From ROOTS to GRAVI-1: Twenty Five Years for Understanding How Plants Sense Gravity

In the 1970s, when I started to work on gravitropism at the University Pierre and Marie Curie, Paris), it was well known that statocytes contain voluminous amyloplasts (statoliths) that sediment under the influence of gravity. The role of these organelles in gravisensing was strongly disputed. In 1974, I attended a session of a meeting on gravitropism in Würzburg, where I presented results that supported the involvement of statoliths in the perception of gravity. This meeting had a strong impact on my research, since at that time the Council of Europe was looking for people interested in performing experiments in Space. Our first experiment (ROOTS) was carried out in the Biorack Facility (ESA) in the frame of the Spacelab D1 mission (1985). We had a very efficient help from CNES which developed a very fine hardware to grow lentil seedlings and to chemically fix them at the end of the experiment. The results obtained were surprising since we observed that in microgravity the statoliths were located at one pole of the statocyte and not distributed at random as it was expected. The goal of the following experiment (Spacelab IML-1 mission, 1992) was to determine the threshold stimulation time at 1 ×g (created by centrifugation). It was estimated at 25 s. In the frame of the SMM/03 and SMM/06 missions (1996, 1997), we proved that the statoliths are attached to actin filaments by motor proteins (myosin) that make these organelles move in one preferential direction in microgravity. The analysis of gravisensitivity with clinostats incited us to compare gravisensitivity of lentil roots grown in microgravity or on a 1 × g centrifuge (SMM05 mission, 1997). It was found that the latter were less sensitive than the former. We showed that this was due to the fact that the statoliths are not distributed in the same way in both cases (microgravity or 1 × g centrifuge). All these studies led us to propose a mode of gravity sensing by plants in which elements of the cytoskeleton and stretch activated ion channels are involved. The last experiment (GRAVI-1) which has been carried out (in ISS with the EMCS facility, 2007) dealt with the threshold acceleration that is perceived by roots.     

Effect of Hypergravity on Endothelial Cell Function and Gene Expression

It is well known that endothelial cells (ECs), which play a major role in cardiovascular system functioning, are very sensitive to mechanical stimuli. It has been demonstrated that changes in inertial conditions (i.e. microgravity and hypergravity) can affect both phenotypic and genotypic expression in ECs. In this report we describe the effects of hypergravity on ECs isolated from bovine aorta (BAECs). ECs were repeatedly exposed to discontinuous hypergravity conditions (5 × 10 min at 10×g with 10 min at 1×g between sets), simulated in a hyperfuge. Then, cell morphology and metabolism were analyzed by autofluorescence techniques. The phenotypic expression of cytoskeleton constituents (β-actin, vimentin, tubulin), adhesion and survival signals (integrins), mediators of inflammation and angiogenesis was evaluated by immunocytofluorescence. Quantitative PCR (Q-PCR) with Low Density Arrays (LDAs) was used to evaluate modifications in gene expression. After hypergravity exposure, no significant changes were observed in cell morphology and energy metabolism. Cells remained adherent to the substratum, but integrin distribution was modified. Accordingly, the cytoskeletal network reorganized, documenting cell activation. There was a reduction in expression of genes controlling vasoconstriction and inflammation. Proapoptotic signals were downregulated. On the whole, the results documented that hypergravity exposure maintained EC survival and function by activation of adaptive mechanisms.     

Experimental Study on the Ignition Process of Single Coal Particles at Microgravity

In this paper, the first Chinese microgravity (μ-g) experimental study on coal combustion was introduced. An experimental system used to study the ignition process of single coal particles was built up, complying with the requirements of the 3.5 s drop tower in the National Microgravity Laboratory of China (NMLC). High volatile bituminous and lignite coal particles with diameter of 1.5 and 2.0 mm were tested. The ignition and combustion process was recorded by a color CCD and the particle surface temperature before and at the ignition was determined by the RGB colorimetric method. Comparative experiments were conducted at normal gravity (1-g). The experiments revealed that at different gravity levels, the ignition of all tested coal particles commenced in homogeneous phase, while the shape, structure, brightness and development of the flames, as well as the volatile matter release during the ignition process are different. At μ-g, the part of volatile was released as a jet, while such a phenomenon was barely observed at 1-g. Also, after ignition, flames were more spherical, thicker, laminated and dimmer at μ-g. It was confirmed that ignition temperature decreased as the particle size or volatile content increased. However, contradicted to existing experimental results, provided other experimental conditions except gravity level were the same, ignition temperature of coal particles was about 50–80 K lower at μ-g than that at 1-g.     

