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

The development of Col-0 and WS plants on orbit differs from that on the ground as demonstrated by the comparison of the gene expression profiles between 4 days old to 8 days old plant in the two environments. The Col-0 plants used different genes reflecting different physiological processes than WS plants suggesting the role of the genetic background in the developmental decisions. The 4 days old Col-0 plant in orbit showed deficit in wax and suberin production relatively to the 8 days old plants the difference unregistered on the ground. There was more dramatic difference in the overexpression of the root system development and anatomical structure development genes in 8 days old plants than in 4 days old plant in both genotypes on the ground than in flight implying smaller root developmental gap between 4 days and 8 days old roots in orbit. The WS plant uniquely showed overexpression of the photosynthesis related genes in the 4 days old roots relative to 8 days old root in the spaceflight environment but not on the ground. The seeds germinated in the novel growth environment of ISS implemented different developmental strategies as captured by the genes expression patterns than seeds developing on the ground.

To understand the molecular mechanisms affected by spaceflight it is essential to achieve high quality sample preservation on-orbit for downstream gene expression analysis. However sample preservation protocols must also be compatible with available equipment and crew time. NASA s Rodent Research (RR) missions have used various methods for euthanasia carcass preservation and tissue preservation. This study extends the sample preservation study performed by GeneLab in GLDS-49 which examined conditions used for the RR-1 mission to include conditions used for multiple RR missions and is designed to help determine factors which may confound data analysis. To determine whether these various factors affect changes in gene expression this ground-based study generated gene expression profiles measured by RNAseq from the livers of 20-21 week-old female C57BL/6J mice. Multiple interacting factors were investigated: 1) To understand how euthanasia protocols affect gene expression when mouse carcasses are slow frozen mice were euthanized by either euthasol injection ketamine/xylazine injection or CO2 inhalation and carcasses slow frozen on dry-ice mimicking carcass preservation in the MELFI on the ISS. Carcasses were thawed and RNA extracted from livers; 2) To understand how carcass preservation protocols affect gene expression mice were euthanized with euthasol and carcasses preserved by flash freezing in liquid nitrogen slow freezing on dry ice or immersion in RNAlater following three-way segmentation. Carcasses were thawed and RNA extracted from livers; 3) To understand how tissue preservation protocols affect gene expression mice were euthanized with euthasol and livers dissected and processed immediately or preserved by flash freezing in liquid nitrogen slow freezing on dry ice or immersion in RNAlater. Liver samples that were processed immediately were homogenized in RLT buffer and then either immediately further processed for RNA extraction or were stored for 70 days at -80C post-homogenization in sample RLT buffer prior to RNA extraction.

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).

Lettuce crops grown in the Veggie payload aboard ISS harvested and returned for chemical microbiological and molecular analysis

To understand the molecular mechanisms affected by spaceflight it is essential to achieve high quality sample preservation on-orbit for downstream gene expression analysis. However sample preservation protocols must also be compatible with available equipment and crew time. NASA s Rodent Research (RR) missions have used various methods for euthanasia carcass preservation and tissue preservation. This study extends the sample preservation study performed by GeneLab in GLDS-49 which examined conditions used for the RR-1 mission to include conditions used for multiple RR missions and is designed to help determine factors which may confound data analysis. To determine whether these various factors affect changes in gene expression this ground-based study generated gene expression profiles measured by RNAseq from the quadriceps of 20-21 week-old female C57BL/6J mice. Multiple interacting factors were investigated: 1) To understand how euthanasia protocols affect gene expression when mouse carcasses are slow frozen mice were euthanized by either euthasol injection ketamine/xylazine injection or CO2 inhalation and carcasses slow frozen on dry-ice mimicking carcass preservation in the MELFI on the ISS. Carcasses were thawed and RNA extracted from quadriceps; 2) To understand how carcass preservation protocols affect gene expression mice were euthanized with euthasol and carcasses preserved by flash freezing in liquid nitrogen slow freezing on dry ice or immersion in RNAlater following three-way segmentation. Carcasses were thawed and RNA extracted from quadriceps; 3) To understand how tissue preservation protocols affect gene expression mice were euthanized with euthasol and quadriceps immediately dissected and preserved by flash freezing in liquid nitrogen slow freezing on dry ice or immersion in RNAlater.

