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.     

The “<Emphasis Type="Italic">Daphnia</Emphasis>” Lynx Mark I Suborbital Flight Experiment: Hardware Qualification at the Drop Tower Bremen

The Drop Tower Bremen, a ground-based facility enabling research under real microgravity conditions, is an excellent platform for testing new types of experimental hardware to ensure full performance when deployed in costly and rare flight opportunities such as suborbital flights. Here we describe the “Daphnia” experiment which will fly on XCOR Aerospace Lynx Mark I and our experience from the hardware tests with the catapult system at the drop tower. The aim of the “Daphnia” experiment is to obtain data on the biological performance of daphnids and predator-prey interactions in microgravity, which are important for the development of aquatic bioregenerative life support systems (BLSS). The experiment consists of two subunits: The first unit is dedicated to predator-prey interactions, where behavioural analysis should reveal if microgravity interfere with prey (Daphnia) detection or feeding and therefore may interrupt the trophic cascade. The functioning of such an artificial food web is indispensable for a long-lasting BLSS suitable for long-duration manned space missions or Earth-based explorations to extreme habitats. The second unit is designed to investigate the impact of microgravity on gene expression and the cytoskeleton in Daphnia. Next to data collection, the real microgravity conditions at the drop tower have helped to identify the weak points of the “Daphnia” experimental hardware and lead to further improvement. Hence, the drop tower is ideal for testing new experimental hardware which is indispensable before the implementation in suborbital flights.     

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.     

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.     

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.     

The Impact of Hypergravity and Vibration on Gene and Protein Expression of Thyroid Cells

Experiments in space either on orbital missions on-board the ISS, or in suborbital missions using sounding rockets, like TEXUS as well as parabolic flight campaigns are still the gold standard to achieve real microgravity conditions in the field of gravitational biology and medicine. However, during launch, and in flight, hypergravity and vibrations occur which might interfere with the effects of microgravity. It is therefore important to know these effects and discriminate them from the microgravity effects. This can be achieved by ground-based facilities like centrifuges or vibration platforms. Recently, we have conducted several experiments with different thyroid cancer cell lines. This study, as part of the ESA-CORA-GBF 2010-203 project, focused on the influence of vibration and hypergravity on benign human thyroid follicular epithelial cells (Nthy-ori 3-1 cell line). Gene and in part protein expression regulation under both conditions were analyzed for VCAN, ITGA10, ITGB1, OPN, ADAM19, ANXA1, TNFA, ABL2, ACTB, PFN2, TLN1, EZR, RDX, MSN, CTGF, PRKCA, and PRKAA1 using quantitative real-time PCR and Western Blot. We found that hypergravity and vibration affected genes and proteins involved in the extracellular matrix, the cytoskeleton, apoptosis, cell growth and signaling. Vibration always led to a down-regulation, whereas hypergravity resulted in a more heterogeneous expression pattern. Overall we conclude that both conditions can influence gene regulation and production of various genes and proteins. As a consequence, it is important to perform control experiments on hypergravity and vibration facilities in parallel to flight experiments.     

Comprehensive Study of the Influence of Altered Gravity on the Oxidative Burst of Mussel (<Emphasis Type="Italic">Mytilus edulis</Emphasis>) Hemocytes

Microgravity induces alterations in the functioning of immune cell; however, the underlying mechanisms have not yet been identified. In this study, hemocytes (blood cells) of the blue mussel Mytilus edulis were investigated under altered gravity conditions. The study was conducted on the ground in preparation for the BIOLAB TripleLux-B experiment, which will be performed on the International Space Station (ISS). On-line kinetic measurements of reactive oxygen species (ROS) production during the oxidative burst and thus cellular activity of isolated hemocytes were performed in a photomultiplier (PMT)-clinostat (simulated microgravity) and in the 1g operation mode of the clinostat in hypergravity on the Short-Arm Human Centrifuge (SAHC) as well as during parabolic flights. In addition to studies with isolated hemocytes, the effect of altered gravity conditions on whole animals was investigated. For this purpose, whole mussels were exposed to hypergravity (1.8 g) on a multi-sample incubator centrifuge (MuSIC) or to simulated microgravity in a submersed clinostat. After exposure for 48 h, hemocytes were taken from the mussels and ROS production was measured under 1 g conditions. The results from the parabolic flights and clinostat studies indicate that mussel hemocytes respond to altered gravity in a fast and reversible manner. Hemocytes (after cryo-conservation) exposed to simulated microgravity (μ g), as well as fresh hemocytes from clinorotated animals, showed a decrease in ROS production. Measurements during a permanent exposure of hemocytes to hypergravity (SAHC) show a decrease in ROS production. Hemocytes of mussels measured after the centrifugation of whole mussels did not show an influence to the ROS response at all. Hypergravity during parabolic flights led to a decrease but also to an increase in ROS production in isolated hemocytes, whereas the centrifugation of whole mussels did not influence the ROS response at all. This study is a good example how ground-based facility experiments can be used to prepare for an upcoming ISS experiment, in this case the TRIPLE LUX B experiment.     

