Modeled microgravity increases filamentation, biofilm formation, phenotypic switching, and antimicrobial resistance in Candida albicans

Candida albicans is an opportunistic fungal pathogen responsible for a variety of cutaneous and systemic human infections. Virulence of C. albicans increases upon exposure to some environmental stresses; therefore, we explored phenotypic responses of C. albicans following exposure to the environmental stress of low-shear modeled microgravity. Upon long-term (12-day) exposure to low-shear modeled microgravity, C. albicans transitioned from yeast to filamentous forms at a higher rate than observed under control conditions. Consistently, genes associated with cellular morphology were differentially expressed in a time-dependent manner. Biofilm communities, credited with enhanced resistance to environmental stress, formed in the modeled microgravity bioreactor and had a more complex structure than those formed in control conditions. In addition, cells exposed to low-shear modeled microgravity displayed phenotypic switching, observed as a near complete transition from smooth to "hyper" irregular wrinkle colony morphology. Consistent with the presence of biofilm communities and increased rates of phenotypic switching, cells exposed to modeled microgravity were significantly more resistant to the antifungal agent Amphotericin B. Together, these data indicate that C. albicans adapts to the environmental stress of low-shear modeled microgravity by demonstrating virulence-associated phenotypes.     

Altered expression of mRNAs implicated in osteogenesis under conditions of simulated microgravity is regulated by CD200:CD200R

Mouse calvarial cells grown under simulated microgravity conditions (neutral buoyancy) show preferential differentiation towards the osteoclast lineage, as defined by surrogate mRNAs, bone nodule growth and TRAP⁺ cells, when compared with cells cultured under normal gravity conditions. This effect was suppressed in cultures which contained the immunoregulatory molecule CD200, and conversely enhanced by anti-CD200 mAb. Concomitant increases occur in expression of inflammatory cytokines, and their mRNAs, under simulated microgravity conditions. Again cultures containing exogenous CD200 showed suppressed cytokine and cytokine mRNA expression. Further alterations in osteoclastogenesis were seen using cells isolated from cytokine-receptor knockout mice. We conclude that, as assessed by altered expression of mRNAs associated with osteoblast differentiation, CD200:CD200R interactions play an important regulatory role in the enhanced osteoclastogenesis seen under simulated microgravity conditions, with changes in cytokine expression further modulating this effect.     

Separation of bacterial cells by free flow electrophoresis under microgravity: a result of the SpaceLab-Japan project on Space Shuttle flight STS-47

We demonstrated free flow electrophoresis (FFE) of charged cells under microgravity, where gravitational effects are almost eliminated. Separation of a mixture of three bacterial strains (mutants of Salmonella typhimurium LT2) by FFE was conducted on NASA Space Shuttle flight STS-47 (September 1992). The experiment was designed to differentiate three strains having different lipopolysaccharide core structures in the cell membrane. The results were compared to those of ground experiments, in order to examine whether or not FFE in a weightless environment provides distinct advantages. Smooth strain SL1027 and rough strain SL3749 migrated to two separated fractions. The quality (viability) and the yields of the separated samples were sufficient to show the advantage of microgravity. Another rough strain, SL1102, exhibited unexpected electrophoretic behavior, which prevented the complete resolution of the three strains. All the strains were recovered as viable cells after 8 days of flight. The present study suggests that electrophoretic separation of bacterial cells is much more effective under microgravity conditions with relatively good resolution in comparison with the ground operation.     

Fluid flow interaction with an obstacle near free surface

The two-dimensional problem of thin obstacle interaction with compressible fluid flow near free surface under microgravity was regarded. The fluid was assumed occupying infinite semi-space, gravity was neglected as compared with fluid inertia. The obtained exact solution shows that for relatively thin layer the resistance force does not depend on the layer thickness, but depend on body length and inclination angle, and surpasses twice the respective values for the infinite medium, which could be explained by additional wave resistance arising in the presence of free surface.     

Gravitational Sensitivity of Melts at the Growth of InSb:Te Crystals by the Bridgman and Floating Zone Methods under the Conditions of Microgravity

The comparative analysis of the results of space and ground-based experiments IMET RAS on the growth of InSb:Te crystals by the Bridgman method and floating zone method (FZM) is made for the purpose of studying the influence of microgravity on the growth, structure, and properties of grown crystals, and thus the gravity sensitivity of InSb melt is demonstrated. It is shown that, under microgravity conditions, the Bridgman method makes it possible to grow InSb:Te crystals without contact with the ampoule walls, which provides for the single crystal structure, the absence of striations, and a low dislocation density. For the first time, InSb:Te monocrystals were grown with the FZM under microgravity. The anomalous behavior of the impurity core (facet effect) in these crystals correlates with the changed magnitude and direction of the quasi-stationary (residual) microaccelerations.     