The Impact of Simulated Microgravity on the Growth of Different Genotypes of the Model Legume Plant <Emphasis Type="Italic">Medicago truncatula</Emphasis>

Simulated microgravity has been a useful tool to help understand plant development in altered gravity conditions. Thirty-one genotypes of the legume plant Medicago truncatula were grown in either simulated microgravity on a rotating clinostat, or in a static, vertical environment. Twenty morphological features were measured and compared between these two gravity treatments. Within-species genotypic variation was a significant predictor of the phenotypic response to gravity treatment in 100% of the measured morphological and growth features. In addition, there was a genotype–environment interaction (G × E) for 45% of the response variables, including shoot relative growth rate (p <?0.0005), median number of roots (p ～ 0.02), and root dry mass (p <?0.005). Our studies demonstrate that genotype does play a significant role in M. truncatula morphology and affects the response of plants to the gravity treatment, influencing both the magnitude and direction of the gravity response. These findings are discussed in the context of improving future studies in plant space biology by controlling for genotypic differences. Thus, manipulation of genotype effects, in combination with M. truncatula’s symbiotic relationships with bacteria and fungi, will be important for optimizing legumes for cultivation on long-term space missions.     

基于HSV量化的幼儿园户外景观色彩设计研究

目的 探究儿童对幼儿园户外景观色彩空间的感知偏好,提出科学有效的儿童活动空间色彩设计方法。方法 选取福州市25个幼儿园户外活动样地为研究对象,从色彩组成和色彩空间构型建立色彩特征指标体系,运用p5.js软件自拟程序将图片转化为HSV空间并进行分割量化,利用ArcGIS10.2、Fragstats4.2软件计算色彩指数,并通过SPSS软件对色彩特征指标进行相关性分析。结果 对儿童感知偏好影响较强的6个色彩指标为:C1色彩均匀度指数>C4对比程度指标>C2色彩数量>C5自然色占比>SHDI斑块多样性指数>FRAC_MN斑块形状平均分维数,C1、C4随偏好度等级呈非线性变化,C2、C5、SHDI、FRAC_MN与偏好度等级成正比;提取出四大色彩特征公因子,分别为色彩均匀度及色彩数量因子F1、色彩对比及色彩饱和度因子F2、色彩斑块分维度因子F3、色彩斑块多样性及自然色因子F4,权重依次为F1>F2>F3>F4。结论 3种高质量色彩空间模式分别具备色彩对比强、自然色丰富、色彩构成形状多样的色彩特征,为幼儿园户外景观色彩质量提升及其他儿童活动空间的色彩设计提供了理论参考。     

Experimental Study of Subcooled Flow Boiling Heat Transfer on a Smooth Surface in Short-Term Microgravity

The flow boiling heat transfer characteristics of subcooled air-dissolved FC-72 on a smooth surface (chip S) were studied in microgravity by utilizing the drop tower facility in Beijing. The heater, with dimensions of 40 × 10 × 0.5 mm³ (length × width × thickness), was combined with two silicon chips with the dimensions of 20 × 10 × 0.5 mm³. High-speed visualization was used to supplement observation in the heat transfer and vapor-liquid two-phase flow characteristics. In the low and moderate heat fluxes region, the flow boiling of chip S at inlet velocity V =?0.5 m/s shows almost the same regulations as that in pool boiling. All the wall temperatures at different positions along the heater in microgravity are slightly lower than that in normal gravity, which indicates slight heat transfer enhancement. However, in the high heat flux region, the pool boiling of chip S shows much evident deterioration of heat transfer compared with that of flow boiling in microgravity. Moreover, the bubbles of flow boiling in microgravity become larger than that in normal gravity due to the lack of buoyancy Although the difference of the void fraction in x-y plain becomes larger with increasing heat flux under different gravity levels, it shows nearly no effect on heat transfer performance except for critical heat flux (CHF). Once the void fraction in y-z plain at the end of the heater equals 1, the vapor blanket will be formed quickly and transmit from downstream to upstream along the heater, and CHF occurs. Thus, the height of channel is an important parameter to determine CHF in microgravity at a fixed velocity. The flow boiling of chip S at inlet velocity V =?0.5 m/s shows higher CHF than that of pool boiling because of the inertia force, and the CHF under microgravity is about 78–92% of that in normal gravity.     

