Detection of DNA Damage by Space Radiation in Human Fibroblast Cells Flown on the International Space Station

Although charged particles in space have been detected with radiation detectors on board spacecraft since the early discovery of the Van Allen Belt, reports on effects of direct exposure to space radiation in biological systems have been limited. Measurement of biological effects of space radiation has been difficult due to the low dose and low dose rate nature of the radiation environment, and the difficulty in separating the radiation effects from microgravity and other space environmental factors. In astronauts, only a small number of changes, such as increased chromosome aberrations in lymphocytes and early onset of cataracts, attributed primarily to the exposure to space radiation. In a recent experiment, human fibroblast cells were flown on the International Space Station (ISS). Cells fixed on Days 3 and 14 after reaching orbit were analyzed for phosphorylation of a histone protein H2AX by immunofluorescent staining of cells, which is a widely used marker for DNA double strand breaks. The 3-dimensional gamma-H2AX foci were captured with a laser confocal microscope. Quantitative analysis revealed a small fraction of foci that were larger and displayed a track pattern in the flight samples in comparison to the ground control. Human fibroblast cells were also exposed to low dose rate gamma rays, as well as to protons and Fe ions. Comparison of the pattern and distribution of the foci after gamma ray and charged particle exposure to our flight results confirmed that the foci found in the flown cells were indeed induced by space radiation

Corrigendum: Collective search by ants in microgravity

The problem of collective search is a tradeoff between searching thoroughly and covering as much area as possible. This tradeoff depends on the density of searchers. Solutions to the problem of collective search are currently of much interest in robotics and in the study of distributed algorithms, for example to design ways that without central control robots can use local information to perform search and rescue operations. Ant colonies operate without central control. Because they can perceive only local, mostly chemical and tactile cues, they must search collectively to find resources and to monitor the colony's environment. Examining how ants in diverse environments solve the problem of collective search can elucidate how evolution has led to diverse forms of collective behavior. An experiment on the International Space Station in January 2014 examined how ants (Tetramorium caespitum) perform collective search in microgravity. In the ISS experiment, the ants explored a small arena in which a barrier was lowered to increase the area and thus lower ant density. In microgravity, relative to ground controls, ants explored the area less thoroughly and took more convoluted paths. It appears that the difficulty of holding on to the surface interfered with the ants’ ability to search collectively. Ants frequently lost contact with the surface, but showed a remarkable ability to regain contact with the surface

Spiders in space - orb-web-related behaviour in zero gravity

Gravity is very important for many organisms, including web-building spiders. Probably the best approach to study the relevance of gravity on organisms is to bring them to the International Space Station. Here, we describe the results of such an experiment where two juvenile Trichonephila clavipes (L.) (Araneae, Nephilidae) spiders were observed over a 2-month period in zero gravity and two control spiders under otherwise identical conditions on Earth. During that time, the spiders and their webs were photographed every 5 min. Under natural conditions, Trichonephila spiders build asymmetric webs with the hub near the upper edge of the web, and they always orient themselves downwards when sitting on the hub whilst waiting for prey. As these asymmetries are considered to be linked to gravity, we expected the spiders experiencing no gravity to build symmetric webs and to show a random orientation when sitting on the hub. We found that most, but not all, webs built in zero gravity were indeed quite symmetric. Closer analysis revealed that webs built when the lights were on were more asymmetric (with the hub near the lights) than webs built when the lights were off. In addition, spiders showed a random orientation when the lights were off but faced away from the lights when they were on. We conclude that in the absence of gravity, the direction of light can serve as an orientation guide for spiders during web building and when waiting for prey on the hub

International Space Station conditions alter genomics, proteomics, and metabolomics in Aspergillus nidulans.