Synthesis and study of Np(V) and Pu(V) benzoate complexes with bipyridine

Heteroligand compounds AnO₂(bipy)OOCC₆H₅ (An = Np, Pu; bipy = α,α-bipyridine, C¹⁰H⁸N²) were synthesized and studied. It follows from powder X-ray patterns that these compounds are isostructural. Their unit cell parameters, determined by indexing of the powder X-ray patterns, are as follows: a = 9.2162 (7), b = 10.2339(8), c = 17.4083(17) Å, and β = 96.48(1)° for Np and a = 9.1983(18), b = 10.2052(18), c = 17.370(3) Å and β = 96.51(1)° for Pu. The compounds crystallize in the monoclinic system space group P2₁/n, Z = 4. The electronic absorption spectra of crystalline compounds suggest pentagonal-bipyramidal surrounding of the central atom and the prescence of cation-cation bonds with AnO ₂ ⁺ ions acting as monodentate ligands with respect to each other. The IR spectra of the compounds were recorded, and their thermal behavior in air was studied.     

Simulating Parabolic Flight like <Emphasis Type="Italic">g</Emphasis>-Profiles on Ground - A Combination of Centrifuge and Clinostat

Clinostats and centrifuges are widely used to create simulated microgravity or hypergravity, respectively, in order to study the impact of gravity on biosystems. Here, we used a clinostat and a centrifuge in alternating modes of operation in order to create a simulated parabolic flight like g-profile. To our knowledge, it is the first time that both devices were run in connection. In order to test the method, we investigated the production of reactive oxygen species of immune cells (macrophages) during oxidative burst in an on-line kinetic approach, which has been extensively studied under real (parabolic flight) and simulated microgravity (clinostat) as well as under hypergravity conditions (centrifuge). Our results indicate that clinostat and centrifuge can be operated in an alternating way to simulate the repetitive changes of gravity during parabolic flight. Although the switch from one gravity level to the other could not be carried out as quickly as it takes place during actual parabolic flight due to technical and operational reasons, it can be concluded that running experiments in a clinostat aboard a centrifuge on ground are suitable for studying gravity-related phenomena.     

Annotated Gene and Proteome Data Support Recognition of Interconnections Between the Results of Different Experiments in Space Research

In a series of studies, human thyroid and endothelial cells exposed to real or simulated microgravity were analyzed in terms of changes in gene expression patterns or protein content. Due to the limitation of available cells in many space research experiments, comparative and control experiments had to be done in a serial manner. Therefore, detected genes or proteins were annotated with gene names and SwissProt numbers, in order to allow searches for interconnections between results obtained in different experiments by different methods. A crosscheck of several studies on the behavior of cytoskeletal genes and proteins suggested that clusters of cytoskeletal components change differently under the influence of microgravity and/or vibration in different cell types. The result that LOX and ISG15 gene expression were clearly altered during the Shenzhou-8 spaceflight mission could be estimated by comparison with the results of other experiments. The more than 100-fold down-regulation of LOX supports our hypothesis that the amount and stability of extracellular matrix have a great influence on the formation of three-dimensional aggregates under microgravity. The approximately 40-fold up-regulation of ISG15 cannot yet be explained in detail, but strongly suggests that ISGylation, an alternative form of posttranslational modification, plays a role in longterm cultures.     

