The Student Spaceflight Experiments Program (SSEP) is a United States national science, technology, engineering, and mathematics initiative that aims to increase student interest in science by offering opportunities to perform spaceflight experiments. The experiment detailed here was selected and flown aboard the third SSEP mission and the first SSEP mission to the International Space Station (ISS). Caenorhabditis elegans is a small, transparent, self-fertilizing hermaphroditic roundworm that is commonly used in biological experiments both on Earth and in Low Earth Orbit. Past experiments have found decreased expression of mRNA for several genes whose expression can be controlled by the FOXO transcription factor DAF-16. We flew a daf-16 mutant and control worms to determine if the effects of spaceflight on C. elegans are mediated by DAF-16. The experiment used a Type Two Fluids Mixing Enclosure (FME), developed by Nanoracks LLC, and was delivered to the ISS aboard the SpaceX Dragon and returned aboard the Russian Soyuz. The short time interval between experiment selection and the flight rendered preflight experiment verification tests impossible. In addition, published research regarding the viability of the FME in life science experiments was not available. The experiment was therefore structured in such a way as to gather the needed data. Here we report that C. elegans can survive relatively short storage and activation in the FME but cannot produce viable populations for post-flight analysis on extended missions. The FME appears to support short-duration life science experiments, potentially on supply or crew exchange missions, but not on longer ISS expeditions. Additionally, the flown FME was not properly activated, reportedly due to a flaw in training procedures. We suggest that a modified transparent FME could prevent similar failures in future flight experiments.
Research Containing: Biology
Bone Proteomics experiment (BOP): the first proteomic analysis of mammalian cells cultured in weightlessness conditions
Purpose. Bone mass loss is a major consequence of extended periods of weightlessness. Many studies performed on astronauts and animals have shown that impaired maturation of osteoblast cells as well as a decrease of their bone-synthesising activity play key roles in microgravity-dependent bone mass loss. Several experiments on single cells and tissues showed that weightlessness can also influence cells cultured in vitro. Many molecular mechanisms are affected, among which the cytoskeleton, signal transduction cascades and gene expression. However, the underlying mechanisms of these changes and their molecular consequences are far from being fully understood. In contrast to weightlessness, dynamic mechanical loading increases bone density and strength and promotes osteoblast proliferation, differentiation and matrix production. A growing body of evidence points to extracellular nucleotides (i.e. ATP and UTP) as soluble factors that are released by several cell types in response to mechanical stimulation and that eventually trigger an intracellular signal. We have recently demonstrated that ATP and UTP, as well as mechanical stimulation, can activate two fundamental transcription factors, Runx2 and Egr-1, in the human HOBIT osteoblast cell line [Pines A. et al.,Biochem J. 2003; Costessi A. et al., Bone, 2005]. The purpose of the present study was to investigate the possible role(s) of extracellular nucleotides in the molecular response of osteoblast cells to weightlessness conditions. We focused on two aspects: 1. whether administration of ATP could stimulate osteoblast cells in weightlessness, possibly balancing or overcoming its known negative effects; 2. An analysis of the proteome of osteoblast cells exposed to weightlessness by means of two dimension electrophoresis (2-DE) coupled to mass spectrometry, to identify new molecular targets. Methodology – The BOP experiment. BOP was selected in the Success 2002 Student Contest organized by ESA and awarded to A.C. We developed and produced a new and low-cost dedicated hardware to support the experiment. We decided to use the human osteoblast cell line MG-63, since they had been studied in previous space experiments. The cells were grown in space for about five days in three chambers (control cells and cells treated with ATP for 20’ or 3 hours) and eventually lysed. A parallel ground experiment was performed. BOP flew during the Italian Soyuz Taxi flight in April 2005. Results and conclusions. We developed in less than nine months a new hardware that provided 70 cm2 per chamber and a total of more than 200 cm2. Preliminary analysis indicate that administration of ATP to MG-63 cells cultured in weightlessness conditions is able to increase ERK phosphorylation. Analysis of 2D gels revealed several differentially regulated proteins in response to ATP treatment. The identification of these proteins is in progress. To the best of our knowledge, BOP is the first proteomic study on mammalian cells cultured in space. The conclusion of the analysis will reveal new aspects of osteoblast biology and provide new insights into the molecular responses of human cells to weightlessness.
Two experiments were conducted aboard the International Space Station (ISS) in 2008 and 2009 that engaged elementary and middle school teachers and students worldwide in authentic science investigations designed to increase student knowledge of, and interest in biology and space life science studies and biomedical careers. In the first project, a pilot called Butterflies and Spiders in Space, 1,876 middle school students tested a protocol for comparing, at near-real time, the behaviors of orb-weaver spiders and painted lady butterflies living in microgravity (aboard ISS) to those of comparable subjects in students’ classrooms. Teachers reported that, as a result of project activities, 33% of their students designed additional experiments and 80% of students expressed interest in science careers. The second program, Butterflies in Space, enabled students to observe and investigate the life cycle and behaviors of painted lady butterflies living on ISS, and compare them to butterflies being studied in their own classes. Combining this near real-time experiment with hands-on explorations and web-based instructional strategies, Butterflies in Space reached more than 3,000 teachers, representing an estimated 180,000 students (grades 3-6) or more worldwide. It also received international coverage from a variety of media. Investigators at BioServe Space Technologies of the University of Colorado designed and built the chambers in which the spiders and butterflies were housed on ISS, and led technical and logistical operations for both programs. Baylor College of Medicine (BCM) educators and scientists developed the education framework and managed the web-based distribution of project data and teaching resources.
