria (2.5 +/- 0.4% versus 2.1 +/- 0.2% in the control). The increase of mitochondrial size with regularly arranged cristae, with more condensed matrix and extension of cristae areas of clino-rotated cells, may demonstrate higher functional activity of the mitochondria under altered gravity conditions. Changes observed early in clinorotated cells, in particular the increased level of respiration, adenylate content (especially ATP) and more intensive electron-cytochemical reactions of Mg2(+)-ATPase and succinate [correction of succinat] dehydrogenase (SDH) in mitochondria (including hypertrophic organelles), also suggest increased activity of mitochondria from cells grown under altered gravity conditions compared to controls.     

Investigations on temperature distribution of satellite surfaces affected by solar absorptivity

Based on the Monte Carlo ray-tracing method, the network coefficients of thermal network model describing the radiation heat transfer among satellite surfaces is solved by considering the surface material optical characters. It is superiority to the conventional Gebhart’s method in view of the grey body and the diffuse reflection assumptions. The zone leveling method is used to discrete the governing equations and the solar absorpivity is separated and considered to be an important correction parameter. Effects of the solar incidence round angle, the zenith angle and the ratio of absorpivity to emissivity (RAE) on temperature distribution are numerically simulated and discussed in detail. The higher or the lower the RAE may be lead to the alternative heating and cooling tend with a larger heating or cooling velocity of main body surfaces than the solar array surfaces. Furthermore, maximum temperature of main body is almost larger than solar arrays. Under the same RAE, solar incidence angle make a great effect on the uniform character of temperature distribution.     

一种富氧中空纤维膜组件的温度特性

通过实验研究温度对中空纤维膜组件氧/氮分离性能的影响,结果表明温度能够明显影响膜的分离特性,并且不影响富氧浓度,可以通过控制膜组件的使用温度来增加富氧产量.      

失重飞机上进行的电泳分离实验

介绍了用A3—l连续自由流电泳仪在俄罗斯失重飞机上进行的实验情况及结果．进行了4个架次的飞行试验，获得了飞行试验的数据和电泳分离图像．试验结果表明，电泳仪装置工作良好，从分离图像可以看出重力变化对电泳分离过程的影响并证实微重力环境对生物材料的电泳分离是有利的．     

Formation of otoconia in the Japanese red-bellied newt, Cynops pyrrhogaster.

Pre-mated adult female newts and fertilized eggs will be flown on the International Microgravity Laboratory-2 flight, in 1994. One objective of the flight will be to observe the influence of microgravity on the development of the gravity-sensing organs in the inner ear. These organs contain sensory hair cells covered by a layer of dense stones (otoconia). Gravity and linear acceleration exert forces on these masses, leading to excitation of the nerve fibers innervating the hair cells. If the production of the otoliths is regulated to reach an optimal weight, their development might be abnormal in microgravity. Ground-based control experiments are reported describing the developmental sequence in which both the otoliths and their associated sensory epithelium and the semicircular canals appear and develop. Three-dimensional reconstruction of serial sections through the otic vesicle of newt embryos at stages 31 through 58 demonstrate the first appearance, relative position and growth of the otoliths. Reports of experiments in which fertilized frog eggs were flown on a Russian Cosmos mission conclude that the utricular otolith is increased in volume, whereas the saccular otolith maintains normal size, suggesting that at least in the utricle, the weight of the otolith might be regulated.     

Production of hydrocarbons and nitriles by electrical processes in Titan's atmosphere.

