Columbus, the European Physiology Modules Facility and CADMOS

Columbus, the European Space Agency (ESA) orbital facility laboratory will be launched in December 2007 and attached to the International Space Station (ISS). In its launch configuration, Columbus includes 4 multi-user facilities: one of them is the European Physiology Modules Facility, also called EPM. The EPM will be devoted to Human Physiology; it will be collocated in the Columbus module with two other physiology racks, i.e. the HRF-1 and HRF-2 American racks (Human Research Facility). CADMOS is part of the French Space Agency, located in Toulouse; it has been designated by the European Space Agency as the Facility Responsible Centre (FRC) for the EPM. As a User Support and Operations Centre, CADMOS main tasks are to help the scientists to prepare and perform their experiments in Space and to monitor operations on the ISS.     

The Biotechnology Facility for International Space Station

The primary mission of the Cellular Biotechnology Program is to advance microgravity as a tool in basic and applied cell biology. The microgravity environment can be used to study fundamental principles of cell biology and to achieve specific applications such as tissue engineering. The Biotechnology Facility (BTF) will provide a state-of-the-art facility to perform cellular biotechnology research onboard the International Space Station (ISS). The BTF will support continuous operation, which will allow performance of long-duration experiments and will significantly increase the on-orbit science throughput.     

Mice Drawer System: phase c/d development and perspective

MDS (Mice Drawer System) is the Facility that will support the research on board the International Space Station (ISS). Funded by ASI with Laben as industrial Prime Contractor, MDS will also permit its utilisation for other research programs with mice. The most attractive feature of MDS is that six mice can he kept separate inside dedicated and isolated cages. This permits to perform six experiments in parallel, one for each mouse. In the first utilisation flight of MDS facility the selected experiment is devoted to study human bone formation and specific countermeasures to prevent osteoporosis using mice under microgravity conditions as a model.     

The crystallization of biological macromolecules under microgravity: a way to more accurate three-dimensional structures?

The crystallization of proteins and other biological particles (including nucleic acids, nucleo-protein complexes and large assemblies such as nucleosomes, ribosomal subunits or viruses) in a microgravity environment can produce crystals having lesser defects than crystals prepared under normal gravity on earth. Such microgravity-grown crystals can diffract X-rays to a higher resolution and have a lower mosaic spread. The inferred electron density maps can be richer in details owing to which more accurate three-dimensional structure models can be built. Major results reported in this field of research are reviewed. Novel ones obtained with the Advanced Protein Crystallization Facility are presented. For structural biology, practical applications and implications associated with crystallization and crystallography onboard the International Space Station are discussed.     

Influence of beta-radiation on human erythrocyte surface potential under eicosanoid metabolism inhibition

The aim of the research consisted in the study of influence of beta-radiation on response of erythrocyte surface potential to inhibitors of eicosanoid metabolism enzymes (cyclo-, lipoxygenase and phospholipase A2). It was shown, that inhibitors of phospholipase A2 (quinacrine, 10-100 microM), cyclooxygenase (aspirin, 10-100 microM) and cyclo- and lipoxygenase (BW755c, 1-100 microM) lowered electrophoretic mobility (EPM) of erythrocytes by 20-30%. An analogous effect can be exerted by beta-radiation. Nonradioactive leucine in the studied concentrations cannot simulate EPM erythrocytes. Response of cellular EPM to these inhibitors depended on their concentration in the incubation medium. Addition of 14C to the incubation medium changed response of EPM of cells to inhibitors of cyclo- and lipoxygenase but not to quinacrine. However beta-radiation fully abolished the stimulative action of nonspecific activator of phospholipase A2 (Ca-independent), H2O2, on cellular EPM. Under these conditions beta-radiation enhanced EPM response to aspirin only at concentration of 100 microM. The EPM response to BW755c is reduced by irradiation at all concentrations with the exception of equal-effective one (10 microM). Data obtained evidence for modification of eicosanoid metabolism by beta-radiation, probably, as a result of phospholipase A2 inhibition, as evident from elimination by radiation of stimulated action of hydrogen peroxide on EPM. The radiation action can also affect the cyclooxygenase lipoxygenase activity ratio, this activity being mediated by cellular membrane signaling systems.     