Gravity Induced Absorption Changes in <Emphasis Type="Italic">Phycomyces blakesleanus</Emphasis> and Coleoptiles of <Emphasis Type="Italic">Zea mays</Emphasis> as Measured on the Drop Tower in Bremen (FRG)

Various spectroscopic experiments performed on the AIRBUS ZERO G in the years 2002 to 2007 clearly exhibited optical reflection changes as a result of gravitational changes (GIAC = Gravity Induced Absorption Change) in Phycomyces sporangiophores and corn coleoptiles. GIACs that occurred during flight parabolas in response to hyper- and microgravity were detected by a micro dual wavelength spectrometer (MDWS) for wavelengths pairs in the visible and the near infrared. We assume that GIACs indicate redox-changes of electron transport components such as flavins and cytochromes. Because microgravity on the AIRBUS 300 ZERO G only amounts to moderate values of ±4 ×10^− 2 g, i.e. far below sensitivity of the MDWS, we performed an experiment on the drop tower in Bremen (Germany) that generates microgravity as low as 4 ×10^− 5 g for 4.7 s. We detected small but significant GIACs during the microgravity phase, different in various specimen.     

Gravitactic signal transduction elements in astasia longa investigated during parabolic flights

Euglena gracilis and its close relative Astasia longa show a pronounced negative gravitactic behavior. Many experiments revealed that gravitaxis is most likely mediated by an active physiological mechanism. The goal of the present study was to examine elements in the sensory transduction by means of inhibitors of gravitaxis and the intracellular calcium concentration during short microgravity periods. During the course of six parabolic flights (ESA 31th parabolic flight campaign and DLR 6th parabolic flight campaign) the effects of trifluoperazine (calmodulin inhibitor), caffeine (phosphodiesterase inhibitor) and gadolinium (blocks mechano-sensitive ion channels) was investigated. Due to the extreme parabolic flight maneuvers of the aircraft alternating phases of 1.8×g_n (about 20 s) and microgravity (about 22 s) were achieved (g_n: acceleration of Earth’s gravity field). The duration of the microgravity periods was sufficient to detect a loss of cell orientation in the samples. In the presence of gadolinium impaired gravitaxis was found during acceleration, while caffeine-treated cells showed, compared to the controls, a very precise gravitaxis and faster reorientation in the 1.8×g_n period following microgravity. A transient increase of the intracellular calcium upon increased acceleration was detected also in inhibitor-treated samples. Additionally, it was found that the cells showed a higher calcium signal when they deviated from the vertical swimming direction. In the presence of trifluoperazine a slightly higher general calcium signal was detected compared to untreated controls, while gadolinium was found to decrease the intracellular calcium concentration. In the presence of caffeine no clear changes of intracellular calcium were detected compared to the control. Dedicated to the memory of our colleague and friend Helmut Wagner     

基于PCCS色彩体系和灰色关联分析的产品色彩设计方法

目的 旨在帮助设计人员把握产品色彩属性与用户意象感知间潜在联系,提高产品色彩设计效率。方法 采用多维统计及聚类分析获取PCCS色彩体系下的代表性色彩样本与语汇样本,并设置色彩语义量化实验获取色彩在各语汇尺度上的意象评值。以获取的评值量化数据为基础结合灰色关联分析,通过色彩转置在Lab均匀色彩空间中建立产品的色彩设计评价模型。结果 以双色儿童玩具车为对象进行产品色彩设计实践,构建的色彩设计评价模型良好达成了产品色彩方案的用色筛选、确认及评价优选等目标。结论 结合灰色关联分析,在PCCS色彩体系下构建的产品色彩设计评价模型能够有效提升产品方案的色彩设计效率,并对产品色彩方案的优选给出直观准确的参考标准。     

