Research Containing: Space Flight

Spaceflight enhances cell aggregation and random budding in Candida albicans

by cfynanon 9 June 2015in Biology & Biotechnology No comment

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 452 genes compared to synchronous ground controls, which represented 8.3% 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 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 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, 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 under the conditions of this study. Collectively, our data represent an important basis for the assessment of the risk that commensal flora could play during human 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.

Response of Pseudomonas aeruginosa PAO1 to low shear modelled microgravity involves AlgU regulation

by cfynanon 9 June 2015in Biology & Biotechnology No comment

As a ubiquitous environmental organism that is occasionally part of the human flora, Pseudomonas aeruginosa could pose a health hazard for the immunocompromised astronauts during long-term missions. Therefore, insights into the behaviour of P. aeruginosa under spaceflight conditions were gained using two spaceflight-analogue culture systems: the rotating wall vessel (RWV) and the random position machine (RPM). Microarray analysis of P. aeruginosa PAO1 grown in the low shear modelled microgravity (LSMMG) environment of the RWV, compared with the normal gravity control (NG), revealed an apparent regulatory role for the alternative sigma factor AlgU (RpoE-like). Accordingly, P. aeruginosa cultured in LSMMG exhibited increased alginate production and upregulation of AlgU-controlled transcripts, including those encoding stress-related proteins. The LSMMG increased heat and oxidative stress resistance and caused a decrease in the oxygen transfer rate of the culture. This study also showed the involvement of the RNA-binding protein Hfq in the LSMMG response, consistent with its previously identified role in the Salmonella LSMMG and spaceflight response. The global transcriptional response of P. aeruginosa grown in the RPM was highly similar to that in NG. Fluid mixing was assessed in both systems and is believed to be a pivotal factor contributing to transcriptional differences between RWV- and RPM-grown P. aeruginosa. This study represents the first step towards the identification of virulence mechanisms of P. aeruginosa activated in response to spaceflight-analogue conditions, and could direct future research regarding the risk assessment and prevention of Pseudomonas infections during spaceflight and in immunocompromised patients.

Transcriptional and proteomic responses of Pseudomonas aeruginosa PAO1 to spaceflight conditions involve Hfq regulation and reveal a role for oxygen

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Assessing bacterial behavior in microgravity is important for risk assessment and prevention of infectious diseases during spaceflight missions. Furthermore, this research field allows the unveiling of novel connections between low-fluid-shear regions encountered by pathogens during their natural infection process and bacterial virulence. This study is the first to characterize the spaceflight-induced global transcriptional and proteomic responses of Pseudomonas aeruginosa, an opportunistic pathogen that is present in the space habitat. P. aeruginosa responded to spaceflight conditions through differential regulation of 167 genes and 28 proteins, with Hfq as a global transcriptional regulator. Since Hfq was also differentially regulated in spaceflight-grown Salmonella enterica serovar Typhimurium, Hfq represents the first spaceflight-induced regulator acting across bacterial species. The major P. aeruginosa virulence-related genes induced in spaceflight were the lecA and lecB lectin genes and the gene for rhamnosyltransferase (rhlA), which is involved in rhamnolipid production. The transcriptional response of spaceflight-grown P. aeruginosa was compared with our previous data for this organism grown in microgravity analogue conditions using the rotating wall vessel (RWV) bioreactor. Interesting similarities were observed, including, among others, similarities with regard to Hfq regulation and oxygen metabolism. While RWV-grown P. aeruginosa mainly induced genes involved in microaerophilic metabolism, P. aeruginosa cultured in spaceflight presumably adopted an anaerobic mode of growth, in which denitrification was most prominent. Whether the observed changes in pathogenesis-related gene expression in response to spaceflight culture could lead to an alteration of virulence in P. aeruginosa remains to be determined and will be important for infectious disease risk assessment and prevention, both during spaceflight missions and for the general public.

Immune system changes during simulated planetary exploration on Devon Island, high arctic

