Spaceflight Conditions Research Articles

The Effects of Benzofuran-2-Carboxylic Acid Derivatives as Countermeasures in Immune Modulation and Cancer Cell Inhibition

Microgravity and radiation exposure experienced during space flights result in immune system suppression. In long-term spaceflight, the crew is exposed to space radiation, microgravity, infectious agents from other crew members, and microbial contamination, all of which have a significant impact on the body’s immune system and may contribute to the development of autoimmune diseases, allergic reactions, and/or cancer initiation. Many studies have revealed strong effects of microgravity on immune cell function, and microgravity is now considered as one of the major causes of immune dysfunction during space flight (Sundaresan, Int. J. Transp. Phenom. 12(1-2), 93–100, 2011; Martinelli et al., IEEE Eng. Biol. Med. 28(4), 85–90, 2009). We screened two newly synthetized derivatives of benzofuran 2-carboxylic acid, KMEG and KM12. The former KMEG was assessed for lymphoproliferative activities while the latter, KM12, was used in an array of cancer cell lines for testing its cancer inhibiting effects. For ground-based studies, synthetic benzofuran-2-carboxylic acid derivatives were assessed for biological effects in several scenarios, which involved exposure to modeled microgravity and radiation, as well as their immune enhancement and anti-cancer effects. Initial findings indicate that the benzofuran-2-carboxylic acid derivatives possibly have immune enhancing and anti-tumor properties in human lymphocytes and cancer cells exposed to analog spaceflight conditions modeled microgravity and γ-radiation).

A whole-genome microarray study of Arabidopsis thaliana semisolid callus cultures exposed to microgravity and nonmicrogravity related spaceflight conditions for 5 days on board of Shenzhou 8.

The Simbox mission was the first joint space project between Germany and China in November 2011. Eleven-day-old Arabidopsis thaliana wild type semisolid callus cultures were integrated into fully automated plant cultivation containers and exposed to spaceflight conditions within the Simbox hardware on board of the spacecraft Shenzhou 8. The related ground experiment was conducted under similar conditions. The use of an in-flight centrifuge provided a 1 g gravitational field in space. The cells were metabolically quenched after 5 days via RNAlater injection. The impact on the Arabidopsis transcriptome was investigated by means of whole-genome gene expression analysis. The results show a major impact of nonmicrogravity related spaceflight conditions. Genes that were significantly altered in transcript abundance are mainly involved in protein phosphorylation and MAPK cascade-related signaling processes, as well as in the cellular defense and stress responses. In contrast to short-term effects of microgravity (seconds, minutes), this mission identified only minor changes after 5 days of microgravity. These concerned genes coding for proteins involved in the plastid-associated translation machinery, mitochondrial electron transport, and energy production.

Functioning of the kidneys and human body fluids during five-day immersion

Renal function and body composition, including fluids, were studied during space flight conditions imitation with five-day dry immersion in 14 apparently healthy men. Noninvasive research of water spaces of the body was performed using methods of bioimpedance analysis (BIA). It was shown that renal excretion of fluids increased and negative aqueous balance developed during dry immersion. Bioimpedance research revealed decreased volume of the total body and the extracellular fluids, and the amount of circulating plasma also decreased. Thus, under the conditions of immersion, hypohydration developed in the body. In the recovery period, these changes in the hydration level rapidly returned to the initial values. The lean body mass during immersion was reduced insignificantly and the amount of fat, in contrast, exceeded the baseline levels. Furthermore, the experiment showed technical impossibility to obtain reliable impedancemetry data under the dry immersion conditions.

Hind limb unloading, a model of spaceflight conditions, leads to decreased B lymphopoiesis similar to aging.

