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.

Harsh environmental conditions including microgravity and radiation during prolonged spaceflights are known to alter hepatic metabolism. Our studies have focused on the analysis of possible changes in metabolic pathways in livers of mice which experienced 30 days of spaceflight with and without an additional re-adaption period of 7 days compared to control mice on Earth. Utilizing shotgun mass spectrometry and label-free quantification we performed proteomic profiling to investigate mice livers from the spaceflight project xe2 x80 x9cBion-M 1 xe2 x80 x9d. No significant alterations in protein levels were observed between control mice liver and spaceflight mice which is possibly caused by insufficient fold change detection combined with high variances within the groups. In contrast our results show that more than a third of the quantified protein levels are altered comparing the liver proteome of mice with and without re-adaption time after their spaceflight. Proteins related to amino acid metabolism showed higher levels after re-adaption which may indicate higher rates of gluconeogenesis. Members of the peroxisome proliferator-activated receptor pathway reconstitute their level after 7 days due to a decrease in fold change which indicates decreased signs of non-alcoholic fatty liver disease. Moreover bile acid secretion regenerates on Earth due to reconstitution of related transmembrane proteins and elevated levels of the drug-metabolising enzymes belonging to the CYP superfamily decrease 7 days after the spaceflight. Thus our study demonstrates reconstitution of pharmacological response and early signs of non-alcoholic fatty liver disease recover within 7 days whereas glucose uptake should be monitored due to alterations in gluconeogenesis.

These data were generated from mice that were skeletally unloaded using antiorthostatic suspension (AOS). Mice were either subjected to AOS or tail restraint without unloading for two weeks before injection with saline +/- tetanus toxoid +/- Cpg. Mice were unloaded for an additional two weeks before being sacrificed. This set of data focused on bone marrow. The spleen data are available in the GLDS-201.

In the Rocket Science citizen science project Eruca sativa (salad rocket) seeds stored for six months on board of the International Space Station caused delayed crop establishment. Here we investigated the physiological and molecular mechanisms underpinning the spaceflight effects on dry seeds. We found that Space seed germination vigor was reduced and aging sensitivity increased but the spaceflight did not compromise seed viability and the development of normal seedlings. Comparative analysis of the transcriptomes (using RNASeq) in dry seeds and upon controlled artificial aging treatment (CAAT) revealed differentially expressed genes (DEGs) associated with spaceflight and ageing. DEG categories enriched by spaceflight and CAAT included transcription and translation with reduced transcript abundances for 40S and 60S ribosomal subunit genes. Among the spaceflight-up DEGs were Heat Shock Protein (HSP) DNAJ-related chaperones a Heat Shock Factor (HSFA7a-like) and components of several DNA repair pathways (e.g. ATM DNA ligase1). The response to radiation category was especially enriched in spaceflight-up DEGs including HSPs catalases and the transcription factor HY5. The major finding from the physiological and transcriptome analysis is that spaceflight causes vigor loss and partial ageing during air-dry seed storage for which space environmental factors and consequences for seed storage during spaceflights are discussed.

Four days old rice calli were selected to grow 324h under spaceflight controls 1g-flight controls and 1g-ground controls. We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during this process. Rice calli were selected at different treatment for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain the diff-genes that caused by the microgravity.

We investigated differentially regulated genes in human myelomonocytic U937 cells in 20s and 5min microgravity and in hypergravity and compared expression profiles to identify potential gravity-regulated genes and adaptation processes. Overall design: Human myelomonocytic U937 cells were exposed to altered gravity during a sounding rocket mission.