新一代载人飞船卫生区功能性空间优化设计研究

目的 开展中国新一代载人飞船卫生区的空间优化研究,探索功能性空间优化设计方法,提高载人飞船空间利用效率,同时提升航天员的舒适性体验。方法 以中国空间站卫生区的功能和结构为依据,探讨中国新一代载人飞船卫生区功能组件的构成;依据航天员在微重力状态下使用卫生区的动作与姿态,建立卫生区空间布置的初步概念;根据卫生区的舱壁布局、微重力下航天员视野区域及最佳操作空间分析,开展相关功能组件建模,基于卫生区的人机交互节点,利用JACK软件进行仿真分析与评估,结合评估结果改进设计方案,并通过追踪航天员活动轨迹产生的包络体完成卫生区的空间优化。结论 通过对比中国新一代载人飞船卫生区优化前后设计方案的舒适性、可操作性和空间尺寸,验证了功能性空间优化设计方法的有效性和可靠性,为载人航天器舒适性与集成化设计提供了参考。     

Relation Between Motility, Accelerated Aging and Gene Expression in Selected Drosophila Strains under Hypergravity Conditions

Motility and aging in Drosophila have proven to be highly modified under altered gravity conditions (both in space and ground simulation facilities). In order to find out how closely connected they are, five strains with altered geotactic response or survival rates were selected and exposed to an altered gravity environment of 2g. By analysing the different motile and behavioural patterns and the median survival rates, we show that altered gravity leads to changes in motility, which will have a negative impact on the flies’ survival. Previous results show a differential gene expression between sessile samples and adults and confirm that environmentally-conditioned behavioural patterns constrain flies’ gene expression and life span. Therefore, hypergravity is considered an environmental stress factor and strains that do not respond to this new environment experience an increment in motility, which is the major cause for the observed increased mortality also under microgravity conditions. The neutral-geotaxis selected strain (strain M) showed the most severe phenotype, unable to respond to variations in the gravitational field. Alternatively, the opposite phenotype was observed in positive-geotaxis and long-life selected flies (strains B and L, respectively), suggesting that these populations are less sensitive to alterations in the gravitational load. We conclude that the behavioural response has a greater contribution to aging than the modified energy consumption in altered gravity environments.     

Effect of magnetically simulated zero-gravity and enhanced gravity on the walk of the common fruitfly

Understanding the effects of gravity on biological organisms is vital to the success of future space missions. Previous studies in Earth orbit have shown that the common fruitfly (Drosophila melanogaster) walks more quickly and more frequently in microgravity, compared with its motion on Earth. However, flight preparation procedures and forces endured on launch made it difficult to implement on the Earth''s surface a control that exposed flies to the same sequence of major physical and environmental changes. To address the uncertainties concerning these behavioural anomalies, we have studied the walking paths of D. melanogaster in a pseudo-weightless environment (0g*) in our Earth-based laboratory. We used a strong magnetic field, produced by a superconducting solenoid, to induce a diamagnetic force on the flies that balanced the force of gravity. Simultaneously, two other groups of flies were exposed to a pseudo-hypergravity environment (2g*) and a normal gravity environment (1g*) within the spatially varying field. The flies had a larger mean speed in 0g* than in 1g*, and smaller in 2g*. The mean square distance travelled by the flies grew more rapidly with time in 0g* than in 1g*, and slower in 2g*. We observed no other clear effects of the magnetic field, up to 16.5 T, on the walks of the flies. We compare the effect of diamagnetically simulated weightlessness with that of weightlessness in an orbiting spacecraft, and identify the cause of the anomalous behaviour as the altered effective gravity.     

