<abstract>
<title style='display:none'>Abstract</title>
Blue Origin's New Shepard launch vehicle made its first flight above the Kármán Line, returning safely to Earth in November 2015. At the time when this paper is being written (February 2021), the system has conducted 14 flights, including 10 with research and education payloads aboard. More than 100 payloads have exercised a wide range of capabilities and interfaces, from small cubesat-form factor student payloads to large custom payloads of nearly 100 kg. Investigations have spanned a wide range of high-altitude and microgravity research objectives, as well as raising technology readiness level (TRL) on diverse hardware. This paper summarizes New Shepard's payload missions to date, and presents standardized and custom accommodations, both in the shirtsleeve cabin and directly exposed to the space environment.
</abstract>

Research Flights on Blue Origin's New Shepard

<abstract>
<title style='display:none'>Abstract</title>
The increasing availability of flights on suborbital rockets creates new avenues for the study of spaceflight effects on biological systems, particularly of the transitions between hypergravity and microgravity. This paper presents an initial comparison of the responses of Arabidopsis thaliana to suborbital and atmospheric parabolic flights as an important step toward characterizing these emerging suborbital platforms and their effects on biology. Transcriptomic profiling of the response of the Arabidopsis ecotype Wassilewskija (WS) to the aggregate suborbital spaceflight experiences in Blue Origin New Shepard and Virgin Galactic SpaceShipTwo revealed that the transcriptomic load induced by flight differed between the two flights, yet was biologically related to traditional parabolic flight responses. The sku5 skewing mutant and 14-3-3κ:GFP regulatory protein overexpression lines, flown in the Blue Origin and parabolic flights, respectively, each showed altered intra-platform responses compared to WS. An additional parabolic flight using the F-104 Starfighter showed that the response of 14-3-3κ:GFP to flight was modulated in a similar manner to the WS line. Despite the differing genotypes, experimental workflows, flight profiles, and platforms, differential gene expression linked to remodeling of central metabolic processes was commonly observed in the flight responses. However, the timing and directionality of differentially expressed genes involved in the conserved processes differed among the platforms. The processes included carbon and nitrogen metabolism, branched-chain amino acid degradation, and hypoxic responses. The data presented herein highlight the potential for various suborbital platforms to contribute insights into biological responses to spaceflight, and further suggest that in-flight fixation during suborbital experiments will enhance insights into responses during each phase of flight.
</abstract>

Shared Metabolic Remodeling Processes Characterize the Transcriptome of <italic>Arabidopsis thaliana</italic> within Various Suborbital Flight Environments

<abstract><title style='display:none'>Abstract</title>In this work, we report on the construction, training and functional assessment of an electronic nose (called ‘E-Nose’) that is capable of monitoring the microbial contamination onboard space ships under microgravity conditions. To this end, a commercial electronic nose was modified to allow for the sampling of microbial volatile organic compounds (MVOCs) emitted from relevant bacterial and fungi species. Training of the modified ‘E-Nose’ was performed by establishing an MVOC database consisting of two Gram-positive bacteria strains (Bacillus subtilis and Staphylococcus warneri) and two fungi strains (Aspergillus versicolor and Penicillium expansum). All these strains are known to exist onboard the International Space Station (ISS) and to form important parts of its microbial contamination. All cultures were grown on four kinds of structural materials also in use onboard the ISS. The MVOCs emitted during the different growth phases of these cultures were monitored with an array of ten different metal oxide gas sensors inside the ‘E-Nose’. Principal component analysis of the array data revealed that B. subtilis and S. warneri form separate clusters in an optimized score plot, while the two fungi strains of A. versicolor and P. expansum form a large common cluster, well discriminated against to the bacteria clusters.</abstract>

Detection of Microorganisms with an Electronic Nose for Application under Microgravity Conditions

<abstract><title style='display:none'>Abstract</title>High-altitude balloons (HABs) present a valuable and cost-effective tool for educators and students to access the conditions that are analogous to space and extraterrestrial environments in the Earth’s upper atmosphere. Historically, HABs have been used for meteorological measurements, observation, sampling of aerosols, and exposure of samples to upper atmosphere environments. The Earth’s stratosphere allows researchers access to a unique combination of wideband solar radiation, extreme cold, rarefied air, low humidity, and acute ionizing radiation—conditions that are relevant to space biology research. Here, we describe a reproducible payload for a HAB mission that can be constructed, launched, and retrieved for about $3,000. This general standard operating procedure can be used by educators, community scientists, and research teams working with limited resources.</abstract>