The Space Algae Experiment Verification Test completed a competitive growth selection on mutagenized microalgae in a series of batch cultures that were passaged for three growth cycles. The experiment was conducted with ground control conditions similar to those actually used in spaceflight. Cultures were limited in growth rate due to gas permeable membranes to provide oxygen and carbon dioxide exchange and a lack of agitation to mix the cells throughout the liquid media. Two strains were grown and three biological replicates were completed for each strain. Specific variables (factors) tested were: 1) The effect of UVC mutagenesis was tested by sampling the algae cultures prior to mutagenesis. 2) The sensitivity of different strains was tested by conducting the experiment with a wild-type and cell wall mutant (cw15) strain. 3) The effect of competitive growth was tested by sampling each biological replicate experiment at the end of each growth cycle. 4) The effect of live culture storage in the dark was tested by sampling each growth cycle twice. At the time of passage a sample of cells was pelleted and frozen. The cultures were then stored in the dark in a Cargo Transport Bag (CTB) to simulate storage on the ISS and return of cultures. This factor tested whether cultures could be stored alive in the dark or if samples needed to be fixed at the time of passage in order to get an accurate representation of the genetic variation in each cycle of growth. Paired-end whole genome sequencing was completed for 38 samples: 2 strains pre mutagenesis and 2 strains X 3 biological replicates X 3 growth cycles X 2 storage conditions.

The BRIC-21 mission was designed to identify the response of Bacillus subtilis to the human spaceflight environment. For this mission samples were grown in rich-medium using the Biological Research in Canister Petri Dish Fixation Units (BRIC-PDFU) spaceflight hardware. B. subtilis spores were inoculated during spaceflight grown at the ambient ISS temperature and frozen in the onboard -80 C freezer prior to returning to Earth. RNA was extracted from samples grown onboard the International Space Station (ISS) and matching Ground Controls for transcriptome analysis.

Rubinsicoccus jplei genome a novel genus isolated from JPL spacecraft environment where Mars 2020 subsystems are assembled.

The objective of the Rodent Research-9 (RR-9) mission was to use mice to understand the molecular basis of phenomena that affect astronauts during long-duration spaceflight particularly visual impairment and joint tissue degradation. To this end a flight group (FLT) of 10-week-old male C57BL/6J mice was launched from Kennedy Space Center (KSC) on 8/14/2017 and housed in Rodent Habitats on the ISS for 33 days before being returned alive to Earth. After splashdown in the Pacific Ocean the animals were transported to Loma Linda University (LLU) for testing euthanasia and dissection on 9/18/2018. A Basal Control (BSL) was housed in standard cages at Kennedy Space Center (KSC) and euthanized one day after launch of the FLT animals (8/15/2017). Ground Control (GC) and Vivarium Control (VIV) studies were planned to commence at KSC approximately one-week after the conclusion of the flight experiments. However all the GC and VIV mouse studies at KSC had to be cancelled due to Hurricane Irma and potential adverse effects on the animal housing facility. The GC and VIV studies were therefore rescheduled and begun in May 2018. The GC was euthanized and dissected 6/18/2018 - 6/20/2018 while the VIV was euthanized and dissected 6/22/2018 - 6/23/2018. Because this resulted in a different cohort of mice being used for the GC and VIV controls as compared to the flight (FLT) and basal (BSL) groups two cohort controls were included in the study. The first Cohort Control 1 (CC_C1) was from the same cohort as the FLT and BSL animals and was sacrificed and dissected 4 days after the FLT group (9/22/2017). The second Cohort Control 2 (CC_C2) was from the same cohort as the GC and VIV animals and was sacrificed and dissected 2-8 days after the GC and VIV groups (6/24/2018 - 6/26/2018). The CC_C1 and CC_C2 groups were housed in standard cages and fed standard chow in contrast to all other groups which received Rodent Foodbars. To clarify the connections between treatment groups and animal cohorts the following group abbreviations are used in the sample metadata: Flight (FLT_C1); Basal (BSL_C1); Ground Control (GC_C2); Vivarium Control (VIV_C2) Cohort Control 1 (CC_C1); Cohort Control 2 (CC_C2). Fecal pellets were isolated directly from mice during dissection and preserved by flash freezing in liquid nitrogen before stored at -80 C. DNA was then extracted shotgun metagenomic libraries generated and libraries sequenced (target 10 M clusters at PE 250 bp). Metagenomic data was generated from the following groups: Basal Control (n=5) Ground Control (n=5) Vivarium Control (n=5) Cohort Control 1 (n=5) Cohort Control 2 (n=5) Flight (n=5).