Space travel presents unlimited opportunities for exploration and discovery but requires a more complete understanding of the immunological consequences of long-term exposure to the conditions of spaceflight. To understand these consequences better and to contribute to design of effective countermeasures we used the Drosophila model to compare innate immune responses to bacteria and fungi in flies that were either raised on earth or in outer space aboard the NASA Space Shuttle Discovery (STS-121). Microarrays were used to characterize changes in gene expression that occur in response to infection by bacteria and fungus in drosophila that were either hatched and raised in outer space (microgravity) or on earth (normal gravity). Whole Oregon R strain drosophila melanogaster fruit flies either raised on earth or in space that were (1) uninfected (2) infected with bacteria (Escherichia coli) or (3) infected with fungus (Beauveria bassiana) were used for RNA extraction and hybridization on Affymetrix microarrays.

As part of an ongoing effort to catalogue microbial communities inhabiting the International Space Station (ISS) crew-associated environmental samples were collected from the Japanese Kibo Russian and US research modules. Initial analysis based on 16S rRNA gene sequencing identified 11 Bacillus isolates (two from Kibo Japanese Experiment Module (JEM) four from US Segment Harmony Node 2 and five from Russian Segment Zvezda module sites) all belonging to the Bacillus anthracis-B. cereus-B. thuringiensis group. Isolates were further characterized by whole genome comparative analysis. Each isolate was sequenced assembled and aligned against all members of the B. cereus sensu lato group. Based on genome size estimates (5.2 - 5.3 Mbp) strain nucleotide identity (>99.99%) and maximum likelihood phylogenetic placement all isolates were found to exhibit a very high level of similarity. With respect to gene content all isolates were inspected for cry proteins common to B. thuringiensis as well as toxins specific to B. cereus. No significant hits were found to any known cry genes while full-length matches were found for multiple B. cereus toxin genes. Finally the isolates were closely screened against B. anthracis genomes and no B. anthracis signatures were identified. Notably all isolates contained the plcR ancestral C allele and lacked significant hits to pXO1 and pXO2 plasmids and toxins. The collective results from the analysis confirm that these strains belong to a previously uncharacterized Bacillus species hereafter referred to as Bacillus issensis. The type strain is ISSFR-003T (=NRRL B-65389T= DSMZ 101676T).

Comparative gene expression in skin between mice maintained in microgravity (0g) and normogravity (1g) environment. Six male C57Bl/J10 mice were housed for 91 days in the specially designed Mouse Drawer System in weightlessness aboard the International Space Station. Three wild-type mice (WT) and three transgenic mice overexpressing the osteogneic factor PTN/OSF1 under the control of the human bone specific ostecalcin promoter (Tg) were used in the experiment. During the 3-month stay on the ISS 3 mice unfortunately died leaving 2 Tg and 1 WT. MDS tissue sharing program allowed several teams to study various tissues from these mice. Our aim was to investigate the effect of such a long period of microgravity on skin physiology by morphological biochemical and genomewide analyses by comparison to similar mice on ground. Gene expression in the skin of 3 space mice and of 3 ground mice was analyzed by microarray. As this unique experiment performed on 3 mice limits the power of statistical analyis as the transgene PTN/OSF1 was not overexpressed in skin and as a pair wise Pearson s correlation rates between the individual levels of expressed transcripts in the WT and the Tg mice were not significantly different from each other in one experimental group (space or ground) data from the 3 mice were combined to compare results from the space an ground groups.

We address a key baseline question of whether gene expression changes are induced by the orbital environment and then we ask whether undifferentiated cells cells presumably lacking the typical gravity response mechanisms perceive spaceflight. Arabidopsis seedlings and undifferentiated cultured Arabidopsis cells were launched in April 2010 as part of the BRIC-16 flight experiment on STS-131. Biologically replicated DNA microarray and averaged RNA digital transcript profiling revealed several hundred genes in seedlings and cell cultures that were significantly affected by launch and spaceflight. The response was moderate in seedlings; only a few genes were induced by more than 7-fold and the overall intrinsic expression level for most differentially expressed genes was low. In contrast cell cultures displayed a more dramatic response with dozens of genes showing this level of differential expression a list comprised primarily of heat shock-related and stress-related genes. This baseline transcriptome profiling of seedlings and cultured cells confirms the fundamental hypothesis that survival of the spaceflight environment requires adaptive changes that are both governed and displayed by alterations in gene expression. The comparison of intact plants with cultures of undifferentiated cells confirms a second hypothesis: undifferentiated cells can detect spaceflight in the absence of specialized tissue or organized developmental structures known to detect gravity.

To understand how humans adapt to space environments many experiments can be conducted on astronauts while they work aboard the Space Shuttle or the International Space Station (ISS). We also need animal experiments that can apply to human models and help prevent or solve the physical issues we face in space travel. The medaka is a suitable model fish for studying space adaptation because in the second International Microgravity Laboratory mission in 1994 adult fish mated successfully in space during 15 days of flight. In 2012 another space experiment Medaka Osteoclast was performed. Male and female fish (6 weeks old at launching) were maintained in the Aquatic Habitat system for 2 months in the ISS. The RNA-seq analysis of tissues from these fish will revealed tissue-specific responsiveness and common stress responses during space adaptation.

We used microarrays to investigate the effects of microgravity and space radiation on the genome-wide expression of the C. elegans. Three technical replicates of wild type C. elegans (CC1 strain) which exposed to space radiation are analyzed along with ground control.

The effect of microgravity on gene expression in C.elegans was comprehensively analysed by DNA microarray. This is the first DNA microarray analysis for C.elegans grown under microgravity. Hyper gravity and clinorotation experiments were performed as reference against the flight experiment.

Spaceflight results in a number of adaptations to skeletal muscle including atrophy and shifts towards faster muscle fiber types. To identify changes in gene expression that may underlie these adaptations microarray expression analysis was performed on gastrocnemius from mice flown on the STS-108 shuttle flight (11 days 19 hours) versus mice maintained on earth for the same period. Additionally to identify changes that were due to unloading and reloading microarray analyses were conducted on calf muscle from ground-based mice subjected to hindlimb suspension (12 days) and mice subjected to hindlimb suspension plus a brief period of reloading (3.5 hours) to simulate the time between landing and sacrifice of the spaceflight mice.

Microbes interact with humans in complex ways and understanding how they respond to the spaceflight environment is important to the success of future manned spaceflight missions. The BRIC-23 mission was designed to measure the response of Bacillus subtilis and Staphylococcus aureus to the spaceflight environment. This experiment aimed to produce high quality omics data from B. subtilis and S. aureus grown aboard the International Space Station (ISS) to allow comparison to matched ground controls. There were two primary objectives for this experiment: (1) Demonstrate all post-flight processes and operations required for successful completion of GeneLab Reference Missions conducted on ISS and (2) Generate high quality GeneLab Reference Mission omics data sets for two prokaryotic model organisms Bacillus subtilis and Staphylococcus aureus. Freezing Control Experiment: The BRIC hardware has significant thermal inertia thus the freezing rate of samples placed at -80 C is quite slow. This could affect RNA-sequencing proteomic and metabolic data sets. In an effort to understand how slow freezing could affect these data sets a control experiment was designed in which B. subtilis and S. aureus were grown in petri plates and either slow frozen to -80 C at a rate matching the BRIC-23 spaceflight samples or processed immediately to harvest RNA and protein. B.subtilis omics data is deposited in GLDS-138.

Transcriptional crosstalk between mammary gland liver and adipose tissue Experiment Overall Design: Pregnant and Lactating rats exposed to 3 gravity conditions