Taxane recovery from cells of Taxus in micro- and hypergravity

Cell suspension cultures of Taxus cuspidata produce taxanes that are released from the outer surface of cells into the culture medium as free and bound alkaloids. Paclitaxel (Taxol (TM)), is an anti-cancer drug in short supply. It has a taxane ring derived from baccatin III and a C-13 phenylisoserine side-chain. This drug is produced over a wide range of gravitational forces. Monoclonal and polyclonal antibodies to paclitaxel, baccatin III, and the C-13 phenylisoserine side chain were combined in multiple-labeling studies to localize taxanes and paclitaxel on cell surfaces or on particles released into the culture medium. Bioreactor vessel design altered the composition of taxanes recovered from cells in simulated microgravity. At 10(-2) and 2x10(-4)g, taxane recovery was reduced but biomass growth and percent paclitaxel was significantly increased. At 1 to 24g, growth was reduced with a significant recovery of total taxanes with low percent paclitaxel. Bound paclitaxel was also localized in endonuclease-rich fragmenting nuclei of individual apoptotic cells. A model is presented comprising TCH (touch) genes encoding enzymes that modify taxane-bearing xylan residues in cell walls, the calcium-sensing of gravitational forces by the cytoplasm, and the predisposition of nuclei to apoptosis. This integrates the adaptive physiological and biochemical responses of drug-producing genomes with gravitational forces.     

Adaptive environmental control for optimal results during plant microgravity experiments

The SVET Space Greenhouse (SG)--the first and the only automated plant growth facility onboard the MIR Space Station in the period 1990-2000 was developed on a Russian-Bulgarian Project in the 80s. The aim was to study plant growth under microgravity in order to include plants as a link of future Biological Life Support Systems for the long-term manned space missions. An American developed Gas Exchange Measurement System (GEMS) was added to the existing SVET SG equipment in 1995 to monitor more environmental and physiological parameters. A lot of long-duration plant flight experiments were carried out in the SVET+GEMS. They led to significant results in the Fundamental Gravitational Biology field--second-generation wheat seeds were produced in the conditions of microgravity. The new International Space Station (ISS) will provide a perfect opportunity for conducting full life cycle plant experiments in microgravity, including measurement of more vital plant parameters, during the next 15-20 years. Nowadays plant growth facilities for scientific research based on the SVET SG functional principles are developed for the ISS by different countries (Russia, USA, Italy, Japan, etc.). A new Concept for an advanced SVET-3 Space Greenhouse for the ISS, based on the Bulgarian experience and "know-how" is described. The absolute and differential plant chamber air parameters and some plant physiological parameters are measured and processed in real time. Using the transpiration and photosynthesis measurement data the Control Unit evaluates the plant status and performs adaptive environmental control in order to provide the most favorable conditions for plant growth at every stage of plant development in experiments. A conceptual block-diagram of the SVET-3 SG is presented.     

Floating zone experiments with germanium crystals in sounding rockets

A floating zone experiment with a coated surface was performed in a sounding rocket. Analysis of the segregation showed that elimination of free surfaces decreased the convection in the liquid zone. Analysis of radial segregation during floating zone crystal growth under different experimental conditions demonstrated high sensitivity to interface shape and to convection. Under conditions of very weak convection, microgravity and coated surfaces, dopant concentration fields giving severe radial segregation were built up.     

The properties of alamethicin incorporated into planar lipid bilayers under the influence of microgravity.

During evolution, life on earth had adapted to the gravity of 1g. Due to space flight, in the last decades the question arose what happens to the brain under microgravity on the molecular level. Ion channels among others are the molecular basis of brain function. Therefore, the investigation of ion channel function under microgravity seems to be a promising approach to gather knowledge on brain function during space flight. In a first step, the ion channel forming peptide Alamethicin was used as a model channel in an artificial membrane. It is well suitable for this kind of investigation, since its properties are well described under standard gravity. For that reason, changes due to microgravity can be detected easily. All experiments were performed in the German drop tower at ZARM-FAB, Bremen. A special set-up was constructed based on the bilayer technique introduced by Mueller and Rudin. All functions of this set-up can be observed and controlled remotely. In the first set of experiments, a dramatic change of electrical properties of Alamethicin under microgravity could be observed. Mainly, the pore frequency is significantly reduced.     