Developmental status of Aquatic Animal Experiment Facility, Aquatic Habitat (AQH), for International Space Station.

Mitsubishi Heavy Industries (MHI) and Japan Aerospace Exploration Agency (JAXA) have been studying Aquatic Animal Experiment Facility, Aquatic Habitat (AQH), for International Space Station (ISS). The AQH will have the capabilities to accommodate small freshwater fish and amphibian for maximum 90 days on orbit. Three-generations of small freshwater fish (medaka and zebrafish), and egg through metamorphosis of amphibian (African clawed toad) could be experimented by AQH. Various experimental functions such as automatic feeding, air-water interface, day/night cycle, video observation, and specimen sampling mechanism will be also equipped in AQH. The water circulation system was improved from the past aquatic facilities for Space Shuttle experiments under the consideration of the long life-time, and a brand-new specimen chamber was developed to equip the above various experimental functions. Currently the prototype model of water circulation system and specimen chambers have been manufactured and biological compatibility tests are being conducted with medaka. The current developmental status of AQH is summarized.     

Usefulness of the Centrifuge Accommodation Module for analyzing gravity responses in plant seedlings]

Onboard centrifuges are indispensable tools for clarifying the effects of microgravity on various physiological processes in plant seedlings. Centrifuges are basically attached to the incubators designed for the International Space Station (ISS). However, because of the limitation in size, that loaded to the Cell Biology Experiment Facility (CBEF) is usable only to some small seedlings such as Arabidopsis. The Centrifuge Accommodation Module (CAM) has great advantages in the size and the amounts of plant materials feasible to load, the quality of acceleration produced, and the easiness of operation on it. The CAM is an apparatus that characterizes the ISS most and its construction on schedule is highly expected.     

One-trial tolerance to the effects of chlordiazepoxide in the elevated plus-maze is not due to acquisition of a phobic avoidance of open arms during initial exposure

A single exposure to the elevated plus-maze (EPM) test of anxiety reduces or abolishes the anxiolytic-like efficacy of benzodiazepines. This phenomenon called one-trial tolerance has been suggested to represent the acquisition of a phobic-like response to the open arms during trial 1. The present study was designed to examine the effects of chlordiazepoxide (5 mg/kg, ip) on the behaviour of rats in a conventional EPM apparatus after previous exposure to a four-open-arm EPM, a four-enclosed arm EPM or a conventional EPM, as well as in naive rats. Chlordiazepoxide had clear-cut anxiolytic-like effects (increased percentage of time spent on the open arms) in a traditional EPM in naive rats and in animals previously exposed to a four-open-arm EPM. However, it was ineffective in rats previously exposed to a traditional or a four-closed-arm EPM. Thus, the phenomenon of one-trial tolerance does not depend upon initial open-arm experience.     

Availability and Readability of Emergency Preparedness Materials for Deaf and Hard-of-Hearing and Older Adult Populations: Issues and Assessments

A major public health challenge is to communicate effectively with vulnerable populations about preparing for disasters and other health emergencies. People who are Deaf or Hard of Hearing (Deaf/HH) and older adults are particularly vulnerable during health emergencies and require communications that are accessible and understandable. Although health literacy studies indicate that the readability of health communication materials often exceeds people’s literacy levels, we could find no research about the readability of emergency preparedness materials (EPM) intended for Deaf/HH and older adult populations. The objective of this study was to explore issues related to EPM for Deaf/HH and older adult populations, to assess the availability and readability of materials for these populations, and to recommend improvements. In two California counties, we interviewed staff at 14 community-based organizations (CBOs) serving Deaf/HH clients and 20 CBOs serving older adults selected from a stratified, random sample of 227 CBOs. We collected 40 EPM from 10 CBOs and 2 public health departments and 40 EPM from 14 local and national websites with EPM for the public. We used computerized assessments to test the U.S. grade reading levels of the 16 eligible CBO and health department EPM, and the 18 eligible website materials. Results showed that less than half of CBOs had EPM for their clients. All EPM intended for clients of Deaf/HH-serving CBOs tested above the recommended 4^th grade reading level, and 91% of the materials intended for clients of older adult-serving CBOs scored above the recommended 6^th grade level. EPM for these populations should be widely available through CBOs and public health departments, adhere to health literacy principles, and be accessible in alternative formats including American Sign Language. Developers should engage the intended users of EPM as co-designers and testers. This study adds to the limited literature about EPM for these populations.     