While it has been shown that astronauts suffer immune disorders after spaceflight the underlying causes are still poorly understood and there are many variables to consider when investigating the immune system in a complex environment. Additionally there is growing evidence that suggests that not only is the immune system being altered but the pathogens that infect the host are significantly influenced by spaceflight and ground-based spaceflight conditions. In this study we demonstrate that Serratia marcescens (strain Db11) was significantly more lethal to Drosophila melanogaster after growth on the International Space Station than ground-based controls but that the host immune system is not significantly altered amongst known immune genes. High-throughput sequencing of wild-type (w1118) adult hosts infected with either space or ground-reared S. marcescens revealed few changes in gene expression with 11 genes significantly differentially expressed (q-values <0.05) and only one gene related to the immune system. This data supports the main findings of the paper which state that both spaceflight and low-shear modeled microgravity conditions increase the virulence of this pathogen independent of the host immune system. This data which shows that there are no significant immune-related changes to the host when infected with space-grown sample compared to ground-grown sample provides further evidence that there are likely phenotypic changes to the pathogen itself that is causing increased virulence in spaceflight and in low-shear modeled microgravity. RNA was extracted in triplicate from 2 pooled adult (2-3 day old female) Drosophila melanogaster (w1118) per treatment with 4 total treatment groups (no injection control sham injection with PBS ground bacteria-injected and space bacteria-injected) with poly(A)+ RNA libraries. Samples were multiplexed and sequenced 100bp paired-end ready were sequenced on one lane of the Illumina HiSeq-4000.

Previous spaceflight experiments using Bacillus subtilis have reported altered transcriptome profiles during spaceflight compared to matching ground samples. This study tried to replicate those transcriptome changes using High Aspect Ratio Vessels (HARVs). B. subtilis spores were grown in HARVs using the same protocol and growth conditions as the BRIC-23 spaceflight mission. RNA was extracted from the HARV samples and sequenced for differential expression analysis. HARV differentially expressed genes were then compared to the genes identified as differentially expressed in the BRIC-23 mission to access the accuracy of HARVs as a model for spaceflight experiments.

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 to ground control hiPSC-CMs. Exposure to microgravity on the ISS caused alterations in hiPSC-CM calcium handling. RNA-sequencing analysis demonstrated 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 hiPSCs to model the effects of spaceflight on human cardiomyocyte structure and function.

Prolonged residence of mice in spaceflight is a scientifically robust and ethically ratified model of muscle atrophy caused by continued unloading. Under the Rodent Research Program of NASA we assayed the genomic and metabolomics perturbations in the quadriceps of C57BL/6j male mice that lived on the spaceflight (FLT) or at Ground Control (CTR) for approximately four weeks. Wet weight of quadriceps were significantly reduced in FLT mice. Deep next generation sequencing and untargeted mass spectroscopic assay interrogated the gene-metabolite landscape of same tissues. A majority of top ranked differentially suppressed genes in FLT encode proteins from myosin or troponin family suggesting a sarcomere alteration in space. Significantly enriched gene-metabolite networks were found linked to saromeric integrity immune fitness and oxidative stress response; all inhibited in space as per in silico prediction. A significant loss of FLT mitochondrial DNA copy numbers underlined the energy deprivation associated with spaceflight induced stress and this hypothesis was reinforced by the omics analysis that showed inhibited networks related to protein lipid and carbohydrate metabolism and ATP synthesis and hydrolysis. Finally we reported a list of upstream regulators which could be targeted for next generation therapeutic intervention for the betterment of the musculoskeletal system in male mice subjected to chronic disuse.