A High-Altitude Balloon Platform for Space Life Sciences Education

<abstract><title style='display:none'>Abstract</title>This work investigates the suitability of membrane aerated biological reactors (MABRs) for biological treatment of a space-based waste stream consisting of urine, hygiene/grey water, and humidity condensate within an overall water recycling system. Water represents a critical limiting factor for human habitation and travel within space; thus, water recycling systems are essential. Biological treatment of wastewater provides a more efficient sustainable means of stabilizing the waste stream within water recycling system architectures in comparison to current chemical stabilization processes that utilize harsh chemicals, which represent both a hazardous and an unsustainable approach. To assess the capabilities of MABRs for providing microgravity compatible biological treatment and verify long duration operation and integration with desalination processes, two full-scale MABR systems were challenged with various loading rates and operational scenarios during sustained operation for over 1 year. The MABRs were able to maintain 196 g-C/m3-d and 194 g-N/m3-d volumetric conversion rates. Additionally the systems were able to handle intermittent loading and recover rapidly from system hibernation periods of up to 27 days. Overall, the use of MABRs within a wastewater treatment system architecture provides several potential benefits including minimizing the use of toxic chemical pretreatment solutions and providing an effluent solution that is easier to desalinate and dewater.</abstract>

Assessment of Membrane-Aerated Biological Reactors (MABRs) for Integration into Space-Based Water Recycling System Architectures

<abstract><title style='display:none'>Abstract</title>Increasing efforts to move into space have driven the need for new facilities that are capable of simulating weightlessness and other space gravity conditions on Earth. Simulation of weightlessness/<italic>microgravity</italic> (approximately 10−6<italic>g</italic>) is conducted in different earthbound and flight-based facilities, often with poor availability. Other conditions such as lunar or Martian gravity with their partial Earth gravity/<italic>hypogravity</italic> cannot be performed at a large scale for scientific research on Earth. For multiple Earth gravity/<italic>hypergravity</italic>, simulation centrifuges are available, but they do not allow the possibility of abrupt acceleration changes. To support this wide range of conditions, a new technique is being developed to combine all of these requirements into a single drop tower facility. Currently under construction, the <italic>Einstein-Elevator</italic> of the Hannover Institute of Technology at the Leibniz Universität Hannover is an earthbound tool created for simulating micro-, hypo-, and hypergravity research with a high repetition rate. The facility will be capable of performing 100 experiments per day (8-h work shift), each creating 4 s of microgravity. For the first time, statistics can be applied in experiments under space gravity conditions at favorable costs and short mission times. The Einstein-Elevator offers room for large experiments with a diameter up to 1.7 m and a height up to 2 m as well as weights up to 1,000 kg. To perform larger experiments under different gravitational conditions, it was necessary to develop an innovative drive and guide concept. The Einstein-Elevator will be available for general research under different gravity conditions from 2018 onward.</abstract>

Einstein-Elevator: A New Facility for Research from μ<italic>g</italic> to 5 <italic>g</italic>

<abstract>
<title style='display:none'>Abstract</title>
As human settlements expand to lunar and Martian bases, optimizing food production in these environments becomes crucial. This study investigates the use of macroalgae, specifically <italic>Ulva lactuca</italic> L., as an affordable, sustainable approach for seed priming to enhance germination in extraterrestrial soils. The focus was on the germination and growth of <italic>Capsicum annuum</italic> L. (pepper), <italic>Lactuca sativa</italic> L. (lettuce), <italic>Cicer arietinum</italic> L. (chickpea), and <italic>Pisum sativum</italic> L. (pea) in simulated Martian and lunar regolith. Two concentrations of <italic>U. lactuca</italic> powder (0.2 and 0.4 g · L−1) were tested under controlled conditions. The study also conducted a qualitative chemical analysis of <italic>U. lactuca</italic> to identify bioactive components essential for phytohormone formation. The germination and emergence rates of the seeds in the lunar regolith were higher than those in the Martian regolith. Martian regolith's optimal treatment for pea and chickpea seed germination was 0.2 g · L−1, which also favored seedling emergence. In the lunar regolith, optimal germination rates for pea seeds were observed with both treatments and chickpea seeds. The germination percentage of lettuce seeds in the lunar regolith was higher than the control, with 0.2 g · L−1, while there was no significant difference for the other seeds. The study recommends the application of <italic>U. lactuca</italic> powder as an effective biostimulant for the examined cultivars due to the presence of plant growth regulators (PGRs) that enhance germination and seedling emergence under challenging conditions.
</abstract>