Developmental testing of the Advanced Animal Habitat to determine compatibility with rats and mice

The Research Animal Holding Facility (RAHF) and the Animal Enclosure Module (AEM) have housed rats during Space Shuttle flights since the 1980s, but the operational constraints of the hardware have limited the scientific return from these Shuttle flights. The RAHF provides environmental control and monitoring for 24 rats with in-flight animal access, but it must be flown in the Spacelab. Due to the infrequent availability of Spacelab flights, rodent experiments rely heavily on the AEM. Unfortunately, the AEM supports only six rats, has no environmental control and provides no animal access in flight. The Advanced Animal Habitat (AAH) is being developed to support up to 12 adult rats or 30 adult mice for up to 30 days, provide active temperature control, animal telemetry and on-orbit video, record environmental parameters in the animal cage, and provide in-flight animal access in the Middeck, the Spacelab or the Space Station. To ensure the AAH can meet these requirements, animal testing is being conducted with rats and mice in every step of development. Testing began with the cage configuration.     

Effect of ultra-low dose of ionizing radiation on the surface potential of erythrocytes

Effect of ionizing radiation in ultralow dose (5 microGy) on responses of erythrocyte electrophoretic motility (EPM) as a result of adrenoreceptor ligands binding (0.01-100 microM) has been investigated. The opposite directional EPM responses to agonists (adrenaline, isoprenaline) and antagonist (propranolol) of beta-adrenoreceptors was shown. At that, EPM response to the radiation coincides both with the direction and value of acting the beta-adrenoreceptor agonists depressing EPM. The EPM response to a combined action of beta-adrenoreceptors antagonist, propranolol (10 microM), and ionizing radiation is additive. The above listed is capable to evidence about the essential role of adrenoreceptors at formation of erythrocyte membrane surface charge under action of ionizing radiation in ultralow doses.     

The effect of temperature on the survival of microorganisms in a deep space vacuum

A Space Molecular Sink Research Facility (Molsink) was used to evaluate the ability of microorganisms to survive the vacuum of outer space. this facility could be programmed to simulate flight spacecraft vacuum environments at pressures in the 10^–10 torr range and thermal gradients (30 to 60°C) closely associated to surface temperatures of inflight spacecraft.Initial populations ofStaphylococcus epidermidis and aMicrococcus sp. were reduced approximately 1 log while exposed to –105 and 34°C, and approximately 2 logs while exposed to 59°C for 14 days in the vacuum environment.Spores ofBacillus subtilis var.niger were less affected by the environment. Initial spore populations were reduced 0.2, 0.3, and 0.8 log during the 14-day vacuum exposure at –124, 34, and 59°C, respectively.This paper presents the results of one phase of research carried out at the Jet Propulsion Laboratory, California Institute of Technology, under Contract No. NAS 7-100, sponsored by the National Aeronautics and Space Administration.     

The electrophoretic mobility of gram-negative and gram-positive bacteria: an electrokinetic analysis.

The electrophoretic mobility (EPM) of a variety of Gram-negative and Gram-positive bacteria was measured with a Penkem S3000 analyser. Under standard growth conditions and neutral pH all cells displayed a negative EPM. The polysaccharide capsules of Escherichia coli strains K1, K5, K29 and K30 generated the highest EPM; to a lesser and varying degree O-antigens with charged groups and core lipopolysaccharides also contribute to the net EPM. Very little negative EPM was measured in suspension cultures of the gliding bacterium Cytophaga U67. No difference in the EPM was observed between rapidly growing and stationary-phase E. coli B. De-energization of the cell membranes by carbonyl cyanide m-chlorophenylhydrazone (CCCP) did not affect the EPM of wild-type and deep rough mutants of E. coli; and the EPM of Cytophaga U67 and Acholeplasma laidlawii remained unaltered by CCCP when measured in their respective growth media. Extrusion of filamentous bacteriophage f1 from cells of its host, E. coli A95, caused a shift to a higher negative EPM. We also measured a variety of Gram-positive strains, all of which displayed different EPMs. When membrane fractions of E. coli were adsorbed to latex spheres, characteristic differences between the EPM of beads coated with either inner or outer membrane were observed. The results suggest that the rapid EPM analysis is a useful tool to study the net electric charge of microorganisms and to examine changes of surface properties during interaction of cells with viruses, proteins (antibody) and charged antibiotics.     