Evaluation of bleomycin-induced chromosome aberrations under simulated microgravity conditions in human lymphocytes using "FISH" techniques

In the present investigation we report the effects of simulated microgravity conditions (clinostat) on the induction of chromosomal aberrations in human lymphocytes in vitro by (R) Bleomycin. Chromosomal aberrations have been analysed by means of fluorescent in situ hybridisation (FISH) and chromosome-specific composite DNA probes (chromosome painting). The results obtained show that, under simulated microgravity conditions, the levels of both symmetrical and asymmetrical (dicentrics, rings), the number of cells bearing "complex" aberrations and hence the total numbers of aberrations were significantly elevated at any of the dose-levels assayed, compared to the parallel treatments performed as 1g control ("ground"). Furthermore, the ratio symmetrical:asymmetrical translocations was markedly elevated under simulated microgravity conditions, compared to the findings usually observed under "normal" 1g conditions. On these bases, we are much inclined to believe that simulated microgravity, rather than limiting the resealing of DNA double strand breaks (DSB's) induced by genotoxic agents is influencing in terms of enhancement the misrejoining of DSB's which is actually responsible for the fixation of the original lesions to DNA into chromosomal aberrations. In addition, the possible different misrepair processes leading to the formation of symmetrical and asymmetrical translocations might be differentially influenced by microgravity being the symmetrical translocations significantly more represented.     

The effects of low-shear mechanical stress on Yersinia pestis virulence

Manned space exploration has created a need to evaluate the effects of spacelike stress on pathogenic and opportunistic microbes astronauts could carry with them to the International Space Station and beyond. Yersinia pestis (YP) causes bubonic, septicemic, and pneumonic plague and is capable of killing infected patients within 3-7 days. In this study, low-shear modeled microgravity (LSMMG), a spacelike stress, was used to physically stress YP; and its effects on proliferation, cold growth, and type III secretion system (T3SS) function were evaluated. YP was grown to saturation in either LSMMG or normal gravity (NG) conditions prior to being used for RAW 246.7 cell infections, HeLa cell infections, and Yop secretion assays. A mutant strain of YP (ΔyopB) that lacks the ability to inject Yersinia outer membrane proteins (Yops) into the host cell was used as a negative control in cell infection experiments. Our experimental results indicate that YP cultivated under LSMMG resulted in reduced YopM production and secretion compared to its NG-grown counterpart. Similarly, NG-grown YP induced more cell rounding in HeLa cells than did the LSMMG-grown YP, which suggests that LSMMG somehow impairs T3SS optimum function. Also, LSMMG-grown YP used to infect cultured RAW 246.7 cells showed a similar pattern of dysfunction in that it proliferated less than did its NG-grown counterpart during an 8-hour infection period. This study suggests that LSMMG can attenuate bacterial virulence contrary to previously published data that have demonstrated LSMMG-induced hypervirulence of other Gram-negative enterics.     

Body orientation and center of mass control in microgravity

The control of the body orientation and the center of mass position with respect to the feet was investigated under normo- and microgravity (space flight Altair), during erect posture and at the end of a forward or backward upper trunk movement.

It was observed that during erect posture, the trunk orientation with respect to the vertical was inclined some 6 ° forward in both subjects under microgravity, whereas it was vertical or slightly backward oriented under normogravity. Under microgravity, on the contrary, the initial position CM changed either backwards or forwards. This result suggests that the inclined trunk posture might be due to misevaluating the vertically under microgravity and that different control mechanisms are involved in orienting the trunk and placing the CM.

It was also noted that the final position of the CM at the end of the movement did not differ markedly between microgravity and normogravity. This result suggests that the kinematic synergies which stabilize the CM during uppertrunk movements may result from an automatic central control which is independent from the gravity constraints.     

