The "gaseous" environment in sealed BRIC-100VC canisters flown on 'Mir' with embryogenic daylily cell cultures.

As part of the "Cellular Mechanisms of Spaceflight-Specific Stress to Plants" experiment, nine BRIC (Biological Research in Canisters) 100VC canisters, each containing four 100 mm dia polycarbonate petri dishes with embryogenic daylily (Hemerocallis sp.) cultures, were launched on 12 Jan 97 (STS-81), transferred to 'Mir' and returned on 24 May 97 (STS-84). Pre-flight, flight and ground control data for temperature, relative humidity, CO2 and ethylene in the BRIC canisters are presented.     

Characterizing parameters of Jatropha curcas cell cultures for microgravity studies

Jatropha (Jatropha curcas) is a tropical perennial species identified as a potential biofuel crop. The oil is of excellent quality and it has been successfully tested as biodiesel and in jet fuel mixes. However, studies on breeding and genetic improvement of jatropha are limited. Space offers a unique environment for experiments aiming at the assessment of mutations and differential gene expression of crops and in vitro cultures of plants are convenient for studies of genetic variation as affected by microgravity. However, before microgravity studies can be successfully performed, pre-flight experiments are necessary to characterize plant material and validate flight hardware environmental conditions. Such preliminary studies set the ground for subsequent spaceflight experiments. The objectives of this study were to compare the in vitro growth of cultures from three explant sources (cotyledon, leaf, and stem sections) of three jatropha accessions (Brazil, India, and Tanzania) outside and inside the petriGAP, a modified group activation pack (GAP) flight hardware to fit petri dishes. In vitro jatropha cell cultures were established in petri dishes containing a modified MS medium and maintained in a plant growth chamber at 25 ± 2 °C in the dark. Parameters evaluated were surface area of the explant tissue (A), fresh weight (FW), and dry weight (DW) for a period of 12 weeks. Growth was observed for cultures from all accessions at week 12, including subsequent plantlet regeneration. For all accessions differences in A, FW and DW were observed for inside vs. outside the PetriGAPs. Growth parameters were affected by accession (genotype), explant type, and environment. The type of explant influenced the type of cell growth and subsequent plantlet regeneration capacity. However, overall cell growth showed no abnormalities. The present study demonstrated that jatropha in vitro cell cultures are suitable for growth inside PetriGAPs for a period of 12 weeks. The parameters evaluated in this study provide the basic ground work and pre-flight assessment needed to justify a model for microgravity studies with jatropha in vitro cell cultures. Future studies should focus on results of experiments performed with jatropha in vitro cultures in microgravity.     

Early development of fern gametophytes in microgravity.

Dormant spores of the fern Ceratopteris richardii were flown on Shuttle mission STS-93 to evaluate the effects of micro-g on their development and on their pattern of gene expression. Prior to flight the spores were sterilized and sown into one of two environments: (1) Microscope slides in a video-microscopy module; and (2) Petri dishes. All spores were then stored in darkness until use. Spore germination was initiated on orbit after exposure to light. For the spores on microscope slides, cell level changes were recorded through the clear spore coat of the spores by video microscopy. After their exposure to light, spores in petri dishes were frozen in orbit at four different time points during which on earth gravity fixes the polarity of their development. Spores were then stored frozen in Biological Research in Canister units until recovery on earth. The RNAs from these cells and from 1-g control cells were extracted and analyzed on earth after flight to assay changes in gene expression. Video microscopy results revealed that the germinated spores developed normally in microgravity, although the polarity of their development, which is guided by gravity on earth, was random in space. Differential Display-PCR analyses of RNA extracted from space-flown cells showed that there was about a 5% change in the pattern of gene expression between cells developing in micro-g compared to those developing on earth.     

