Clinorotation Increases the Growth of Utricular Otoliths of Developing Cichlid Fish

It has been shown earlier that hypergravity slows down inner ear otolith growth in developing fish as an adaptation towards increased environmental gravity. Suggesting that otolith growth is regulated by the central nervous system, thus adjusting otolithic weight to produce a test mass, applying functional weightlessness should yield an opposite effect, i.e. larger than normal otoliths. Therefore, larval siblings of cichlid fish (Oreochromis mossambicus) were housed for 7 days in a submersed, two-dimensional clinostat, which provided a residual gravity of approximately 0.007g. After the experiment, otoliths were dissected and their size (area grown during the experiment) was determined. Maintenance in the clinostat resulted in significantly larger utricular otoliths (lapilli, involved in graviperception). There were no statistical significant differences regarding saccular otoliths obtained (sagittae, involved in transmitting linear acceleration and, especially, in the hearing process). These results indicated, that the animals had in fact received functional weightlessness. In line and contrasting results on the otoliths of other teleost species kept at actual microgravity (spaceflight) or within rotating wall vessels are discussed.     

Hypergravity of 10<Emphasis Type="Italic">g</Emphasis> Changes Plant Growth,Anatomy, Chloroplast Size,and Photosynthesis in the Moss <Emphasis Type="Italic">Physcomitrella patens</Emphasis>

The photosynthetic and anatomical responses of bryophytes to changes in gravity will provide crucial information for estimating how these plant traits evolved to adapt to changes in gravity in land plant history. We performed long-term hypergravity experiments at 10g for 4 and 8 weeks using the moss Physcomitrella patens with two centrifuges equipped with lighting systems that enable long-term plant growth under hypergravity with irradiance. The aims of this study are (1) to quantify changes in the anatomy and morphology of P. patens, and (2) to analyze the post-effects of hypergravity on photosynthesis by P. patens in relation to these changes. We measured photosynthesis by P. patens for a population of gametophores (e.g., canopy) in Petri dishes and plant culture boxes. Gametophore numbers increased by 9% for a canopy of P. patens, with 24–27% increases in chloroplast sizes (diameter and thickness) in leaf cells. In a canopy of P. patens, the area-based photosynthesis rate (A _canopy) was increased by 57% at 10g. The increase observed in A _canopy was associated with greater plant numbers and chloroplast sizes, both of which involved enhanced CO₂ diffusion from the atmosphere to chloroplasts in the canopies of P. patens. These results suggest that changes in gravity are important environmental stimuli to induce changes in plant growth and photosynthesis by P. patens, in which an alteration in chloroplast size is one of the key traits. We are now planning an ISS experiment to investigate the responses of P. patens to microgravity.     

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 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.     

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.     

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.     

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.     

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.     

Effects of Short-Term Hypergravity Exposure are Reversible in <Emphasis Type="Italic">Triticum aestivum</Emphasis> L. Caryopses

Short-term hypergravity exposure is shown to retard seed germination, growth and photosynthesis in wheat caryopses. This study investigates the reversibility of effects of short-term hypergravity on imbibed wheat (Triticum aestivum var L.) caryopses. After hypergravity exposure (500 × g ? 2500 × g for 10 min) on a centrifuge, exposed caryopses were kept under normal gravity (1 × g) up to six days and then sown on agar. Results of the present study showed that percentage germination and growth were completely restored for DAY 6 compared to DAY 0. Restoration of germination and growth was accompanied by increased α-amylase activity. The specific activity of antioxidative enzyme viz. catalase and guaiacol peroxidase was lowered on DAY 6 compared to DAY 0 suggesting an alleviation of oxidative cellular damage against hypergravity stress. Chlorophyll pigment recovery along with chlorophyll fluorescence (PI and Fv/Fm) on DAY 6 indicates a transient rather than permanent damage of the photosynthetic apparatus. Thus, our findings demonstrate that short-term hypergravity effects are reversible in wheat caryopses. The metabolic cause of restoration of seed germination and growth upon transferring the caryopses to normal gravity is performed by a reactivation of carbohydrate- metabolizing enzymes, α-amylase and alleviation of oxidative stress damage with subsequent recovery of chlorophyll biosynthesis and photosynthetic activity.     