Instituto de Estudos do Mar Almirante Paulo Moreira (IEAPM) and Universidade Federal Fluminense (UFF)

Seed Priming with <italic>Ulva lactuca L</italic>. in Cultivars Grown in Martian and Lunar Regolith Analogues

<abstract>
<title style='display:none'>Abstract</title>
<italic>Enterobacter cloacae</italic>, a gram-negative bacterium commonly found in the human gut microbiota, poses potential health risks to astronauts in the unique environment of space flight. This study investigated the effects of payload shielding on <italic>E. cloacae</italic> in a short-duration, student-initiated, weather balloon flight experiment. Faraday fabric-based payload shielding did not impact the viability of the balloon flight samples. However, murine macrophage infection assays showed that shielded balloon flight <italic>E. cloacae</italic> had significantly improved intracellular survival compared to unshielded <italic>E. cloacae</italic>. Proteomic analysis demonstrated distinct profiles in shielded and unshielded samples, with a differential abundance of proteins involved in diverse biological processes. Specifically, decreased abundance of proteins involved in chemotaxis, DNA repair, replication, transcription, peptidoglycan synthesis, and proteolysis were observed in the Faraday fabric-based payload-shielded samples. In contrast, proteins associated with protein translation, transport, tricarboxylic acid cycle, fatty acid biosynthesis, and amino acid metabolism were increased in shielded conditions. This experiment provides a framework for which future long-duration balloon flight experiments can be designed, and the findings provide initial insights into the impact of payload shielding on <italic>E. cloacae</italic> physiology. Understanding the impact of the stratosphere on human gut microbiota is important for preserving human health during future space flight missions.
</abstract>

Impact of payload shielding on <italic>Enterobacter cloacae</italic> viability and proteomic profile: Insights from a stratospheric weather balloon flight experiment

<abstract>
<title style='display:none'>Abstract</title>
As considerations are being made for the limitations and safety of long-term human spaceflight, the vasculature is important given its connection to and impact on numerous organ systems. As a major constituent of blood vessels, vascular smooth muscle cells are of interest due to their influence over vascular tone and function. Additionally, vascular smooth muscle cells are responsive to pressure and flow changes. Therefore, alterations in these parameters under conditions of microgravity can be functionally disruptive. As such, here we review and discuss the existing literature that assesses the effects of microgravity, both actual and simulated, on smooth muscle cells. This includes the various methods for achieving or simulating microgravity, the animal models or cells used, and the various durations of microgravity assessed. We also discuss the various reported findings in the field, which include changes to cell proliferation, gene expression and phenotypic shifts, and renin-angiotensin-aldosterone system (RAAS), nitric oxide synthase (NOS), and Ca2+ signaling. Additionally, we briefly summarize the literature on smooth muscle tissue engineering in microgravity as well as considerations of radiation as another key component of spaceflight to contextualize spaceflight experiments, which by their nature include radiation exposure. Finally, we provide general recommendations based on the existing literature's focus and limitations.
</abstract>

The Effects of Simulated and Real Microgravity on Vascular Smooth Muscle Cells

<abstract>
<title style='display:none'>Abstract</title>
This short article describes the major findings from the CVB experiment performed in the LMM on the International Space Station from 2010–2012. CVB was the first experiment to run in the new facility and focused on understanding the heat transfer and fluid mechanics occurring inside a wickless miniature heat pipe. The LMM was used to map the location of the vapor-liquid interface inside the device and to measure the film thickness profile on the walls of the device. Several interesting and unexpected phenomena were observed in microgravity including flooding of the heater end with liquid as the heat input increased, explosive nucleation of vapor bubbles at the heater end in the shortest version of the heat pipe tested, condensation on highly superheated surfaces, and the spontaneous formation of rip currents as the device tried to enhance the contact line area available for evaporation of the liquid.
</abstract>

Constrained Vapor Bubble Experiment (CVB) in the Light Microscopy Module (LMM)