X-ray analysis of 2-aniline benzo(2,3-<Emphasis Type="Italic">b</Emphasis>) cyclopentane-1,3-dione

The molecular and crystal structure of 2-aniline benzo(2,3-b) cyclopentane-1,3-dione has been determined by X-ray crystallographic techniques. This compound crystallizes in the orthorhombic space groupP2₁2₁2₁ with unit cell parameters:a = 5.467(1),b = 10.657(3),c = 19.602(6) Å;V = 1142.01(5) ų,Z = 4. The crystal structure has been resolved up to anR-factor 0.050 for 1129 reflections. All the three rings in the structure are planar. However, the dihedral angle between the phenyl ring and the moiety comprising of a five-membered and six-membered ring is 92.4°. The oxygen atom O1 acts as a trifurcated acceptor and is involved in the formation of three intermolecular interactions.     

Laser Interferometer for Space Gravitational Waves Detection and Earth Gravity Mapping

The idea of using space laser interferometer to measure the relative displacement change between two satellites has been considered for space gravitational waves detection and Earth gravity filed mapping in recent years. Some investigations on the key issues of laser interferometer in our working team have been presented in this paper. An on-ground laser interferometer prototype used for the demonstration of satellite-to-satellite ranging has been constructed, which is equipped with phasemeter, laser pointing modulation and laser phase-locking control. The experimental results show that path-length measurement sensitivity of the laser interferometer reaches 200 pm/√ Hz, and phase measurement precision achieves 2π × 10^??5 rad/√ Hz, and laser pointing modulation precision is better than 80 nrad/√ Hz, and laser phase-locking control precision attains 2π × 10^??4 rad/√ Hz within the frequency regime of 1 mHz–1 Hz. All of these demonstrate that the proposed laser interferometer has very promising feasibility to meet the requirement of the Taiji, TianQin and Space Advanced Gravity Measurement (SAGM) missions which are put forward by Chinese scientists.     

Experimental Study of the Thermodynamic Properties of Diethyl Ether (DEE) at Saturation

The isochoric heat capacities \({({C_{V1}^{\prime}} ,{C_{V1}^{\prime\prime}},{C_{V2}^{\prime}},{C_{V2}^{\prime\prime}})}\), saturation densities (\({\rho _{\rm S}^{\prime}}\) and \(({\rho_{\rm S}^{\prime\prime})}\)), vapor pressures (P _S), thermal-pressure coefficients \({\gamma_V=\left({\partial P/\partial T}\right)_V}\), and first temperature derivatives of the vapor pressure γ _S = (dP _S/dT) of diethyl ether (DEE) on the liquid–gas coexistence curve near the critical point have been measured with a high-temperature and high-pressure nearly constant-volume adiabatic piezo-calorimeter. The measurements of \({({C_{V1}^{\prime}} ,{C_{V1}^{\prime\prime}},{C_{V2}^{\prime}},{C_{V2}^{\prime\prime}})}\) were made in the liquid and vapor one- and two-phase regions along the coexistence curve. The calorimeter was additionally supplied with a calibrated extensometer to accurately and simultaneously measure the PVT, C _V VT, and thermal-pressure coefficient, γ _V , along the saturation curve. The measurements were carried out in the temperature range from 416 K to 466.845 K (the critical temperature) for 17 liquid and vapor densities from 212.6 kg · m^?3 to 534.6 kg · m^?3. The quasi-static thermo- (reading of PRT, T ? τ plot) and baro-gram (readings of the tensotransducer, P ? τ plot) techniques were used to accurately measure the phase-transition parameters (P _S ,ρ _S ,T _S) and γ _V . The total experimental uncertainty of density (ρ _S), pressure (P _S), temperature (T _S), isochoric heat capacities \({({C_{V1}^{\prime}} ,{C_{V1}^{\prime\prime}},{C_{V2}^{\prime}},{C_{V2}^{\prime\prime}})}\), and thermal-pressure coefficient, γ _V , were estimated to be 0.02 % to 0.05 %, 0.05 %, 15 mK, 2 % to 3 %, and 0.12 % to 1.5 %, respectively. The measured values of saturated caloric \({({C_{V1}^{\prime}} ,{C_{V1}^{\prime\prime}},{C_{V2}^{\prime}},{C_{V2}^{\prime\prime}})}\) and saturated thermal (P _S, ρ _S, T _S) properties were used to calculate other derived thermodynamic properties C _P , C _S, W, K _T , P _int, ΔH _vap, and \({\left({\partial V/\partial T}\right)_P^{\prime}}\) of DEE near the critical point. The second temperature derivatives of the vapor pressure, (d² P _S/dT ²), and chemical potential, (d² μ/dT ²), were also calculated directly from the measured one- and two-phase liquid and vapor isochoric heat capacities \({({C_{V1}^{\prime}} ,{C_{V1}^{\prime\prime}},{C_{V2}^{\prime}},{C_{V2}^{\prime\prime}})}\) near the critical point. The derived values of (d² P _S/dT ²) from calorimetric measurements were compared with values calculated from vapor–pressure equations. The measured and derived thermodynamic properties of DEE near the critical point were interpreted in terms of the “complete scaling” theory of critical phenomena. In particular, the effect of a Yang–Yang anomaly of strength R _μ on the coexistence-curve diameter behavior near the critical point was studied. Extended scaling-type equations for the measured properties P _S (T), ρ _S (T), and \({({C_{V1}^{\prime}} ,{C_{V1}^{\prime\prime}},{C_{V2}^{\prime}},{C_{V2}^{\prime\prime}})}\) as a function of temperature were developed.     