Within the bone marrow, the endosteal niche plays a crucial role in B-cell differentiation. Because spaceflight is associated with osteoporosis, we investigated whether changes in bone microstructure induced by a ground-based model of spaceflight, hind limb unloading (HU), could affect B lymphopoiesis. To this end, we analyzed both bone parameters and the frequency of early hematopoietic precursors and cells of the B lineage after 3, 6, 13, and 21 d of HU. We found that limb disuse leads to a decrease in both bone microstructure and the frequency of B-cell progenitors in the bone marrow. Although multipotent hematopoietic progenitors were not affected by HU, a decrease in B lymphopoiesis was observed as of the common lymphoid progenitor (CLP) stage with a major block at the progenitor B (pro-B) to precursor B (pre-B) cell transition (5- to 10-fold decrease). The modifications in B lymphopoiesis were similar to those observed in aged mice and, as with aging, decreased B-cell generation in HU mice was associated with reduced expression of B-cell transcription factors, early B-cell factor (EBF) and Pax5, and an alteration in STAT5-mediated IL-7 signaling. These findings demonstrate that mechanical unloading of hind limbs results in a decrease in early B-cell differentiation resembling age-related modifications in B lymphopoiesis.

Device for Noninvasive Determination of Human Body Composition under Spaceflight Conditions

A portable polysegment bioimpedancometer for noninvasive monitoring of human body composition (including fluid media) under spaceflight conditions is described. The device provides real-time monitoring of the dynamics of human body composition under conditions of long-term spaceflight in a space station.

Metabolic Consequences of Skeletal Muscle Atrophy Caused by Space Flight Conditions

The review aims to analyze the relationship between metabolic changes in muscle tissue occurring in space flight conditions and other body systems during the developing atrophy. The data about the activity of various enzyme systems in muscles, liver and myocardium are presented. It was shown that during the transformation of postural muscles, metabolism from the lipid to carbohydrate, the activity of gluconeogenesis in the liver pathways is increased and the intensity of the oxygen consumption by the heart muscle changes.

Biological and metabolic response in STS-135 space-flown mouse skin

There is evidence that space flight condition-induced biological damage is associated with increased oxidative stress and extracellular matrix (ECM) remodeling. To explore possible mechanisms, changes in gene expression profiles implicated in oxidative stress and in ECM remodeling in mouse skin were examined after space flight. The metabolic effects of space flight in skin tissues were also characterized. Space Shuttle Atlantis (STS-135) was launched at the Kennedy Space Center on a 13-day mission. Female C57BL/6 mice were flown in the STS-135 using animal enclosure modules (AEMs). Within 3–5 h after landing, the mice were euthanized and skin samples were harvested for gene array analysis and metabolic biochemical assays. Many genes responsible for regulating production and metabolism of reactive oxygen species (ROS) were significantly (p < 0.05) altered in the flight group, with fold changes >1.5 compared to AEM control. For ECM profile, several genes encoding matrix and metalloproteinases involved in ECM remodeling were significantly up-/down-regulated following space flight. To characterize the metabolic effects of space flight, global biochemical profiles were evaluated. Of 332 named biochemicals, 19 differed significantly (p < 0.05) between space flight skin samples and AEM ground controls, with 12 up-regulated and 7 down-regulated including altered amino acid, carbohydrate metabolism, cell signaling, and transmethylation pathways. Collectively, the data demonstrated that space flight condition leads to a shift in biological and metabolic homeostasis as the consequence of increased regulation in cellular antioxidants, ROS production, and tissue remodeling. This indicates that astronauts may be at increased risk for pathophysiologic damage or carcinogenesis in cutaneous tissue.

Gene Expression Changes Broadly for C. albicans Subjected to Microgravity

Subject to microgravity on space flights, the fungal opportunistic pathogen Candida albicans differentially regulates 452 genes compared to such cells that are subject instead to full gravity on Earth, according to Cheryl Nickerson from the Biodesign Institute at Arizona State University in Tempe and her collaborators from nearly a dozen institutions. This study marks the “first global transcriptional profiling and phenotypic characterization of [this pathogen] under spaceflight conditions,” they note. Details appeared December 2013 in PLOS ONE (doi:10.1371/journal.pone.0080677).