Improvements in the re-flight of spaceflight experiments on plant tropisms

In order to effectively study phototropism, the directed growth in response to light, we performed a series of experiments in microgravity to better understand light response without the “complications” of a 1-g stimulus. These experiments were named TROPI (for tropisms) and were performed on the European Modular Cultivation System (EMCS), a laboratory facility on the International Space Station (ISS). TROPI-1 was performed in 2006, and while it was a successful experiment, there were a number of technical difficulties. We had the opportunity to perform TROPI-2 in 2010 and were able to optimize experimental conditions as well as to extend the studies of phototropism to fractional gravity created by the EMCS centrifuge. This paper focuses on how the technical improvements in TROPI-2 allowed for a better experiment with increased scientific return. Major modifications in TROPI-2 compared to TROPI-1 included the use of spaceflight hardware that was off-gassed for a longer period and reduced seed storage (less than 2 months) in hardware. These changes resulted in increased seed germination and more vigorous growth of seedlings. While phototropism in response to red illumination was observed in hypocotyls of seedlings grown in microgravity during TROPI-1, there was a greater magnitude of red-light-based phototropic curvature in TROPI-2. Direct downlinking of digital images from the ISS in TROPI-2, rather than the use of analog tapes in TROPI-1, resulted in better quality images and simplified data analyses. In TROPI-2, improved cryo-procedures and the use of the GLACIER freezer during transport of samples back to Earth maintained the low temperature necessary to obtain good-quality RNA required for use in gene profiling studies.     

Optimization of moisture content for wheat seedling germination in a cellulose acetate medium for a space flight experiment.

The Porous Tube Plant Nutrient Delivery System (PTPNDS), a hydrophilic, microporous ceramic tube hydroponic system designed for microgravity, will be tested in a middeck locker of the Space Shuttle. The flight experiment will focus on hardware operation and assess its ability to support seed germination and early seedling growth in microgravity. The water controlling system of the PTPNDS hardware has been successfully tested during the parabolic flight of the KC-135. One challenge to the development of the space flight experiment was to devise a method of holding seeds to the cylindrical porous tube. The seed-holder must provide water and air to the seed, absorb water from the porous tube, withstand sterilization, provide a clear path for shoots and roots to emerge, and be composed of flight qualified materials. In preparation for the flight experiment, a wheat seed-holder has been designed that utilizes a cellulose acetate plug to facilitate imbibition and to hold the wheat seeds in contact with the porous tube in the correct orientation during the vibration of launch and the microgravity environment of orbit. Germination and growth studies with wheat at a range of temperatures showed that optimal moisture was 78% (by weight) in the cellulose acetate seed holders. These and other design considerations are discussed.     

Liquid motion in partially filled containers: Preliminary results of the D-1 mission

Experiment WL-FPM-08, or also: FLIM, has been executed in the FPM on the D-1 mission. The experiment is a continuation of an earlier one on Spacelab-1, and seeks to study capillary liquid flow in transparent models of spacecraft tanks. A typical dimension in a container is five centimeters.Six experiment runs have been conducted with liquid in a straight cylinder container with hemispherical ends. The flight results are recorded on cine film that is processed for photogrammetric data. The liquid, water, remained at one end of the container throughout the experiments and showed a stuck contact line with the wall (perspex). Since the liquid contained bubbles and the contact line was non-circular, interpretation of the results is not straightforward.For axisymmetric configurations a computer program, SAVOF, has been developed at NLR for numerical simulations of the liquid flow. The dependence of the lowest axisymmetric resonance of the liquid on the contact line (stuck) position has been calculated both for a non-rotating and a rotating tank. The flight results are generally in line with the numerical results.     

Recent advances in technologies required for a "Salad Machine".

Future long duration, manned space flight missions will require life support systems that minimize resupply requirements and ultimately approach self-sufficiency in space. Bioregenerative life support systems are a promising approach, but they are far from mature. Early in the development of the NASA Controlled Ecological Life Support System Program, the idea of onboard cultivation of salad-type vegetables for crew consumption was proposed as a first step away from the total reliance on resupply for food in space. Since that time, significant advances in space-based plant growth hardware have occurred, and considerable flight experience has been gained. This paper revisits the "Salad Machine" concept and describes recent developments in subsystem technologies for both plant root and shoot environments that are directly relevant to the development of such a facility.     

Life sciences flight hardware development for the International Space Station.