First-Principle Study of Electronic and Half-Metallic Ferromagnetic Properties of Vanadium (V)-Doped Cubic BP and InP

In this study, we use the first-principle calculations of density functional theory with gradient generalized approximation of Wu–Cohen to investigate the doping effect of vanadium impurity on structural, electronic and magnetic properties of In_1?x V _x P and B_1?x V _x P alloys at various concentrations x = 0.0625, 0.125 and 0.25. Owing to the metallic nature of majority spin and semiconducting minority spin, the In_1?x V _x P compounds exhibit a half-metallic character with total magnetic moments of 2 μ _B, while the B_1?x V _x P has metallic nature for all concentrations. The results of exchange parameters revealed that exchange coupling between vanadium atoms and the conduction band is ferromagnetic, confirming the magnetic feature of In_1?x V _x P and B_1?x V _x P. From our findings, we have predicted that the In_1?x V _x P alloys seem to be potential materials for spintronics.     

4<Emphasis Type="Italic">H</Emphasis>-SiC Based Subnanosecond (150 ps) High-Voltage (1600 V) Current Breakers

High-voltage (1600 V) diodes based on epitaxial 4H-SiC p⁺⁺–p⁺–n₀–n⁺ structures are tested as fast current breakers included in a special pulsed circuit. The measured current-breakdown time is about 150 ps. This is a record short time for high-voltage (above 1000 V) silicon-carbide diode breakers. The saturated hole-drift velocity in 4H-SiC of p type is experimentally estimated for the first time: v _sp = 3 × 10⁶ cm/s.     

Novel Solid-State Growth of <Emphasis Type="Italic">p</Emphasis>-Terphenyl: the Parent High-<Emphasis Type="Italic">T</Emphasis><Subscript>c</Subscript> Organic Superconductor (HTOS)

We report an easy and versatile route for the synthesis of the parent phase of the newest superconducting wonder material, i.e., p-terphenyl. Doped p-terphenyl has recently shown superconductivity with transition temperature as high as 120 K. For crystal growth, the commercially available p-terphenyl powder is pelletized, encapsulated in an evacuated (10^?4 Torr) quartz tube and subjected to high-temperature (260 ^°C) melt followed by slow cooling at 5 ^°C/h. A simple temperature-controlled heating furnace is used during the process. The obtained crystal is one piece, shiny, and plate like. Single crystal surface XRD (X-ray diffraction) showed unidirectional (00l) lines, indicating that the crystal is grown along the c-direction. Powder XRD of the specimen showed that as grown p-terphenyl is crystallized in monoclinic structure with space group P2 ₁/a space group, having lattice parameters a = 7.672 (2) Å, b = 5.772 (5) Å, and c = 13.526(3) Å and β = 91.484 (3)^°. Scanning electron microscopy (SEM) pictures of the crystal showed clear layered slab-like growth without any visible contamination from oxygen. Characteristic reported Raman active modes related to C–C–C bending, C–H bending, C–C stretching, and C–H stretching vibrations are seen clearly for the studied p-terphenyl crystal. The physical properties of the crystal are yet underway. The short letter reports an easy and versatile crystal growth method for obtaining quality p-terphenyl. The same growth method may probably be applied to doped p-terphenyl and to subsequently achieve superconductivity to the tune of as high 120 K for the newest superconductivity wonder, i.e., high- T _c organic superconductor (HTOS).