Effect of γ-irradiation on the frequency-independent parameters of an equivalent circuit for two-dimensional island platinum films

Four sets (A, B, C and D) of two-dimensional island platinum films [2D-I(Pt)Fs] were prepared by the thermal evaporation technique. The mass thicknesses for the films of such sets are 1, 5, 10 and 20 Å, respectively. The frequency-independent parameters of one of the equivalent circuit for island metal films in general are R _b, R _g and C _g where R _b is the resistance of the islands themselves while R _g is the resistance that accompanies the transfer of electrons across the gaps among islands and C _g is the capacitance that results from the separated metallic islands. The least-square values of R _b, R _g and C _g for our films were deduced via a computerized analysis. The stabilized films were exposed to γ-rays at doses 100, 200, 300, 500 and 700 Gy using ¹³⁷Cs (0.662 MeV) radiation source of dose rate 0.5 Gy/min. It was found that (1) For any particular mass thickness: (a) R _b and R _g decreases with the increase in the dose of γ-rays while C _g increases (b) the absolute value of the fractional change in R _b, R _g and C _g increases with the increase in dose. (2) For any particular dose the increase in the fractional change in R _b, R _g and C _g becomes more pronounced when the mass thickness increases. Qualitative interpretation for our results was given on the ground that γ-rays changed the island’s shape from spherical to prolate spheroid one and this has been checked experimentally via imaging the films by Atomic Force Microscope before and after γ-irradiations.     

Development of new ionic gelation strategy: Towards the preparation of new monodisperse and stable hyaluronic acid/β-cyclodextrin-grafted chitosan nanoparticles as drug delivery carriers for doxorubicin

In the present study, β-cyclodextrin-grafted chitosan nanoparticles (β-CD-g-CS NPs) were prepared using a new ionic gelation strategy involving a synergistic effect of NaCl (150 mmol/L), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, 10 mmol/L), and water bath sonication. This new strategy afforded smaller and more monodisperse β-CD-g-CS NPs vs. the classical ionic gelation method. New HA/β-CD-g-CS NPs were also prepared using the above-mentioned strategy by adding hyaluronic acid (HA) to the β-CD-g-CS copolymer at different weight ratios until the ZP values conversion. The best result was obtained with the weight ratio of w(HA):w(β-CD-g-CS) = 2:1 and furnished new spherical and smooth HA/β-CD-g-CS NPs. Furthermore, the stability of β- CD-g-CS NPs and HA/β-CD-g-CS NPs at 4°C in physiological medium (pH 7.4) was compared for 3 weeks period and showed that HA/β-CD-g-CS NPs were more stable all maintaining their monodispersity and high negative ZP values compared to β-CD-g-CS NPs. Finally, preliminary study of HA/β-CD-g-CS NPs as carrier for the controlled release of the anticancer drug doxorubicin was investigated. These new HA/β-CD-g-CS NPs can potentially be used as drug delivery and targeting systems for cancer treatment.     

Studies of the Local Structure and the g Factors of the Cu<Superscript>2+</Superscript> Site in Y<Subscript>2</Subscript>BaCuO<Subscript>5</Subscript>

The local structure and the g factors (g _x , g _y , and g _z ) of the Cu²⁺ site in Y₂BaCuO₅ are theoretically studied using the perturbation formulas of the g factors for a 3d⁹ ion in orthorhombically elongated octahedra. The orthorhombic field parameters in these formulas are determined from the superposition model and the local geometry of the system. From the calculations, the oxygen octahedron is found to undergo the local elongation ΔZ (≈0.05 Å) along c-axis and the relative bond length variation ΔX (≈0.1 Å) along a- and b-axes, respectively. The calculated g factors based on the above local structure are in good agreement with the experimental data. The relationships between the anisotropies of the g factors and the low symmetrical (orthorhombic) distortions of the Cu²⁺ site in Y₂BaCuO₅ are discussed.     