A case for using ground-based thermal inertia measurements to detect Martian caves.

Martian caves are regarded as one of the most interesting locations in which to search for life on the planet. Data obtained during the MARS2013 expedition at Hamar Laghdad Ridge in the Tafilalt region of Morocco indicate that even small cavities can display thermal behavior that is characteristic for caves. For example, temperature in a cavity equaled 14°C±0.1°C before sunrise, which was higher than the temperature of the ambient air (10°C±0.1°C) and proximate rocks (9°C±0.1°C) at the same time. Within 30 min after sunrise, when the temperature of surrounding rocks corresponded to 15°C, this thermal relationship reversed. Measurements were conducted under simulated spaceflight conditions, including near-real-time interpretation of data that were acquired in a complex flight planning environment. We conclude that using ground-based thermal contrast measurements, in 7-14 μm band before and after sunset, is an effective method for Mars astronauts to identify caves, possibly superior to usage of space-based or ground-penetrating data.

Multidisciplinary Russian biomedical research in space

Research activities on a comprehensive multidisciplinary program are vital for enhancement of the system of crew׳s medical care, environmental health and hygiene in space missions. The primary goal of the program must be identification of patterns, intensity and dynamics of structural and functional shifts in organism induced by an aggregate of spaceflight factors including microgravity, isolation, artificial environment, space radiation, etc. Also, the program must pursue differential assessment of emerging deviations from the standpoint of adequacy to the spaceflight conditions and prospects of returning to Earth and guide the development of principles, methods and techniques necessary to maintain health and working capacity of humans during short- and long-duration missions and on return to Earth. Over 50 years, since 1963, the IBMP researchers apply systemic and innovational approaches to fundamental and exploratory studies in the fields of medical sciences, radiation biology, engineering science, biotechnology, etc. with participation of various biological specimens and human volunteers. Investigations aboard manned spacecrafts and biological satellites as well as in ground-based laboratories further enhancement of the medical care system for crews on orbital and remote space missions; they give insight into the fundamental problems of gravitational physiology and biology, psychophysiology, radiation biology, and contribute thereby to the development of knowledge, methods and technologies, as well as medical and scientific equipment.

Three Weeks of Murine Hindlimb Unloading Induces Shifts from B to T and from Th to Tc Splenic Lymphocytes in Absence of Stress and Differentially Reduces Cell-Specific Mitogenic Responses

Extended space missions are known to induce stress and immune dysregulation. Hindlimb unloading is a ground-based model used to reproduce most spaceflight conditions. The aim of this study was to better characterize the consequences of prolonged exposure to hindlimb unloading on murine splenic lymphocyte sub-populations. To ensure that the observed changes were not due to tail restraint but to the antiorthostatic position, three groups of mice were used: control (C), orthostatic restrained (R) and hindlimb unloaded (HU). After 21 days of exposure, no difference in serum corticosterone levels nor in thymus and spleen weights were observed between HU mice and their counterparts, revealing a low state of stress. Interestingly, flow cytometric analyses showed that B cells were drastically reduced in HU mouse spleens by 59% and, while the T cells number did not change, the Th/Tc ratio was decreased. Finally, the use of a fluorescent dye monitoring lymphoproliferation demonstrated that lymphocyte response to mitogen was reduced in Th and Tc populations and to a greater extent in B cells. Thus, we showed for the first time that, even if restraint has its own effects on the animals and their splenic lymphocytes, the prolonged antiorthostatic position leads, despite the absence of stress, to an inversion of the B/T ratio in the spleen. Furthermore, the lymphoproliferative response was impaired with a strong impact on B cells. Altogether, these results suggest that B cells are more affected by hindlimb unloading than T cells which may explain the high susceptibility to pathogens, such as gram-negative bacteria, described in animal models and astronauts.