An endogenous growth pattern of roots is revealed in seedlings grown in microgravity

In plants, sensitive and selective mechanisms have evolved to perceive and respond to light and gravity. We investigated the effects of microgravity on the growth and development of Arabidopsis thaliana (ecotype Landsberg) in a spaceflight experiment. These studies were performed with the Biological Research in Canisters (BRIC) hardware system in the middeck region of the space shuttle during mission STS-131 in April 2010. Seedlings were grown on nutrient agar in Petri dishes in BRIC hardware under dark conditions and then fixed in flight with paraformaldehyde, glutaraldehyde, or RNAlater. Although the long-term objective was to study the role of the actin cytoskeleton in gravity perception, in this article we focus on the analysis of morphology of seedlings that developed in microgravity. While previous spaceflight studies noted deleterious morphological effects due to the accumulation of ethylene gas, no such effects were observed in seedlings grown with the BRIC system. Seed germination was 89% in the spaceflight experiment and 91% in the ground control, and seedlings grew equally well in both conditions. However, roots of space-grown seedlings exhibited a significant difference (compared to the ground controls) in overall growth patterns in that they skewed to one direction. In addition, a greater number of adventitious roots formed from the axis of the hypocotyls in the flight-grown plants. Our hypothesis is that an endogenous response in plants causes the roots to skew and that this default growth response is largely masked by the normal 1?g conditions on Earth.     

表面张力贮箱的微重力实验验证--静平衡与重定位

新研制的表面张力贮箱保证在微重力或无重力环境下、在任何时候都能向系统提供不含气的推进剂，因而需要确定液体的空间分布以及在给定微重力加速度下的重定位过程，以便为贮箱的设计提供依据。为此，按照相似理论用缩比模型进行了一系列微重力落塔实验，然后根据测定的重定位时间计算出原型表面张力贮箱的重定位时间。     

Effects of buoyancy on gas-jet diffusion flames: Experiment and theory

Theoretical and experimental research on the effects of buoyancy on gas-jet diffusion flames is described in this paper. Part of this research involves an assessment of existing data obtained under reduced-gravity conditions. The results show that uncertainties in our understanding of flame structure exist and further research is required before reliable predictions of ignition, stabilization and propagation of flames under microgravity conditions can be made. Steady-state and transient theories have been developed and used in the analysis of existing drop-tower data and new data obtained from a stationary experiment involving inverted flames. The result of this research has led to the definition of a microgravity experiment to be performed in space.     

The Protein Crystallization Facility (PCF) for EURECA.

Research on the structure of molecules by X-ray diffraction analysis requires large single crystals. However, the dynamic behavior of proteins caused by their high molecular weight prevents the growth of large single crystals if this process is disturbed by thermal convection. For example, protein single crystals grown under terrestrial (1 g) conditions are limited to dimensions in the order of 0.1 mm, whereas the size of crystals, grown under (quasi) space conditions has been 5 times larger (pilot experiment CRYOSTAT, Spacelab). Under EURECA conditions (e.g. no microgravity disturbances), the result in regularity of crystal growth and size is expected to be much better. In this paper an overview is given of the protein crystallization facility which includes Experiment-, Service- and Secondary Cooling Module, and its interfaces to the EURECA Carrier. At the end, there will be presented a short mission profile concerning cooling-, power- and data exchange requirements.     

微重力下热薄材料燃烧特性的窄通道实验研究

利用高度分别为10 mm,12 mm和14 mm的水平窄通道对微重力环境下热薄材料表面的火焰传     

Experimental Study of the Structures of Macroparticles in the Dust Plasma under Microgravity Conditions onboard the MirOrbital Complex

The dynamics of formation of the ordered structures of charged macroparticles under microgravity conditions is investigated. The experimental observations of the behavior of an ensemble of macroparticles were carried out onboard the Mirspace station. The analysis and comparison of results of experimental and theoretical investigations allow us to conclude that under microgravity conditions the formation of elongated, ordered structures of macroparticles, charged by solar radiation, is possible.     

Effects of Local Detachment of a Melt from Ampoule Walls,Crystal Vibration,and Magnetic Field at Crystal Growth under Zero Gravity

A numerical investigation of the melt flow and heat and mass transfer is carried out at the crystal growth under zero gravity, when the melt detachment from ampoule walls, crystal vibration, and various magnetic fields are active. Specific features of the melt flow are demonstrated depending on the size of a detachment zone adjacent to the crystallization boundary. The velocity of the averaged flow generated by crystal vibration is determined as a function of the vibration intensity. It is shown that the crystal vibration cannot compensate a thermal capillary flow (caused by detachment of the melt from the ampoule wall) and reduce the macrosegregation of impurities. It is shown that the application of steady and rotating magnetic fields are inefficient for all ampoule methods of crystal growth under microgravity conditions.