Tomatosphere – To Mars and Beyond: An educational outreach project for primary and secondary schools
The concept of Tomatosphere originated in 1999. Phase one saw 200 000 tomato seeds go into space on November 30, 2000, with Canadian astronaut, Dr. Marc Garneau on STS 97. The seeds were part of a Canada wide experiment for school age children in grades 3 – 6 designed to test the effects of short- term space travel on seed germination. In 2002, the scope of the project expanded to include students in grades 8 – 10. This allows for the inclusion of several objectives related to the science curriculum, including: the International Space Station (ISS), the overall concept of humans in space, and the application of knowledge from space programs to the well-being of humankind on Earth. In 2003, each classroom received three sets of seeds; (a) control group; (b) exposure to a simulated Mars environment; and (c) exposure to a simulated Mars greenhouse environment (reduced atmospheric pressure). The results of this “blind test” submitted by teachers indicated little difference in the germination rates of the three groups of seeds. The next phase scheduled for March 2004 involved an extension of the treatment duration. Also, in 2004, 400 000 tomato seeds were taken to the ISS on a Russian Progress flight. These seeds received the “full treatment” in terms of the environmental conditions in space, reduced gravity and cosmic radiation. While these seeds were in space for 19 months, another group of seeds were used for the experiment – seeds that hibernated at the Arthur C. Clarke Mars Greenhouse on Devon Island in the Canadian High Arctic. In 2005, students dealt with these seeds and those that had been exposed to a simulated space environment, reflecting a replicated breach in the storage system of a vehicle en route to Mars. For the first time, results indicated statistically significant differences among the seed treatments. In 2006, more than 7000 classrooms received the seeds from the ISS (returned to Earth on board Discovery in August, 2005), and a control group. The results of this phase of the project will be available in the late summer of 2006. Changes in germination rates will provide data regarding the availability of an adequate – and required – food supply for future astronauts on a Mars transit journey. This program brings critical questions about the fate of plant seeds in space for extended periods into Canadian and US classrooms and provides students and researchers alike with valuable information relevant to studying the role of plants in space for life support.
Microbial detection and monitoring in advanced life support systems like the International Space Station
Potentially pathogenic microbes and so-called technophiles may form a serious threat in advanced life support systems, such as the International Space Station (ISS). They not only pose a threat to the health of the crew, but also to the technical equipment and materials of the space station. The development of fast and easy to use molecular detection and quantification methods for application in manned spacecraft is therefore desirable and may also be valuable for applications on Earth. In this paper we present the preliminary results of the SAMPLE experiment in which we performed molecular microbial analysis on environmental samples of the ISS as part of an ESA-MAP project.
Recently we demonstrated that the effectiveness of RNAi interference (RNAi) for inhibiting gene expression is maintained during spaceflight in the worm Caenorhabditis elegans and argued for the biomedical importance of this finding. We also successfully utilized green fluorescent protein (GFP)-tagged proteins to monitor changes in GPF localization during flight. Here we discuss potential applications of RNAi and GFP in spaceflight studies and the ramifications of these experiments for the future of space life-sciences research.
Human exploration of outer space will eventually take place. In preparation for this endeavour, it is important to establish the nature of the biological response to a prolonged exposure to the space environment. In one of the recent Soyuz Missions to serve the International Space Station (ISS), the Spanish Soyuz mission in October 2003, we exposed four groups of Drosophila male imagoes to microgravity during the almost eleven days of the Cervantes mission to study their motility behaviour. The groups were three of young flies and one of mature flies, In previous space experiments, we have shown that when imagoes are exposed to microgravity they markedly change their behaviour by increasing their motility, especially if subjected to these conditions immediately after hatching. The constraints of the current Soyuz flights made it impossible to study the early posthatching period. A low temperature cold transport was incorporated as a possible way out of this constraint. It turned out that on top of the space flight effects, the cold treatment by itself, modifies the motility behaviour of the flies. Although the four groups increased their motility, the young flies did it in a much lower extent than the mature flies that had not been exposed to the low temperature during transportation. Nevertheless, the flies flown in the ISS are still more active than the parallel ground controls. As a consequence of the lower motility stimulation in this experiment, a likely consequence of the cold transport step, no effects on the life spans of the flown flies were detected. Together with previous results, this study confirms that high levels of motility behaviour are necessary to produce significant decreases in fly longevity.