by cfynanon 9 June 2015in Biology & Biotechnology No comment

BACKGROUND: Dysregulation of the immune system has been shown to occur during spaceflight, although the detailed nature of the phenomenon and the clinical risks for exploration class missions have yet to be established. Also, the growing clinical significance of immune system evaluation combined with epidemic infectious disease rates in third world countries provides a strong rationale for the development of field-compatible clinical immunology techniques and equipment. In July 2002 NASA performed a comprehensive immune assessment on field team members participating in the Haughton-Mars Project (HMP) on Devon Island in the high Canadian Arctic. The purpose of the study was to evaluate the effect of mission-associated stressors on the human immune system. To perform the study, the development of techniques for processing immune samples in remote field locations was required. Ten HMP-2002 participants volunteered for the study. A field protocol was developed at NASA-JSC for performing sample collection, blood staining/processing for immunophenotype analysis, whole-blood mitogenic culture for functional assessments and cell-sample preservation on-location at Devon Island. Specific assays included peripheral leukocyte distribution; constitutively activated T cells, intracellular cytokine profiles, plasma cortisol and EBV viral antibody levels. Study timepoints were 30 days prior to mission start, mid-mission and 60 days after mission completion. RESULTS: The protocol developed for immune sample processing in remote field locations functioned properly. Samples were processed on Devon Island, and stabilized for subsequent analysis at the Johnson Space Center in Houston. The data indicated that some phenotype, immune function and stress hormone changes occurred in the HMP field participants that were largely distinct from pre-mission baseline and post-mission recovery data. These immune changes appear similar to those observed in astronauts following spaceflight. CONCLUSION: The immune system changes described during the HMP field deployment validate the use of the HMP as a ground-based spaceflight/planetary exploration analog for some aspects of human physiology. The sample processing protocol developed for this study may have applications for immune studies in remote terrestrial field locations. Elements of this protocol could possibly be adapted for future in-flight immunology studies conducted during space missions.

Immune system dysregulation occurs during short duration spaceflight on board the space shuttle

by cfynanon 9 June 2015in Biology & Biotechnology No comment

BACKGROUND: Post-flight data suggests immunity is dysregulated immediately following spaceflight, however this data may be influenced by the stress effects of high-G entry and readaptation to terrestrial gravity. It is unknown if immunity is altered during spaceflight. METHODS: Blood samples were collected from 19 US Astronauts onboard the Space Shuttle ~24 h prior to landing and returned for terrestrial analysis. Assays consisted of leukocyte distribution, T cell blastogenesis and cytokine production profiles. RESULTS: Most bulk leukocyte subsets (WBC, differential, lymphocyte subsets) were unaltered during spaceflight, but were altered following landing. CD8+ T cell subsets, including cytotoxic, central memory and senescent were altered during spaceflight. T cell early blastogenesis varied by culture mitogen. Functional responses to staphylococcal enterotoxin were reduced during and following spaceflight, whereas response to anti-CD3/28 antibodies was elevated post-flight. The level of virus specific T cells were generally unaltered, however virus specific T cell function was depressed both during and following flight. Plasma levels of IFNalpha, IFNgamma, IL-1beta, IL-4, IL-10, IL-12, and TNFalpha were significantly elevated in-flight, while IL-6 was significantly elevated at R + 0. Cytokine production profiles following mitogenic stimulation were significantly altered both during, and following spaceflight. Specifically, production of IFNgamma, IL-17 and IL-10 were reduced, but production of TNFalpha and IL-8 were elevated during spaceflight. CONCLUSIONS: This study indicates that specific parameters among leukocyte distribution, T cell function and cytokine production profiles are altered during flight. These findings distinguish in-flight dysregulation from stress-related alterations observed immediately following landing.

Monocyte phenotype and cytokine production profiles are dysregulated by short-duration spaceflight

by cfynanon 9 June 2015in Biology & Biotechnology No comment

INTRODUCTION: Immune system dysregulation has been demonstrated to occur during and immediately following spaceflight. As the initial bias and magnitude for an immune response is heavily influenced by monocyte/macrophage secreted cytokines, this study investigated monocyte phenotype and cytokine production patterns following short-duration spaceflight. METHODS: Secreted cytokine profiles were examined by cytometric bead array analysis of culture supernatants following whole blood culture activation with LPS or PMA+ionomycin. Nine short-duration Space Shuttle crewmembers participated in this study. RESULTS: Peripheral monocyte percentages were unaltered postflight. Constitutive monocyte expression of both CD62L and HLA-DR was reduced following spaceflight in a mission-specific fashion. Loss of either molecule indicates a functional disability of monocytes, either by inhibition of adhesion and tissue migration (CD62L) or by impaired antigen presentation (HLA-DR). Following LPS stimulation of monocytes, postflight expression of IL-6, TNFalpha, and IL-10 were significantly reduced (by 43%, 44%, and 41%, respectively) and expression of IL-1b was elevated (65%). IL-8 production was either elevated or reduced in a mission-specific fashion. Following PMA+ionomycin stimulation of all leukocyte populations, only expression of IL-6 was significantly reduced postflight. DISCUSSION: These data indicate that changes in monocyte constitutive phenotype and inflammatory cytokine production occur following short-duration spaceflight, which may impact overall crewmember immunocompetence. Also, monocyte/macrophage function may be highly sensitive to mission specific parameters.