Comparison of space flight and heavy ion radiation induced genomic/epigenomic mutations in rice (Oryza sativa)

Rice seeds, after space flight and low dose heavy ion radiation treatment were cultured on ground. Leaves of the mature plants were obtained for examination of genomic/epigenomic mutations by using amplified fragment length polymorphism (AFLP) and methylation sensitive amplification polymorphism (MSAP) method, respectively. The mutation sites were identified by fragment recovery and sequencing. The heritability of the mutations was detected in the next generation. Results showed that both space flight and low dose heavy ion radiation can induce significant alterations on rice genome and epigenome (P<0.05). For both genetic and epigenetic assays, while there was no significant difference in mutation rates and their ability to be inherited to the next generation, the site of mutations differed between the space flight and radiation treated groups. More than 50% of the mutation sites were shared by two radiation treated groups, radiated with different LET value and dose, while only about 20% of the mutation sites were shared by space flight group and radiation treated group. Moreover, in space flight group, we found that DNA methylation changes were more prone to occur on CNG sequence than CG sequence. Sequencing results proved that both space flight and heavy ion radiation induced mutations were widely spread on rice genome including coding region and repeated region. Our study described and compared the characters of space flight and low dose heavy ion radiation induced genomic/epigenomic mutations. Our data revealed the mechanisms of application of space environment for mutagenesis and crop breeding. Furthermore, this work implicated that the nature of mutations induced under space flight conditions may involve factors beyond ion radiation.

Comparison of space flight and heavy ion radiation induced genomic/epigenomic mutations in rice (Oryza sativa)

Rice seeds, after space flight and low dose heavy ion radiation treatment were cultured on ground. Leaves of the mature plants were obtained for examination of genomic/epigenomic mutations by using amplified fragment length polymorphism (AFLP) and methylation sensitive amplification polymorphism (MSAP) method, respectively. The mutation sites were identified by fragment recovery and sequencing. The heritability of the mutations were detected in the next generation. Results showed that both space flight and low dose heavy ion radiation can induce significant alterations on rice genome and epigenome (P<0.05). For both genetic and epigenetic assay, while there was no significant difference in mutation rates and their ability to be inherited to the next generation, the site of mutations differed between the space flight and radiation treated groups. More than 50% of the mutation sites were shared by two radiation treated groups, radiated with different LET value and dose, while only about 20% of the mutation sites were shared by space flight group and radiation treated group. Moreover, in space flight group, we found that DNA methylation changes were more prone to occur on CNG sequence than CG sequence. Sequencing results proved that both space flight and heavy ion radiation induced mutations were wide spread on rice genome including coding region and repeated region. Our study described and compared the characters of space flight and low dose heavy ion radiation induced genomic/epigenomic mutations. Our data revealed the mechanisms of application of space environment for mutagenesis and crop breeding. Furthermore, this work implicated that the nature of mutations induced under space flight conditions may involve factors beyond ion radiation.

Transcriptomic and proteomic responses of Serratia marcescens to spaceflight conditions involve large-scale changes in metabolic pathways

The microgravity environment of spaceflight expeditions has been associated with altered microbial responses. This study explores the characterization of Serratia marcescensis grown in a spaceflight environment at the phenotypic, transcriptomic and proteomic levels. From November 1, 2011 to November 17, 2011, a strain of S. marcescensis was sent into space for 398h on the Shenzhou VIII spacecraft, and ground simulation was performed as a control (LCT-SM213). After the flight, two mutant strains (LCT-SM166 and LCT-SM262) were selected for further analysis. Although no changes in the morphology, post-culture growth kinetics, hemolysis or antibiotic sensitivity were observed, the two mutant strains exhibited significant changes in their metabolic profiles after exposure to spaceflight. Enrichment analysis of the transcriptome showed that the differentially expressed genes of the two spaceflight strains and the ground control strain mainly included those involved in metabolism and degradation. The proteome revealed that changes at the protein level were also associated with metabolic functions, such as glycolysis/gluconeogenesis, pyruvate metabolism, arginine and proline metabolism and the degradation of valine, leucine and isoleucine. In summary S. marcescens showed alterations primarily in genes and proteins that were associated with metabolism under spaceflight conditions, which gave us valuable clues for future research.