The First European Parabolic Flight Campaign with the Airbus A310 ZERO-G

Aircraft parabolic flights repetitively provide up to 23 seconds of reduced gravity during ballistic flight manoeuvres. Parabolic flights are used to conduct short microgravity investigations in Physical and Life Sciences and in Technology, to test instrumentation prior to space flights and to train astronauts before a space mission. The use of parabolic flights is complementary to other microgravity carriers (drop towers, sounding rockets), and preparatory to manned space missions on board the International Space Station and other manned spacecraft, such as Shenzhou and the future Chinese Space Station. After 17 years of using the Airbus A300 ZERO-G, the French company Novespace, a subsidiary of the ’Centre National d’Etudes Spatiales’ (CNES, French Space Agency), based in Bordeaux, France, purchased a new aircraft, an Airbus A310, to perform parabolic flights for microgravity research in Europe. Since April 2015, the European Space Agency (ESA), CNES and the ‘Deutsches Zentrum für Luft- und Raumfahrt e.V.’ (DLR, the German Aerospace Center) use this new aircraft, the Airbus A310 ZERO-G, for research experiments in microgravity. The first campaign was a Cooperative campaign shared by the three agencies, followed by respectively a CNES, an ESA and a DLR campaign. This paper presents the new Airbus A310 ZERO-G and its main characteristics and interfaces for scientific experiments. The experiments conducted during the first European campaign are presented.     

Pipette-based Method to Study Embryoid Body Formation Derived from Mouse and Human Pluripotent Stem Cells Partially Recapitulating Early Embryonic Development Under Simulated Microgravity Conditions

The in vitro differentiation of pluripotent stem cells partially recapitulates early in vivo embryonic development. More recently, embryonic development under the influence of microgravity has become a primary focus of space life sciences. In order to integrate the technique of pluripotent stem cell differentiation with simulated microgravity approaches, the 2-D clinostat compatible pipette-based method was experimentally investigated and adapted for investigating stem cell differentiation processes under simulated microgravity conditions. In order to keep residual accelerations as low as possible during clinorotation, while also guaranteeing enough material for further analysis, stem cells were exposed in 1-mL pipettes with a diameter of 3.5 mm. The differentiation of mouse and human pluripotent stem cells inside the pipettes resulted in the formation of embryoid bodies at normal gravity (1 g) after 24 h and 3 days. Differentiation of the mouse pluripotent stem cells on a 2-D pipette-clinostat for 3 days also resulted in the formation of embryoid bodies. Interestingly, the expression of myosin heavy chain was downregulated when cultivation was continued for an additional 7 days at normal gravity. This paper describes the techniques for culturing and differentiation of pluripotent stem cells and exposure to simulated microgravity during culturing or differentiation on a 2-D pipette clinostat. The implementation of these methodologies along with -omics technologies will contribute to understand the mechanisms regulating how microgravity influences early embryonic development.     

ARADISH - Development of a Standardized Plant Growth Chamber for Experiments in Gravitational Biology Using Ground Based Facilities

Plant development strongly relies on environmental conditions. Growth of plants in Biological Life Support Systems (BLSS), which are a necessity to allow human survival during long-term space exploration missions, poses a particular problem for plant growth, as in addition to the traditional environmental factors, microgravity (or reduced gravity such as on Moon or Mars) and limited gas exchange hamper plant growth. Studying the effects of reduced gravity on plants requires real or simulated microgravity experiments under highly standardized conditions, in order to avoid the influence of other environmental factors. Analysis of a large number of biological replicates, which is necessary for the detection of subtle phenotypical differences, can so far only be achieved in Ground Based Facilities (GBF). Besides different experimental conditions, the usage of a variety of different plant growth chambers was a major factor that led to a lack of reproducibility and comparability in previous studies. We have developed a flexible and customizable plant growth chamber, called ARAbidopsis DISH (ARADISH), which allows plant growth from seed to seedling, being realized in a hydroponic system or on Agar. By developing a special holder, the ARADISH can be used for experiments with Arabidopsis thaliana or a plant with a similar habitus on common GBF hardware, including 2D clinostats and Random Positioning Machines (RPM). The ARADISH growth chamber has a controlled illumination system of red and blue light emitting diodes (LED), which allows the user to apply defined light conditions. As a proof of concept we tested a prototype in a proteomic experiment in which plants were exposed to simulated microgravity or a 90° stimulus. We optimized the design and performed viability tests after several days of growth in the hardware that underline the utility of ARADISH in microgravity research.     