Although lightning has not been observed in Titan's atmosphere, the presence of methane rain in the troposphere suggests the possibility of electrical activity in the form of corona and/or lightning discharges. Here we examine the chemical effects of these electrical processes on a Titan simulated atmosphere composed of CH4 in N2 at various mixing ratios. Corona discharges were simulated in two different experimental arrays. For the detection of reactive intermediates we used a mass spectrometer to study the main positive ions arising by bombarding low-energy electrons from a hot filament into low-pressure methane. The final stable products, generated by applying a high voltage in a coaxial reactor with either positive or negative polarity, were separated and detected by gas chromatography-Fourier transform infrared spectroscopy and electron impact mass spectrometry (GC-FTIR-MS). Lightning discharges were simulated by a hot and dense plasma generated by a Nd-YAG laser and the final products were separated and detected by GC-FTIR-MS. Corona discharges produce linear and branched hydrocarbons as well as nitriles whereas lightning discharges generate mainly unsaturated hydrocarbons and nitriles. Lightning discharges are about 2 orders of magnitude more efficient in product formation than corona discharges.     

The role of gravity in the evolutionary emergence of multicellular complexity: Microgravity effects on arthropod development and aging

While experiments carried out in Space with isolated cells have shown that eucaryotic cells are able to sense and respond to the absence of gravity by modifying their reactions, experiments in which more complex processes have been investigated, such as Biological Systems undergoing development under Microgravity, have been surprisingly unaffected by the space environment. This can be considered a curious result since all organisms are evolutionarily adapted to the current level of the gravity force in our planet and should eventually change if this parameter will vary in a permanent manner. In fact, the small effects of the modifications in gravity on development in short term experiments may be equivalent to the difficulties in detecting the involvement of other basic physical processes such as diffusion-controled auto-organizative reactions in currently developing biological systems. An apparent exception to this lack of effect is experiments where brine shrimp dormant gastrulae directly exposed to the space environment accumulate developmental defects as a consequence of cosmic irradiation. In this article we discuss the idea that at a certain stage during the evolutionary emergence of multicellular organisms the cues laid by generic forces such as gravity were involved in the evolutionary organization of these primitive organisms. As evolution proceed, these early mechanisms may have been obscured and/or made redundant by the appearance of new internal, environment-independent biological regulatory mechanisms. On the other hand, behavioral responses that may be important, for example, in setting the life-spans of organisms may still be more readily susceptible to manipulation by external cues as experiments carried out by our group in Space and on the ground with Drosophila melanogaster indicate.     

Gravitational influence on binary alloy melt equilibria

Micro- and macrosegregation, the nonuniform redistribution of solute during solidification, are common to both casting and welding, processes of fundamental importance in materials engineering. In multicomponent crystal growth where solid/liquid density differences are appreciable gravity-induced separation can lead to significant spatial variations in resulting ingot composition. In fact, this phenomenon is also operative in liquid/liquid systems such as monotectic alloys exhibiting a liquid miscibility gap, where buoyancy-driven flows can result in a sometimes unwanted separation of phases upon cooling through the miscibility dome.     

Focal contacts organization in osteoblastic cells under microgravity and cyclic deformation conditions.

We compared quantitatively vinculin-related adhesion parameters in osteoblastic cells submitted to opposite mechanical stresses, i.e., low deformation and frequency strain regimens (stretch condition) and microgravity exposure (relaxed condition). Cyclic deformation induced a biphasic response comprising new focal contacts formation followed by their clustering in ROS cells. Microgravity exposure induced a reduction in focal contact number and clustering in ROS cells. We previously demonstrated that 1% cyclic deformations at 0.05 Hz during a daily 10 min episode over 7 days stimulated ROS 17/2.8 growth as compared to static culture whereas relaxed ROS proliferated similarly to static culture (BC). To evaluate whether the proliferation (stretch) or the survival (relaxed) status of ROS cells influences focal contact organization, we inhibited ERKs proliferative-dependent pathway. Inhibition of proliferation by PD98059 was overcome although not fully restored by stretch. Furthermore stretch-induced clustering of vinculin-positive contacts still occurs in the presence of ERKs inhibitor, whereas the increase in focal contact number is abolished. In conclusion, we showed that focal contacts are mechanoeffectors and that hyper-mechanical stimulation could up regulate focal contacts size as compared to hypo-mechanical that down regulate clusterization.