During the construction phase of the International Space Station (ISS), early flight opportunities have been identified (including designated Utilization Flights, UF) on which early science experiments may be performed. The focus of NASA's and other agencies' biological studies on the early flight opportunities is cell and molecular biology; with UF-1 scheduled to fly in fall 2001, followed by flights 8A and UF-3. Specific hardware is being developed to verify design concepts, e.g., the Avian Development Facility for incubation of small eggs and the Biomass Production System for plant cultivation. Other hardware concepts will utilize those early research opportunities onboard the ISS, e.g., an Incubator for sample cultivation, the European Modular Cultivation System for research with small plant systems, an Insect Habitat for support of insect species. Following the first Utilization Flights, additional equipment will be transported to the ISS to expand research opportunities and capabilities, e.g., a Cell Culture Unit, the Advanced Animal Habitat for rodents, an Aquatic Facility to support small fish and aquatic specimens, a Plant Research Unit for plant cultivation, and a specialized Egg Incubator for developmental biology studies. Host systems (Figure 1A, B: see text), e.g., a 2.5 m Centrifuge Rotor (g-levels from 0.01-g to 2-g) for direct comparisons between g and selectable g levels, the Life Sciences Glovebox for contained manipulations, and Habitat Holding Racks (Figure 1B: see text) will provide electrical power, communication links, and cooling to the habitats. Habitats will provide food, water, light, air and waste management as well as humidity and temperature control for a variety of research organisms. Operators on Earth and the crew on the ISS will be able to send commands to the laboratory equipment to monitor and control the environmental and experimental parameters inside specific habitats. Common laboratory equipment such as microscopes, cryo freezers, radiation dosimeters, and mass measurement devices are also currently in design stages by NASA and the ISS international partners.     

Influence of microgravity on mitogen binding and cytoskeleton in Jurkat cells.

The effects of microgravity on Jurkat cells--a T-lymphoid cell line--was studied on a sounding rocket flight. An automated pre-programmed instrument permitted the injection of fluorescent labelled concanavalin A (Con A), culture medium and/or fixative at given times. An in-flight 1 g centrifuge allowed the comparison of the data obtained in microgravity with a 1 g control having the same history related to launch and re-entry. After flight, the cells fixed either at the onset of microgravity or after a or 12 minute incubation time with fluorescent concanavalin A were labelled for vimentin and actin and analysed by fluorescence microscopy. Binding of Con A to Jurkat cells is not influenced by microgravity, whereas patching of the Con A receptors is significantly lower. A significant higher number of cells show changes in the structure of vimentin in microgravity. Most evident is the appearance of large bundles, significantly increased in the microgravity samples. No changes are found in the structure of actin and in the colocalisation of actin on the inner side of the cell membrane with the Con A receptors after binding of the mitogen.     

火箭空中爆炸冲击波峰值超压预测方法

研究火箭空中爆炸冲击波参数预测方法对于乘员舱的安全评估具有重要意义。为了探究火箭空中爆炸时飞行高度对峰值超压的影响,获取冲击波参数快速预测方法,利用ANSYS/LS-DYNA有限元软件对火箭飞行至0~20 km高度爆炸进行了有限元仿真分析。结果表明,作用于乘员舱的冲击波峰值超压随飞行高度的增加而快速减小。火箭空中爆炸冲击波压强衰减系数与飞行高度之间的关系服从二次函数衰减。在此基础上,提出了考虑高度效应的火箭空中爆炸冲击波峰值超压预测公式,可为乘员舱的快速危害性评估以及防护研究提供一定参考。      

Changes in developmental capacity of artemia cyst and chromosomal aberrations in lettuce seeds flown aboard Salyut-7 (Biobloc III experiment).

This paper gives the results of investigations performed on the first container (A) of the Biobloc III experiment, flown aboard the orbital station Salyut 7 for 40 days. The space flight resulted in a decreased developmental capacity of Arterlia cysts, hit or not hit by the HZE particles. No effect was observed in cysts in bulk. A synergetic effect of microgravity and gamma pre irradiation is described. The germination of in-flight lettuce seeds was decreased. The space flight resulted also in a higher percentage of cells with chromosomal aberrations. Relations between biological response, TEL and location of HZE particles are discussed.     

Estimation of a cosmonaut's radiation hazard during long-term space missions on the basis of a generalized dosimetric function.

This paper presents a new concept of radiation hazard assessment for spacecraft crew members during long term space missions on the basis of a generalized dosimetric function. This new dosimetric function enables a complicated nature of space radiation exposure to be reduced to the conditions of a standard irradiation. It can be obtained on the basis of mean-tissue equivalent dose values calculated for each space radiation source and transmission coefficients describing the influence of the complex spatial and temporal distribution of the absorbed dose in the cosmonaut's body on the radiobiological effects. The combination of cosmic ionizing radiation with other non-radiation nature factors in flight can also be accounted for. In terms of the generalized dose, it is possible to assess the nature and extent of lowering a crew working capacity, as well as radiation risk, both during a flight and post flight period.     