Genome-wide analysis of microRNA and gene expression profiles in human lymphocytes cultured in modeled microgravity during the DNA-damage response to γ-rays

Perturbations during the DNA-damage response (DDR) originated from alteration of the microRNA-mediated gene regulation can contribute to carcinogenesis initiation. We analyzed the expression profile of microRNAs (miRNAs), which act as post-transcriptional regulators of gene expression, in human peripheral blood lymphocytes (PBLs) incubated in normal gravity (1g) and in modeled microgravity (MMG) during DDR induced by irradiation with γ-rays. We aimed at identifying radio-responsive miRNAs which were affected by MMG incubation during the repair time, altering the cellular response to ionizing radiation (IR). Our results show that miRNA expression profile is dose- and time-dependent in both gravity conditions and that MMG decreased the number of radio-responsive miRNAs. Moreover, we found that several miRNAs such as miR-27a, miR-144, miR-200a, miR-598, miR-650 were differentially expressed due to the combined action of IR and microgravity. Integrated analyses of miRNA and mRNA expression profiles carried out on PBLs derived from the same donors allowed to identify significant miRNA–mRNA anti-correlations of DDR-pathway. The transcripts of ATM and p53 genes were down-regulated in PBLs incubated in MMG during the repair time after IR, together with several anti-correlated genes of p53-pathway. Our findings demonstrate that simulated space flight conditions could influence miRNA expression profile altering the DDR to IR.

Spaceflight Enhances Cell Aggregation and Random Budding in Candida albicans

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.

Differential gene expression profile and altered cytokine secretion of thyroid cancer cells in space

This study focuses on the effects of short-term [22 s, parabolic flight campaign (PFC)] and long-term (10 d, Shenzhou 8 space mission) real microgravity on changes in cytokine secretion and gene expression patterns in poorly differentiated thyroid cancer cells. FTC-133 cells were cultured in space and on a random positioning machine (RPM) for 10 d, to evaluate differences between real and simulated microgravity. Multianalyte profiling was used to evaluate 128 secreted cytokines. Microarray analysis revealed 63 significantly regulated transcripts after 22 s of microgravity during a PFC and 2881 after 10 d on the RPM or in space. Genes in several biological processes, including apoptosis (n=182), cytoskeleton (n=80), adhesion/extracellular matrix (n=98), proliferation (n=184), stress response (n=268), migration (n=63), angiogenesis (n=39), and signal transduction (n=429), were differentially expressed. Genes and proteins involved in the regulation of cancer cell proliferation and metastasis, such as IL6, IL8, IL15, OPN, VEGFA, VEGFD, FGF17, MMP2, MMP3, TIMP1, PRKAA, and PRKACA, were similarly regulated under RPM and spaceflight conditions. The resulting effect was mostly antiproliferative. Gene expression during the PFC was often regulated in the opposite direction. In summary, microgravity is an invaluable tool for exploring new targets in anticancer therapy and can be simulated in some aspects in ground-based facilities.

Role of Hfq in an animal–microbe symbiosis under simulated microgravity conditions