Increases in plasma corticosterone and stretched-attend postures in rats naive and previously exposed to the elevated plus-maze are sensitive to the anxiolytic-like effects of midazolam

A single exposure to the elevated plus-maze test (EPM) reduces or abolishes the anxiolytic efficacy of benzodiazepines on a second trial. This phenomenon known as one-trial tolerance (OTT) is considered to be due to a shift in the emotional state of the animals across the test/retest sessions. Activation of the hypothalamic-pituitary-adrenocortical (HPA) axis has been considered to be an adaptive response to stressful or challenging situations such as height and openness of the EPM. This work looks at the phenomenon of OTT to the benzodiazepine compound midazolam through the conjoint examination of the novel ethological measures of the EPM and adrenocortical response of rats exposed to single and repeated sessions of the EPM. The results obtained confirmed that the approach/avoidance conflict on the first trial of the EPM is very sensitive to the anxiolytic effects of benzodiazepines. Moreover, stressful stimuli present upon initial exposure to the EPM render the standard measures of the EPM resistant to the anxiolytic effects of benzodiazepines on retest. However, the increases in plasma corticosterone and in risk assessment behavior observed in rats submitted to single or repeated sessions in the EPM were reversed by pretreatment with midazolam. The administration of metyrapone, a glucocorticoid synthesis blocker, decreased risk assessment but did not affect locomotion and anxiety-like behaviors. It is suggested that the detection of the dangerous environment through the stretched-attend postures in the second trial leads to the known low level of exploration and the consequent OTT upon retest. Moreover, in view of the similarity between the risk assessment and plasma corticosterone patterns in both naive and experienced rats, this hormone-behavior profile may be crucial for the expression of OTT to benzodiazepines in rodents exposed to the EPM.     

Calcium-dependent process in reduction of cell surface charge after x-irradiation.

The electrophoretic mobility (EPM) of rat erythrocytes and cultured melanoma cells decreased with time after X-irradiation in the presence of calcium at concentrations higher than 10 (-5) M. At 37 degrees C, the presence of calcium for the first 20 min of exposure was suffcient to induce the EPM reduction, and Ca 2+ administration subsequent to Ca 2+ -free incubation for 30 min following irradiation had no effect on EPM. At lower temperatures, from 10 down to 20 degrees C however, the effect of calcium on the reduction of EPM decreased drastically. If the cells were kept Ca 2+ -inonophore A23187 also induced to decrease in EPM only in the presence of Ca 2+. These results revealed the transitory existence of membrane condition reactive to extracellular Ca 2+ immediately after X-irradiation, which can be postponed at low temperatures. The reduction of EPM by Ca 2+ -ionophore might suggest that the influx of Ca 2+ is a step in the reduction of EPM after X-irradiation.     

Protein and virus crystal growth on international microgravity laboratory-2.

Two T = 1 and one T = 3 plant viruses, along with a protein, were crystallized in microgravity during the International Microgravity Laboratory-2 (IML-2) mission in July of 1994. The method used was liquid-liquid diffusion in the European Space Agency's Advanced Protein Crystallization Facility (APCF). Distinctive alterations in the habits of Turnip Yellow Mosaic Virus (TYMV) crystals and hexagonal canavalin crystals were observed. Crystals of cubic Satellite Tobacco Mosaic Virus (STMV) more than 30 times the volume of crystals grown in the laboratory were produced in microgravity. X-ray diffraction analysis demonstrated that both crystal forms of canavalin and the cubic STMV crystals diffracted to significantly higher resolution and had superior diffraction properties as judged by relative Wilson plots. It is postulated that the establishment of quasi-stable depletion zones around crystals growing in microgravity are responsible for self-regulated and more ordered growth.     