<abstract>
<title style='display:none'>Abstract</title>
Accelerated aging in space is detrimental to long-term space missions. The environmental conditions in space (e.g., microgravity and radiation) cause harmful effects similar to those seen during aging. As the mechanistic pathways underlying accelerated aging in spaceflight are not fully understood, the identification of critical targets for promoting longevity in spaceflight remains challenging. We analyzed genomics data from the GLDS-207 project to identify potential targets related to longevity. Analysis of RNA-seq data from four Drosophila variants using the GeneLab Galaxy platform indicated that spaceflight significantly affected differential gene expression in the heads of flies, specifically in the seizure (sei) mutant, which alters the voltage gated potassium channels in the cell membrane. Spaceflight induced a significant decrease in the expression of the retinal degeneration A gene (rdgA) in mutant flies that survived the 30-day space mission. This gene encodes for the protein diacylglycerol kinase (DGK), which modulates the activation of the mechanistic target of the rapamycin (mTOR) signaling pathway, known to negatively regulate aging. Therefore, DGK may be a potential target for promoting longevity in space conditions. Further investigation of the effects of decreased rdgA expression on the lifespan of other organisms under spaceflight conditions will clarify the role of DGK in promoting longevity.
</abstract>

Diacylglycerol kinase is downregulated in the Drosophila Seizure Mutant during Spaceflight

<abstract>
<title style='display:none'>Abstract</title>
NASA's recent emphasis on human exploration of the Moon and, ultimately, Mars necessitates a transition from a focus of its research in the biological sciences from Low Earth Orbit (LEO) to platforms beyond LEO. Fundamental research questions need to be addressed to enable humans to thrive in deep space. Work beyond LEO necessitates a shift in technology and the utilization of organisms in autonomous experiments, especially in the near term. The Beyond LEO Instrumentation &amp; Science Series Science Working Group (BLISS-SWG) was established to provide NASA's Space Biology Program input on its strategy for developing research priorities and tools for exploration beyond LEO. Here, we present an abridged version of the first annual report of the BLISS-SWG, which is publicly available on the NASA Technical Reports Server. Seven priority areas and pertinent research questions were identified for research beyond LEO in the coming 2–5 years. Appropriate experimental organisms and technology development needs for research addressing these questions are summarized. The BLISS-SWG aims for this review to serve as a resource for the space biology and science and engineering communities as they develop research to understand risks and mitigation strategies for deep-space stressors on human crew, plants, and their microbiomes.
</abstract>

Strategies, Research Priorities, and Challenges for the Exploration of Space Beyond Low Earth Orbit

<abstract><title style='display:none'>Abstract</title>The first Veggie plant growth chamber was installed on the International Space Station in 2014. Crop plants can be grown in Veggie using plant pillows, small rooting packets that contain substrate, fertilizer, and germination wicks along with attached seeds. The pillows were designed to interface with the Veggie root mat reservoir watering system to provide a capillary water column to growing plants. In preparation for flight, methods of arcillite substrate washing, autoclaving, and drying were established to reduce dust and to provide a dry sterile substrate. A controlled released fertilizer mixed into arcillite substrate provides nutrition for plant growth. Methods of seed surface sterilization were tested for both germination and microbial contamination, and the optimum methods were determined for candidate flight crops. Plant pillows were prepared for flight by cutting and inserting germination wicks, filling with the substrate/fertilizer mix, and sewing closed. Following pillow filling, seeds were attached to the wicks, and the pillows were packaged for flight. Pillow preparation methods have been successfully tested in the VEG-01 hardware validation tests on the International Space Station with ‘Outredgeous’ lettuce and ‘Profusion’ zinnia, and in the VEG-03 test, using ‘Outredgeous’ lettuce and ‘Tokyo bekana’ Chinese cabbage.</abstract>

Plant Pillow Preparation for the Veggie Plant Growth System on the International Space Station

<abstract>
<title style='display:none'>ABSTRACT</title>
Exposure to total-body radiation induces hematological changes, which can detriment one’s immune response to wounds and infection. Here, the decreases in blood cell counts after acute radiation doses of γ-ray or proton radiation exposure, at the doses and dose-rates expected during a solar particle event (SPE), are reported in the ferret model system. Following the exposure to γ-ray or proton radiation, the ferret peripheral total white blood cell (WBC) and lymphocyte counts decreased whereas neutrophil count increased within 3 hours. At 48 hours after irradiation, the WBC, neutrophil, and lymphocyte counts decreased in a dose-dependent manner but were not significantly affected by the radiation type (γ-rays verses protons) or dose rate (0.5 Gy/minute verses 0.5 Gy/hour). The loss of these blood cells could accompany and contribute to the physiological symptoms of the acute radiation syndrome (ARS).
</abstract>