Optical and electron-microscopic studies of the Funaria hygrometrica protonema after cultivation for 96 days in space

$\text{Funaria hygrometrica}$ protonema cells grown in the “IFS-2” (Inoculating fixing system) for 96 days on board the Salyut 6 — Soyuz 32 orbital scientific station were examined by light and electron-microscopy. Investigation of experimental and control cells of the moss protonema showed common features as well as distinctions in their structure. Protonema cells of $\text{Funaria hygrometrica}$ both differentiate and undergo photosynthesis during space flight. Changes in cell shape, decreased cell size, a reduction in the volume of starch granules, and altered chloroplast structure were observed.     

First flight of the ASTROCULTURE (TM) experiment as a part of the U.S. Shuttle/MIR program.

A number of space-based experiments have been conducted to assess the impact of microgravity on plant growth and development. In general, these experiments did not identify any profound impact of microgravity on plant growth and development, though investigations to study seed development have indicated difficulty in plants completing their reproductive cycle. However, it was not clear whether the lack of seed production was due to gravity effects or some other environmental condition prevailing in the unit used for conducting the experiment. The ASTROCULTURE (TM) flight unit contains a totally enclosed plant chamber in which all the critically important environmental conditions are controlled. Normal wheat (Triticum aestivum L.) growth and development in the ASTROCULTURE (TM) flight unit was observed during a ground experiment conducted prior to the space experiment. Subsequent to the ground experiment, the flight unit was transported to MIR by STS-89, as part of the U.S. Shuttle/MIR program, in an attempt to determine if super dwarf wheat plants that were germinated in microgravity would grow normally and produce seeds. The experiment was initiated on-orbit after the flight unit was transferred from the Space Shuttle to MIR. The ASTROCULTURE (TM) flight unit performed nominally for the first 24 hours after the flight unit was activated, and then the unit stopped functioning abruptly. Since it was not possible to return the unit to nominal operation it was decided to terminate the experiment. On return of the flight unit, it was confirmed that the control computer of the ASTROCULTURE (TM) flight unit sustained a radiation hit that affected the control software embedded in the computer. This experience points out that at high orbital inclinations, such as that of MIR and that projected for the International Space Station, the danger of encountering harmful radiation effects are likely unless the electronic components of the flight hardware are resistant to such impacts.     

Evaluating and optimizing horticultural regimes in space plant growth facilities.

In designing innovative space plant growth facilities (SPGF) for long duration space flight, various limitations must be addressed including onboard resources: volume, energy consumption, heat transfer and crew labor expenditure. The required accuracy in evaluating on board resources by using the equivalent mass methodology and applying it to the design of such facilities is not precise. This is due to the uncertainty of the structure and not completely understanding the properties of all associated hardware, including the technology in these systems. We present a simple criteria of optimization for horticultural regimes in SPGF: Qmax = max [M x (EBI)2/(V x E x T], where M is the crop harvest in terms of total dry biomass in the plant growth system; EBI is the edible biomass index (harvest index), V is volume occupied by the crop; E is the crop light energy supply during growth; T is the crop growth duration. The criterion reflects directly on the consumption of onboard resources for crop production.     

Un dispositif experimental utilisant des ballons plafonnants pour l'etude de la couche limite atmospherique

This paper presents a new system of constant volume balloons used to study the dynamical and thermodynamical properties of the tropospheric boundary layer. The system allows to simultaneously localize up to 30 balloons and to observe thermodynamical characteristics of the air during their flight inside the boundary layer. Each balloon is equipped with a radar reflector and a sounding system giving information each second on pressure, humidity and temperature by sequential emissions which are received at a ground station. The trajectories of balloons are obtained by a non-tracking S-band radar where a special hardware processing unit allows to real time cancellation of most ground clutter. A complete set of the balloons positions is obtained every two or three minutes.The system was tested in October 1984 during a field experiment in the south of France. Results of the experimental procedure and of the quality of the balloon, radar and sounding capabilities are given. The scientific use of constant volume balloons in order to study the atmospheric boundary layer is examined.