AbstractMicrogravity has a profound impact on the physiology of pathogenic microbes; however, its effects on mutualistic microbes are relatively unknown. To examine the effects of microgravity on those beneficial microbes that associate with animal tissues, we used the symbiosis between the bobtail squidEuprymna scolopesand a motile, luminescent bacterium,Vibrio fischerias a model system. Specifically, we examined the role of Hfq, an RNA-binding protein known to be an important global regulator under space flight conditions, in the squid–vibrio symbiosis under simulated microgravity. To mimic a reduced gravity environment, the symbiotic partners were co-incubated in high-aspect-ratio rotating wall vessel bioreactors and examined at various stages of development. Results indicated that under simulated microgravity,hfqexpression was down-regulated inV. fischeri. A mutant strain defective inhfqshowed no colonization phenotype, indicating that Hfq was not required to initiate the symbiosis with the host squid. However, thehfqmutant showed attenuated levels of apoptotic cell death, a key symbiosis phenotype, within the host light organ suggesting that Hfq does contribute to normal light organ morphogenesis. Results also indicated that simulated microgravity conditions accelerated the onset of cell death in wild-type cells but not in thehfqmutant strains. These data suggest that Hfq plays an important role in the mutualism betweenV. fischeriand its animal host and that its expression can be negatively impacted by simulated microgravity conditions.

Spaceflight Environment Induces Mitochondrial Oxidative Damage in Ocular Tissue

A recent report shows that more than 30% of the astronauts returning from Space Shuttle missions or the International Space Station (ISS) were diagnosed with eye problems that can cause reduced visual acuity. We investigate here whether spaceflight environment-associated retinal damage might be related to oxidative stress-induced mitochondrial apoptosis. Female C57BL/6 mice were flown in the space shuttle Atlantis (STS-135), and within 3-5 h of landing, the spaceflight and ground-control mice, similarly housed in animal enclosure modules (AEMs) were euthanized and their eyes were removed for analysis. Changes in expression of genes involved in oxidative stress, mitochondrial and endothelial cell biology were examined. Apoptosis in the retina was analyzed by caspase-3 immunocytochemical analysis and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Levels of 4-hydroxynonenal (4-HNE) protein, an oxidative specific marker for lipid peroxidation were also measured. Evaluation of spaceflight mice and AEM ground-control mice showed that expression of several genes playing central roles in regulating the mitochondria-associated apoptotic pathway were significantly altered in mouse ocular tissue after spaceflight compared to AEM ground-control mice. In addition, the mRNA levels of several genes, which are responsible for regulating the production of reactive oxygen species were also significantly up-regulated in spaceflight samples compared to AEM ground-control mice. Further more, the level of HNE protein was significantly elevated in the retina after spaceflight compared to controls. Our results also revealed that spaceflight conditions induced significant apoptosis in the retina especially inner nuclear layer (INL) and ganglion cell layer (GCL) compared to AEM ground controls. The data provided the first evidence that spaceflight conditions induce oxidative damage that results in mitochondrial apoptosis in the retina. This data suggest that astronauts may be at increased risk for late retinal degeneration.

Study of the autonomic regulation of blood circulation during a long-term space flight

The five-year experience of experimentation in the autonomic regulation of blood circulation on board the International Space Station is presented. The heart rate variability (HRV) analysis was the basic methodical approach in these investigations. The probabilistic approach to the estimation of the risk of pathology under long-term spaceflight conditions based on HRV analysis is described. The individual type of autonomic regulation was taken into account in the analysis of the results of the investigations. The type of regulation inherent in every cosmonaut under the conditions of weightlessness has been shown to be retained during subsequent flights. New scientific data on the relationship between the character of the adaptive response of the body to spaceflight factors and the individual type of autonomic response have been obtained. Staying in weightlessness has been shown to be connected with the readjustment of regulatory systems and with transition to the zone of prenosological states. Adaptation responses in weightlessness are characterized by the increased tension of the regulatory systems and the preservation of sufficient functional reserves. The mobilization of additional resources is required after returning to earth, due to which the functional reserve of the mechanisms of regulation decreases. Cosmonauts with the vagotonic and normosympathotonic types of autonomic regulation appear to be the most resistant. The knowledge of the type of autonomic regulation allows us to judge the potential response of the cosmonaut to spaceflight factors. The likelihood estimates were calculated, and the risk categories were determined by the results of HRV analysis in the last months of the flight. Three pathology risk groups were identified. In conclusion, the theoretical and applied significance of the experiments was considered.