The Effects of Gamma and Proton Radiation Exposure on Hematopoietic Cell Counts in the Ferret Model

<abstract>
<title style='display:none'>ABSTRACT</title>
Plants will be an important component of off-Earth life support systems for food production and atmosphere recycling. “Veggie” is a small vegetable production unit designed for space flight, with a passive water delivery system. Plants can be grown in Veggie using small bags with a wicking surface containing media and fertilizer, i.e., pillows. Pillows planted with seeds can be placed on the wicking surface of the Veggie reservoir and water will wick throughout the media. Multiple small salad and herb species were grown in Veggie analog conditions using both commercial peat-based media and arcillite. Biometric measurements and microbial loads were assessed. Some species grew better in a particular media, but no general trends were apparent. Lettuce plants grew best in the blends of the peat-based and arcillite media. Microbial counts were lower on plants grown in arcillite. Four media types (peat-based mix, arcillite, and blends of the two) were tested in the rooting pillows; tests included Chinese cabbage, Swiss chard, lettuce, snow pea, and radish. Most species grew best in blends of the commercial mix and arcillite. Edible biomass production varied from 3.5-8 grams dry mass/m2/day with lettuce having the lowest biomass and Chinese cabbage highest. Radish plants showed an increasing percentage of partitioning to edible roots with increasing arcillite in the media. Pillows appear to offer a simple, effective strategy for containing rooting media and avoiding free water while growing plants in the Veggie hardware.
</abstract>

Preliminary Species and Media Selection for the Veggie Space Hardware

<abstract><title style='display:none'>Abstract</title>Despite centuries of scientific balloon flights, only a handful of experiments have produced biologically relevant results. Yet unlike orbital spaceflight, it is much faster and cheaper to conduct biology research with balloons, sending specimens to the near space environment of Earth's stratosphere. Samples can be loaded the morning of a launch and sometimes returned to the laboratory within one day after flying. The National Aeronautics and Space Administration (NASA) flies large unmanned scientific balloons from all over the globe, with missions ranging from hours to weeks in duration. A payload in the middle portion of the stratosphere (~35 km above sea level) will be exposed to an environment similar to the surface of Mars—temperatures generally around −36°C, atmospheric pressure at a thin 1 kPa, relative humidity levels &lt;1%, and harsh illumination of ultraviolet (UV) and cosmic radiation levels (about 100 W/m2 and 0.1 mGy/d, respectively)—that can be obtained nowhere else on the surface of the Earth, including environmental chambers and particle accelerator facilities attempting to simulate space radiation effects. Considering the operational advantages of ballooning and the fidelity of space-like stressors in the stratosphere, researchers in aerobiology, astrobiology, and space biology can benefit from balloon flight experiments as an intermediary step on the extraterrestrial continuum (i.e., ground, low Earth orbit, and deep space studies). Our review targets biologists with no background or experience in scientific ballooning. We will provide an overview of large balloon operations, biology topics that can be uniquely addressed in the stratosphere, and a roadmap for developing payloads to fly with NASA.</abstract>

Ballooning for Biologists: Mission Essentials for Flying Life Science Experiments to Near Space on NASA Large Scientific Balloons

<abstract><title style='display:none'>ABSTRACT</title>Space travel beyond the Earth’s protective magnetosphere risks exposing astronauts to ionizing radiation, such as that generated during a solar particle event (SPE). Ionizing radiation has well documented effects on blood cells and it is generally assumed that these effects contribute to the hematopoietic syndrome (HS), observed in animals and humans, following exposure to total body irradiation (TBI). The purpose of the current study was to assess the role of gender on the effects of gamma radiation on blood cells. C3H/HeN mice were irradiated with a 137Cs gamma source. Radiation had similar effects on white blood cells (WBCs), lymphocytes, and granulocytes in male and female C3H/HeN mice, while red blood cell (RBC) counts and hematocrit values remained stable following radiation exposure. Non-irradiated male mice had 13% higher platelet counts, compared with their female counterparts, and showed enhanced recovery of platelets on day 16 following radiation exposure. Hence, gender differences influence the response of platelets to TBI exposure.</abstract>

Effect of Gender on the Radiation Sensitivity of Murine Blood Cells

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Gravitational and Space Research

 Gravitational and Space Research is the main journal of the American Society for Gravitational and Space Research (ASGSR) and publishes quality, peer-reviewed manuscripts...

 Gravitational and Space Research is the main journal of the American Society for Gravitational and Space Research (ASGSR) and...

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