This study presents the first global transcriptional profiling and phenotypic characterization of the major human opportunistic fungal pathogen Candida albicans grown in spaceflight conditions. Microarray analysis revealed that C. albicans subjected to short-term spaceflight culture differentially regulated 454 genes compared to synchronous ground controls which represented 8.4% of the analyzed ORFs. Spaceflight-cultured C. albicans induced genes involved in cell aggregation (similar to flocculation) which was validated by microscopic and flow cytometry analysis. We also observed enhanced random budding of spaceflight-cultured cells as opposed to more normal bipolar budding patterns for ground samples in accordance with the gene expression data. Furthermore genes involved in antifungal agent and stress resistance were differentially regulated in spaceflight including induction of ABC transporters and members of the major facilitator family downregulation of ergosterol-encoding genes and upregulation of genes involved in oxidative stress resistance. Finally downregulation of genes involved in the actin cytoskeleton was observed. Interestingly the transcriptional regulator Cap1 and over 30% of the Cap1 regulon was differentially expressed in spaceflight-cultured C. albicans. A potential role for Cap1 in the spaceflight response of C. albicans is suggested as this regulator is involved in random budding cell aggregation actin cytoskeleton and oxidative stress resistance; all related to observed spaceflight-associated changes of C. albicans. While culture of C. albicans in microgravity potentiates a global change in gene expression that could induce a virulence-related phenotype no increased virulence in a murine intraperitoneal (i.p.) infection model was observed. This study represents an important basis for the assessment of the risk that commensal flora could play during spaceflight missions. Furthermore since the low fluid-shear environment of microgravity is relevant to physical forces encountered by pathogens during the infection process insights gained from this study could identify novel infectious disease mechanisms with downstream benefits for the general public. Cells were grown for 24 hours on the space shuttle or as ground-based controls preserved in RNALater and stored at -80C. Four samples of each flight- and ground-based controls were harvested for microarray analysis. GAP is Group Activation Pack and each GAP contains 8 FPAs. The numbers represent the # assigned to the particular GAP and the number assigned to the specific FPA (1-8) within the indicated GAP. The same hardware is used for the flight samples and the ground samples.

Microgravity alters the immune response to in vitro LPS assault engineered in spaceflight: A multi-omics study Microgravity can facilitate creation of a potent environment for opportunistic infection by augmenting virulence and suppressing the host defense. Presumably extraterrestrial infection may trigger potentially novel bionetworks different from the terrestrial equivalent which could only be probed by investigating the host-pathogen relationship with minimum terrestrial bias. Towards this objective we strategically engineered a cell culture module equipped with a feedback controlled semi-automated platform to expose human endothelial cells to lipopolysaccharide (LPS). The assay was carried out in the STS-135 space shuttle and a concurrent ground study constituted the baseline. Transcriptomic investigation revealed an immune blunting in microgravity; Lbp MyD88 and MD-2 failed to encode proteins responsible for early LPS uptake. Longer exposure results implied that there was a delayed response potentially ineffectual in preventing pathogens from opportunistically modulating the infection network. Lack of recruitment of growth factors and a debilitated apoptosome supported this potential explanation. Certain cytokines such as IL-6 and IL-8 surged in response to LPS insult in microgravity. Contrasting expressions of B2M TIMP-1 and VEGRs suggested impaired pro-survival adaptation and healing mechanisms. The susceptibility of oxidative stress and immune regulation to microgravity compelled further investigation of the respective microRNA modulators such as miR-200a and miR-146b. These miRNAs were expressed differently in response to LPS assaults in different gravitational limits. In conclusion despite a serious drawback attributed to the small sample size we delineated some of the important aspects of the extraterrestrial etiology; more comprehensive follow up studies are warranted. Present study though compromised by the small sample size was able to shade lights on several aspects of immunological responses to the endotoxic assault mediated by uG. Implementing the host-pathogen interactions in the spaceflight and subsequently lysing the cells onboard presented the critical distinguishing features of the present study from the past reports. We identified the CCM of Tissue Genesis Inc. HI as the suitable hardware system to carry out the experiment in the spaceflight. CCM is an automated feedback controlled module that can concurrently support 24 bioreactors following protocols exclusively programmed for individual bioreactor. For this experiment we use samples EA41 EA 47 EA45 and EA155 that were exposed to LPS for 4 hours. Samples EA123 EA165 EA127 EA126 were exposed to LPS for 8Hrs. Samples EA33 EA 125 EA79 and EA 39 were controls in this experiment.