Fish Inner Ear Otolith Growth Under Real Microgravity (Spaceflight) and Clinorotation

Using late larval stages of cichlid fish (Oreochromis mossambicus) we have shown earlier that the biomineralization of otoliths is adjusted towards gravity by means of a neurally guided feedback loop. Centrifuge experiments, e.g., revealed that increased gravity slows down otolith growth. Microgravity thus should yield an opposite effect, i.e., larger than normal otoliths. Consequently, late larval cichlids (stage 14, vestibular system operational) were subjected to real microgravity during the 12 days FOTON-M3 spaceflight mission (OMEGAHAB-hardware). Controls were kept at 1g on ground within an identical hardware. Animals of another batch were subsequently clinorotated within a submersed fast-rotating clinostat with one axis of rotation (2d-clinostat), a device regarded to simulate microgravity. Temperature and light conditions were provided in analogy to the spaceflight experiment. Controls were maintained at 1g within the same aquarium. After all experiments, animals had reached late stage 21 (fish can swim freely). Maintenance under real microgravity during spaceflight resulted in significantly larger than normal otoliths (both lapilli and sagittae, involved in sensing gravity and the hearing process, respectively). This result is fully in line with an earlier spaceflight study in the course of which otoliths from late-staged swordtails Xiphophorus helleri were analyzed. Clinorotation resulted in larger than 1g sagittae. However, no effect on lapilli was obtained. Possibly, an effect was present but too light to be measurable. Overall, spaceflight obviously induces an adaptation of otolith growth, whereas clinorotation does not fully mimic conditions of microgravity regarding late larval cichlids.     

Life History Responses and Feeding Behavior of Microcrustacea in Altered Gravity – Applicability in Bioregenerative Life Support Systems (BLSS)

Manned space missions, as for example to the planet Mars, are a current objective in space exploration. During such long-lasting missions, aquatic bioregenerative life support systems (BLSS) could facilitate independence of resupply from Earth by regenerating the atmosphere, purifying water, producing food and processing waste. In such BLSS, microcrustaceans could, according to their natural role in aquatic ecosystems, link oxygen liberating, autotrophic algae and higher trophic levels, such as fish. However, organisms employed in BLSS will be exposed to high acceleration (hyper- g) during launch of spacecrafts as well as to microgravity (μg) during space travel. It is thus essential that these organisms survive, perform and reproduce under altered gravity conditions. In this study we present the first data in this regard for the microcrustaceas Daphnia magna and Heterocypris incongruens. We found that after hyper-g exposure (centrifugation) approximately one third of the D. magna population died within one week (generally indicating that possible belated effects have to be considered when conducting and interpreting experiments during which hyper-g occurs). However, suchlike and even higher losses could be countervailed by the surviving daphnids’ unaltered high reproductive capacity. Furthermore, we can show that foraging and feeding behavior of D. magna (drop tower) and H. incongruens (parabolic flights) are rarely altered in μg. Our results thus indicate that both species are suitable candidates for BLSS utilized in space.     

Particle conveying under microgravity in a vibrating vessel

This paper presents a numerical study on the conveying of particles in a vibrating vessel under microgravity. Such a vessel is composed of parallel plates with sawtooth wavy surfaces, which are specifically designed to convey particles using simple vibration. The numerical model was validated by good agreement between the simulated and experimental results. Then the effects of key variables, including the vessel geometry, vibration amplitude and frequency and gravity level, were systematically investigated by a series of controlled simulations. The results confirm the optimised design from the previous experiments, and numerically demonstrate that using such a system a steady conveying operation can be achieved under microgravity. The convey rate is positively affected by the vibration amplitude and frequency in a complicated way, which cannot be simply described by the commonly used vibration intensity or velocity amplitude. The gravity level also has a significant effect on the convey rate when it is over 0.001g. The convey rate can be estimated by the product of the average solid fraction and velocity. And the effects of the variables can be better understood through the analyses on these two parameters. Finally, a predictive model is proposed to estimate the convey rate under different operational conditions. The findings are useful for the design of particle conveying techniques for outer space applications.