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Investigation of Murine T-Cells and Cancer Cells under Thermal Stressors and 2D Slow Rotating System Effects as a Testbed for Suborbital Flights

Abstract

Research indicates that exposure to microgravity leads to immune system dysregulation. However, there is a lack of clear evidence on the specific reasons and precise mechanisms accounting for these immune system changes. Past studies investigating space travel-induced alterations in immunological parameters report many conflicting results, explained by the role of certain confounders, such as cosmic radiation, individual body environment, or differences in experimental design. To minimize the variability in results and to eliminate some technical challenges, we advocate conducting thorough feasibility studies prior to actual suborbital or orbital space experiments. We show how exposure to suborbital flight stressors and the use of a two-dimensional slow rotating device affect T-cells and cancer cells survivability. To enhance T-cell activation and viability, we primed them alone or in combination with IL-2 and IL-12 cytokines. Viability of T-cells was assessed before, during the experiment, and at the end of the experiment for which T-cells were counted every day for the last 4 days to allow the cells to form clear structures and do not disturb their evolution into various geometries. The slow rotating device could be considered a good system to perform T-cell activation studies and develop cell aggregates for various types of cells that react differently to thermal stressors.

Open access
Effect of macromolecular mass transport in microgravity protein crystallization

Abstract

To investigate the effect of macromolecular transport and the incorporation of protein aggregate impurities in growing crystals, experiments were performed on the International Space Station (ISS) and compared with control experiments performed in a 1G laboratory environment. Crystal growth experiments for hen egg-white lysozyme (HEWL) and Plasmodium falciparum glutathione S-transferase (PfGST) were monitored using the ISS Light Microscopy Module (LMM). Experiments were performed applying the liquid–liquid counter diffusion crystallization method using rectangular, optically transparent capillaries. To analyze the quantity of impurity incorporated into growing crystals, stable fluorescently labeled protein aggregates were prepared and subsequently added at different percent concentrations to nonlabeled monomeric protein suspensions. For HEWL, a covalent cross-linked HEWL dimer was fluorescently labeled, and for PfGST, a stable tetramer was prepared. Crystallization solutions containing different protein aggregate ratios were prepared. The frozen samples were launched on 19.02.2017 via SpaceX-10 mission and immediately transferred to a -80°C freezer on the ISS. Two series of crystallization experiments were performed on ISS, one during 26.02.2017 to 10.03.2017 and a second during 16.06.2017 to 23.06.2017. A comparison of crystal growth rate and size showed different calculated average growth rates as well as different dimensions for crystals growing in different positions along the capillary. The effect of macromolecular mass transport on crystal growth in microgravity was experimentally calculated. In parallel, the percentage of incorporated fluorescent aggregate into the crystals was monitored utilizing the fluorescent LMM and ground-based fluorescent microscopes.

Open access
Impact of g-Load Shift on Temporal Expression Pattern of Apoptosis-linked Proteins in the Rat Mammary Gland

Abstract

Alteration in gravitational load impacts homeorhetic response in rat dams which affects neonatal pup survival. However, the effects of hypergravity (HG) exposure on the abundance of apoptosis-associated proteins in mammary epithelial cells (MECs) have not been characterized. Therefore, we examined whether chronic exposure to HG from midpregnancy alters the abundance of proapoptotic proteins in MECs during the late pregnancy and early lactation. A group of pregnant Sprague Dawley rats were exposed to either HG (2g) or normo-gravity (1g: stationary control [SC]) from days 11 to 20 of gestation (G20). Another set of animals were investigated from day 11 of pregnancy through days 1 and 3 (P1 and P3, respectively) postpartum. Quantitative (pixels [px]/lobule) immunohistochemistry at G20 of Cleaved Caspase-3 (CC-3), Tumor Protein p53 (P53), and vitamin D receptor (VDR) revealed that all the three proteins were increased (p<0.01) in HG rats compared to SC animals. At P1, the HG group had twofold higher (p<0.001) expression of CC-3 relative to the SC group. Approximately, 50% (p<0.001) more VDR was detected in the HG cohorts than SC at P3. These results suggest that a shift in g-load upregulates the expression of key proapoptotic proteins during the pregnancy-to-lactation transition in the rat MECs.

Open access
Musculoskeletal Outcomes from Chronic High-Speed High-Impact Resistive Exercise

Abstract

Subjects (n=13) did 30 workouts with their left leg on an Inertial Exercise Trainer (IET), while their right leg served as an untreated control. Before and after the 30 workouts, they underwent isokinetic strength tests (knee and ankle extensors of both legs) whose peak torque (PT), time to PT (TTPT), and rate of torque development (RTD) values were each analyzed with 2(leg)×2(time)×3(velocity) analysis of variances (ANOVAs), with repeated measures per independent variable. Peak force (PF) and total work (TW) data were measured from each IET workout, and they represent time course strength changes produced by our exercise intervention. PF and TW values for the three IET exercises that comprised each workout were each analyzed with one-way ANOVAs with time as the independent variable. Results included significant ankle and knee extensor PT increases, whereby the left leg achieved higher values at posttesting, but there were no significant TTPT changes and a time effect for ankle extensor RTD. Our data show that PF and TW each had significant increases over time, with the latter exhibiting greater gains over the 30-workout intervention. Our results imply that the IET yields strength gains over time comparable to standard resistive exercise hardware.

Open access
Challenges of ERAU’s First Suborbital Flight Aboard Blue Origin’s New Shepard M7 for the Cell Research Experiment In Microgravity (CRExIM)

Abstract

Cell Research Experiment In Microgravity (CRExIM) was launched aboard Blue Origin’s New Shepard suborbital vehicle on Tuesday, December 12, 2017, from the West Texas Launch Site in Van Horn, Texas. One of the aims of this science experiment was to assess the effects of microgravity on murine T-cells during suborbital flight. These cells were placed in a NanoLab with a data logger that sensed the acceleration, temperature, and relative humidity during preflight, flight, and postflight operations. Some discrepancies in sensor measurement were noticed, and these errors were attributed partly to the difference in sampling rates and partly to the different locations of the sensors, which made it difficult to obtain highly accurate measurements of the accelerations and to correlate both sets of data. This paper discusses the setbacks and lessons learned, which made our team find new alternatives while meeting all milestones as mandated by NanoRacks and Blue Origin. This manuscript highlights these alternatives that led to the success of the mission and gives recommendations that will enable customers to alleviate some of these challenges in future flights.

Open access