The first global genomic, proteomic, and secondary metabolomic characterization of the filamentous fungus Aspergillus nidulans following growth onboard the International Space Station (ISS) is reported. The investigation included the A. nidulans wild-type and three mutant strains, two of which were genetically engineered to enhance secondary metabolite production. Whole genome sequencing revealed that ISS conditions altered the A. nidulans genome in specific regions. In strain CW12001, which features overexpression of the secondary metabolite global regulator laeA, ISS conditions induced the loss of the laeA stop codon. Differential expression of proteins involved in stress response, carbohydrate metabolic processes, and secondary metabolite biosynthesis was also observed. ISS conditions significantly decreased prenyl xanthone production in the wild-type strain and increased asperthecin production in LO1362 and CW12001, which are deficient in a major DNA repair mechanism. These data provide valuable insights into the adaptation mechanism of A. nidulans to spacecraft environments

Effects of Spaceflight on Human Induced Pluripotent Stem Cell-Derived Cardiomyocyte Structure and Function.

With extended stays aboard the International Space Station (ISS) becoming commonplace, there is a need to better understand the effects of microgravity on cardiac function. We utilized human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to study the effects of microgravity on cell-level cardiac function and gene expression. The hiPSC-CMs were cultured aboard the ISS for 5.5 weeks and their gene expression, structure, and functions were compared with ground control hiPSC-CMs. Exposure to microgravity on the ISS caused alterations in hiPSC-CM calcium handling. RNA-sequencing analysis demonstrated that 2,635 genes were differentially expressed among flight, post-flight, and ground control samples, including genes involved in mitochondrial metabolism. This study represents the first use of hiPSC technology to model the effects of spaceflight on human cardiomyocyte structure and function

Preparation for and performance of a Pseudomonas aeruginosa biofilm experiment on board the International Space Station

Biofilms are problematic on Earth due to their ability to both degrade the materials upon which they grow and promote infections. Remarkably, 65% of infections and 80% of chronic diseases on Earth are associated with biofilms. The impact of biofilms is even greater in space, as the crew's lives and mission success depend on nominal operation of mechanical systems which can be interrupted by material damage associated with biofilm growth. Furthermore, the isolated confined environment nature of spaceflight may increase the rates of disease transmission. In the case of the International Space Station (ISS), biofilms are an identified problem on the Environmental Control and Life Support System (ECLSS), namely on the water processor assembly (WPA). In late 2019, the bacterial component of the Space Biofilms experiment launched to ISS to (i) characterize the mass, thickness, morphology, and gene expression of biofilms formed in space compared to matched Earth controls, (ii) interrogate the expression of antimicrobial resistance genes, and (iii) test novel materials as potential biofilm control strategies for future ECLSS components. For this, 288 bacterial samples were prepared prior to the launch of the Northrop Grumman CRS-12 mission from NASA's Wallops Flight Facility. The samples were integrated into the spaceflight hardware, BioServe's Fluid Processing Apparatus (FPA), packed in sets of eight in Group Activation Packs (GAP). Half of these samples were activated and terminated on orbit by NASA astronauts Jessica Meir and Christina Koch, while the remaining half were processed equivalently on Earth. The spaceflight bacterial samples of Space Biofilms returned on board the SpaceX CRS-19 Dragon spacecraft in early 2020. We here describe the test campaign implemented to verify the experiment design and confirm it would enable us to achieve the project's scientific goals. This campaign ended with the Experiment Verification Test (EVT), from which we present example morphology and transcriptomic results. We describe in detail the sample preparation prior to flight, including cleaning and sterilization of the coupons of six materials (SS316, passivated-SS316, lubricant impregnated surface, catheter-grade silicone with and without a microtopography, and cellulose membrane), loading and integration of growth media, bacterial inoculum, and the fixative and preservative to enable experiment termination on orbit. Additionally, we describe the performance of the experiment on board the ISS, including crew activities, use of assets, temperature profile, and experiment timeline; all leading to a successful spaceflight experiment. Hence, this manuscript focuses on the steps implemented to ensure the experiment would be ready for spaceflight, as well as ISS and ground operations, with results presented elsewhere. The processes discussed here may serve as a guideline to teams planning their own gravitational microbiology experiments. This material is based upon work supported by the National Aeronautics and Space Administration under Grant No. 80NSSC17K0036