The Impact of Reduced Gravity on Free Convective Heat Transfer from a Finite,Flat, Vertical Plate

Convective heat transfer is governed by a number of factors including various fluid properties, the presence of a thermal gradient, geometric configuration, flow condition, and gravity. Empirically-derived analytical relationships can be used to estimate convection as a function of these governing parameters. Although it is relatively straightforward to experimentally quantify the contributions of the majority of these variables, it is logistically difficult to assess the influence of reduced-gravity due to practical limitations of establishing this environment. Therefore, in order to explore this regime, a series of tests was conducted to evaluate convection under reduced-gravity conditions averaging 0.45 m/sec² (0.05 g) achieved aboard a parabolic aircraft. The results showed a reduction in net heat transfer of approximately 61% in flight relative to a 1g terrestrial baseline using the same setup. The average experimental Nusselt Number of 19.05 ± 1.41 statistically correlated with the predicted value of 18.90 ± 0.63 (N = 13), estimated using the Churchill-Chu correlation for free convective heat transfer from a finite, flat, vertical plate. Extrapolating this to similar performance in true microgravity (10^?6 g) indicates that these conditions should yield a Nusselt Number of 1.27, which is 2.6% the magnitude of free convection at 1g, or a reduction of 97.4%. With advection essentially eliminated, heat transfer becomes limited to diffusion and radiation, which are gravity-independent and nearly equivalent in magnitude in this case. These results offer a general guideline for integrating components that utilize natural (free) convective gas cooling in a spacecraft habitat and properly sizing the thermal control system.     

On the correspondence from Bayesian log-linear modelling to logistic regression modelling with <Emphasis Type="Italic">g</Emphasis>-priors

Consider a set of categorical variables where at least one of them is binary. The log-linear model that describes the counts in the resulting contingency table implies a specific logistic regression model, with the binary variable as the outcome. Within the Bayesian framework, the g-prior and mixtures of g-priors are commonly assigned to the parameters of a generalized linear model. We prove that assigning a g-prior (or a mixture of g-priors) to the parameters of a certain log-linear model designates a g-prior (or a mixture of g-priors) on the parameters of the corresponding logistic regression. By deriving an asymptotic result, and with numerical illustrations, we demonstrate that when a g-prior is adopted, this correspondence extends to the posterior distribution of the model parameters. Thus, it is valid to translate inferences from fitting a log-linear model to inferences within the logistic regression framework, with regard to the presence of main effects and interaction terms.     

The size and impact of the elite set of publications in scientometric assessments

Several scientometric impact indicators [total citations, h, g, and π-index, percentage rank position (PRP), weighted citation share (WCS)] of 190 elite papers of 15 members of the Hungarian Academy of Sciences active in three different fields were calculated. From the indices the PRP indicator proved to be independent of the citation practices in the fields. The PRP index of a journal paper can be calculated in per cent as unity minus (the rank number of the paper by citation frequency within the publishing journal minus one divided with the total number of papers in the journal) times hundred. The sum of the PRP index of the elite papers of a scientist may characterize his or her total publication performance. The size of the elite set of journal papers within the total was calculated by different methods. The h-index and g-index corresponds to the size of the elite, i.e. number of the elite papers according to the h-statistics and g-statistics, respectively. The number of papers in the π-set is equal to the square root of total papers. The π-index equals to one hundredth of citations to the π-set papers. In the present paper the size of the elite set is determined as the number of papers in the h-set, g-set, or π-set, and as 10 % of total papers, or number of papers cited 2, 3, or 5 times the mean citation rate (MCR) of the publishing journal. The π-citation threshold model is presented for demonstrating how MCR and the distribution of citations over the papers may influence the size of the elite set and the corresponding π-index. It was found that the scientific performances concluded from the π-index obtained from elite sets of different size are in good agreement.