Agro-biology for bioregenerative Life Support Systems in long-term Space missions: General constraints and the Italian efforts

Abstract

Future Space stations and Space habitats/outposts should be envisioned as self-sufficient ecological closed or semi-closed systems. The Italian Space Agency (ASI) projects have involved many research groups from complementary areas with a final goal of designing a facility for plant cultivation in Space. It has been critical to (i) identify, for this particular environment, highly productive species able to optimize O₂ production/CO₂ consumption, and (ii) develop high-tech controlled environments. Research activities have included seedling production in simulated microgravity and in Space with the two-fold objective of (i) integrating the crew diet with fresh food, and (ii) studying specific biological phenomena. Another research topic concerned pollen biology as a critical component for seed-to-seed cycles but also for gametophytic selection. In this context, a review of the main scientific topics on plant Space biology and of the Italian efforts on agro-biology for bioregenerative Life Support Systems will be presented and discussed in this paper.     

Epimorphin Regulates Bile Duct Formation via Effects on Mitosis Orientation in Rat Liver Epithelial Stem-Like Cells

Understanding how hepatic precursor cells can generate differentiated bile ducts is crucial for studies on epithelial morphogenesis and for development of cell therapies for hepatobiliary diseases. Epimorphin (EPM) is a key morphogen for duct morphogenesis in various epithelial organs. The role of EPM in bile duct formation (DF) from hepatic precursor cells, however, is not known. To address this issue, we used WB-F344 rat epithelial stem-like cells as model for bile duct formation. A micropattern and a uniaxial static stretch device was used to investigate the effects of EPM and stress fiber bundles on the mitosis orientation (MO) of WB cells. Immunohistochemistry of liver tissue sections demonstrated high EPM expression around bile ducts in vivo. In vitro, recombinant EPM selectively induced DF through upregulation of CK19 expression and suppression of HNF3α and HNF6, with no effects on other hepatocytic genes investigated. Our data provide evidence that EPM guides MO of WB-F344 cells via effects on stress fiber bundles and focal adhesion assembly, as supported by blockade EPM, β1 integrin, and F-actin assembly. These blockers can also inhibit EPM-induced DF. These results demonstrate a new biophysical action of EPM in bile duct formation, during which determination of MO plays a crucial role.     

Microbial characterization of the Mars Odyssey spacecraft and its encapsulation facility

Microbial characterization of the Mars Odyssey spacecraft and the Kennedy Space Center Spacecraft Assembly and Encapsulation Facility II (SAEF-II) was carried out by both culture-based and molecular methods. The most dominant cultivable microbes were species of Bacillus, with comamonads, microbacteria and actinomycetales also represented. Several spore-forming isolates were resistant to gamma-radiation, UV, H2O2 and desiccation, and one Acinetobacter radioresistens isolate and several Aureobasidium, isolated directly from the spacecraft, survived various conditions. Sequences arising in clone libraries were fairly consistent between the spacecraft and facility; predominant genera included Variovorax, Ralstonia and Aquaspirillum. This study improves our understanding of the microbial community structure, diversity and survival capabilities of microbes in an encapsulation facility and physically associated with colocated spacecraft.     

CHANGES IN THE ELECTROPHORETIC MOBILITY OF PRESUMPTIVE ECTODERM CELLS TREATED WITH MESODERMAL INDUCING SUBSTANCES

Using gastrula ectoderm of Cynops pyrrhogaster , the electrophoretic mobility (EPM) of cells was measured. Before EPM measurement, the cells were treated with either LiCl or swimbladder tissue, which are known to possess endo- and mesodermal inducing activity. The EPM of the treated cells increased after 48 hr. A significant difference was observed between the control and experimental series. In addition to the measurements of EPM, histological sections of the explants were examined. Before obvious tissue differentiation occurred in the explants, definitive changes in the EPM values were recorded at 48 and 72 hr after treatment.