Immune status, latent viral reactivation, and stress during long-duration head-down bed rest

by cfynanon 9 June 2015in Biology & Biotechnology No comment

INTRODUCTION: As logistical access for space research becomes more limited and NASA prepares for exploration-class missions, ground-based spaceflight analogs will increase in importance for biomedical countermeasures development. A monitoring of immune parameters was performed during the NASA Flight Analogs Project bed rest study (without countermeasure); to establish 'control' data against which future studies (with countermeasure) will be evaluated. Some of the countermeasures planned to be evaluated in future studies may impact immune function. METHODS: The immune assessment consisted of: leukocyte subset distribution, early T cell activation, intracellular cytokine profiles, latent viral reactivation, virus specific T cell levels and function, stress hormone levels, and a behavioral assessment using stress questionnaires. RESULTS: In general, subjects did not display altered peripheral leukocyte subsets, constitutive immune activation, altered T cell function, or significant latent viral reactivation (EBV, VZV). Levels of constitutively activated T cells (CD8+/CD69+) and virus-specific T cells (CMV and EBV) decreased during the study. Cortisol levels (plasma and saliva) did not vary significantly during 90-d bed rest. CONCLUSIONS: These data demonstrate the absence of significant immune system alteration and physiological stress during 90-d bed rest, and establish control data against which future studies (including countermeasures) may be compared.

Geometric illusions in astronauts during long-duration spaceflight

by cfynanon 9 June 2015in Biology & Biotechnology No comment

In our previous studies, we have shown that the occurrence of geometric illusions was reduced in vestibular patients who presented signs of otolith disorders and when healthy observers were tilted relative to gravity. We hypothesized that the alteration in the gravitational (otolith) input was responsible for this change, presumably because of a connection between vestibular and visual-spatial cognitive functions. In this study, we repeated similar experiments in astronauts during long-duration spaceflight. In agreement with the data of otolithic patients, the inverted-T geometric illusion was less present in the astronauts in 0 g than in 1g. In addition, the vertical length of drawings made by astronauts in orbit was shorter than that on the ground. This result is also comparable with the otolithic patients who perceived the vertical length of line drawings to be smaller than healthy individuals. We conclude that the impairment in the processing of gravitational input in long-duration astronauts affects their mental representation of the vertical dimension similar to the otolithic patients. The astronauts, however, recover to baseline levels within 1 week after returning to Earth.

Eye movements and motion perception during off-vertical axis rotation after spaceflight

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Constant velocity off-vertical axis rotation (OVAR) provides dynamic linear acceleration stimuli that can be used to assess otolith function. Eight astronauts were rotated in darkness about their longitudinal axis 20 degrees off vertical at low (0.125 Hz) and high (0.5 Hz) frequencies and their responses were compared before and after spaceflight. Eye movements were recorded using infrared videography and perceived motion was evaluated using a joystick with four degrees of freedom – pitch and roll tilt, front-back and lateral translation. Low-frequency OVAR generates tilt otolith-induced responses – modulation of ocular counter-roll and counter-pitch with perceived conical motion path – whereas high-frequency OVAR generates translational otolith-induced responses – modulation of horizontal and vergence slow phase velocity with perceived cylindrical motion path. While there were transient changes in the amplitude of the translational ocular responses on landing day, there were no major changes in the tilt ocular reflexes after adaptation to weightlessness. However, there was an increase in sensitivity to motion perception after spaceflight. Direct comparisons of pre- and postflight stimuli suggested that OVAR on landing day was less provocative of motion sickness than before spaceflight. These results confirm that some otolith reflexes elicited during passive motion may not be altered by short-duration spaceflight – or may readapt very quickly – and that the resolution of sensory conflict associated with postflight recovery involves higher-order neural processes.

Motion perception during tilt and translation after space flight

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Preliminary results of an ongoing study examining the effects of space flight on astronauts' motion perception induced by independent tilt and translation motions are presented. This experiment used a sled and a variable radius centrifuge that translated the subjects forward-backward or laterally, and simultaneously tilted them in pitch or roll, respectively. Tests were performed on the ground prior to and immediately after landing. The astronauts were asked to report about their perceived motion in response to different combinations of body tilt and translation in darkness. Their ability to manually control their own orientation was also evaluated using a joystick with which they nulled out the perceived tilt while the sled and centrifuge were in motion. Preliminary results confirm that the magnitude of perceived tilt increased during static tilt in roll after space flight. A deterioration in the crewmember to control tilt using non-visual inertial cues was also observed post-flight. However, the use of a tactile prosthesis indicating the direction of down on the subject's trunk improved manual control performance both before and after space flight.

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Research Containing: Space Flight

Spaceflight enhances cell aggregation and random budding in Candida albicans

Response of Pseudomonas aeruginosa PAO1 to low shear modelled microgravity involves AlgU regulation

Transcriptional and proteomic responses of Pseudomonas aeruginosa PAO1 to spaceflight conditions involve Hfq regulation and reveal a role for oxygen

Immune system changes during simulated planetary exploration on Devon Island, high arctic

Immune system dysregulation occurs during short duration spaceflight on board the space shuttle

Monocyte phenotype and cytokine production profiles are dysregulated by short-duration spaceflight

Immune status, latent viral reactivation, and stress during long-duration head-down bed rest

Geometric illusions in astronauts during long-duration spaceflight

Eye movements and motion perception during off-vertical axis rotation after spaceflight

Motion perception during tilt and translation after space flight