Bacterial behavior has been observed to change during spaceflight. Higher final cell counts enhanced biofilm formation increased virulence and reduced susceptibility to antibiotics have been reported to occur for cells cultured in space . Most of these phenomena are theorized as being an indirect effect of an altered extracellular environment where the carbon source uptake is inhibited and excreted acidic byproducts buildup around the cell due to the lack of gravity-driven transport forces. However to date neither spaceflight results ground-based studies physical measurement techniques nor computational approaches have provided sufficient evidence needed to confirm this model. Gene expression data from the Antibiotic Effectiveness in Space (AES-1) experiment however have now allowed us to look into the biomolecular processes behind these observations and showed a systematic activation of glucose starvation and acid resistance genes. These results corroborate the reduced mass transport model proposed to govern bacterial responses to spaceflight. Furthermore the gene expression data suggests that metabolism was stimulated in space which could play a role in causing the observed increase in bacterial cell concentrations in microgravity. Similarly the decrease in extracellular pH may also be involved with the reported increase in virulence in space.

During space flight bone mineral density is decreased by the influence of osteoclast activation which molecular mechanism is expectantly investigated. In the study of medaka bone development we investigated the system of vertebra formation and firstly identified the presence of osteoclasts in medaka. Moreover osteoclast rsorbing activity was affected by hypergravity indicating the possibility that we can investigate the effect of microgravity on osteoclasts in space. To find this effect we examine the alteration of osteoclast activity under microgravity with the histological analysis or the expression analysis by RNA in-situ hybridization. Furthermore since we have succeeded the establishment of the medaka osteoclast-specific transgenic lines we perform the in-vivo imaging analyses for gene expression and cell mobility. Finally to examine the gravity sensing system we employ tooth and bone as the high density organs which are highly sensitive to gravity and perform the histological analysis and the gene expression analysis of such gravity-sensitive tissues at surrounding pharyngeal teeth and supporting bone.

We investigated differentially regulated and stably expressed genes in human Jurkat T lymphocytic cells in 5min simulated microgravity and hypergravity and compared expression profiles to identify gravity-regulated and unaffected genes as well as adaptation processes. This dataset is part of a series of three and the other two datasets are deposited in GLDS-172 and GLDS-188.

We report how high and low linear energy transfer (LET) radiation microgravity and elevated dietary iron affect colon microbiota (determined by 16S rDNA pyrosequencing) and colon function. Three independent experiments were conducted: 1) fractionated low LET gamma radiation (137Cs 3 Gy RAD) high Fe diet (IRON) (650 mg/kg diet) and a combination of low LET gamma radiation and high Fe diet (IRON+RAD) in male Sprague-Dawley rats; 2) high LET 38Si particle exposure (0.050 Gy) 1/6 G partial weight bearing (PWB) and a combination of high LET38Si particle exposure and PWB in female BalbC/ByJ mice; and 3) 13 d spaceflight in female C57BL/6 mice. For each experiment the colon was resected and feces removed for microbial sequencing analysis on a Roche 454 Genome Sequencer FLX Titanium instrument (Microbiome Core Facility Chapel Hill NC) using the GS FLX Titanium XLR70 sequencing reagents and protocols. Analysis of amplicon sequencing data was carried out using the QIIME pipeline. Low LET radiation high iron diet and spaceflight increased Bacteroidetes and decreased Firmicutes. Low LET radiation high Fe diet and spaceflight did not significantly affect diversity or richness or elevate pathogenic genera. Spaceflight increased Clostridiales and decreased Lactobacillales and similar trends were observed in the experiment using a ground-based model of microgravity suggesting altered gravity may affect colonic microbiota. Microbiota characteriztion in these models is a first step in understanding the impact of the space environment on intestinal health.

We investigated differentially regulated genes in human Jurkat T lymphocytic cells in 20s and 5min microgravity and in hypergravity and compared expression profiles to identify potential gravity-regulated genes and adaptation processes. This dataset is part of a series of three and the other two datasets are deposited in GLDS-188 and GLDS-189.

Microgravity effect on C. elegans gene expression was analysed by whole genome microarray. The worms were cultivated under microgravity for 4 days in the Japanese Module of the International Space Station. C. elegans N2 was exposed microgravity for 4 days. The worms synchronously were cultivated from L1 larvae to adult. There are two control groups onboard 1G and ground 1G control.

The effect of microgravity on gene expression in C.elegans was comprehensively analysed by DNA microarray (Agilent). Overall design: Mix staged C. elegans N2 was exposed microgravity for 10 days.

In an on-going Microbial Observatory experimental investigation on the International Space Station (ISS) multiple bacterial isolates of Biosafety Level 2 (BSL-2) were isolated and identified. The antibiotic susceptibility pattern was tested in these BSL-2 isolates for the following antibiotics: cefazolin ciprofloxacin cefoxitin erythromycin gentamycin oxacillin penicillin rifampin tobramycin and many of the BSL-2 isolates showed multiple drug resistance. Among these isolates 21 strains were chosen for whole genome sequencing (WGS) for a possible lead to develop appropriate countermeasures. In addition the genomic data would enable to determine the influence of microgravity on the pathogenicity and virulence in the BSL-2 microorganisms.