Results from C-20A science flights (30 October to 2 November 2017) collecting airborne microorganisms in the troposphere and lower stratosphere.

Airborne microorganisms in the upper troposphere and lower stratosphere remain elusive due to a lack of reliable sample collection systems. To address this problem, we designed, installed, and flight-validated a novel Aircraft Bioaerosol Collector (ABC) for NASA's C-20A that can make collections for microbiological research investigations up to altitudes of 13.7 km. Herein we report results from the first set of science flights—four consecutive missions flown over the United States (US) from 30 October to 2 November, 2017. To ascertain how the concentration of airborne bacteria changed across the tropopause, we collected air during aircraft Ascent/Descent (0.3 to 11 km), as well as sustained Cruise altitudes in the lower stratosphere (~12 km). Bioaerosols were captured on DNA-treated gelatinous filters inside a cascade air sampler, then analyzed with molecular and culture-based characterization. Several viable bacterial isolates were recovered from flight altitudes, including Bacillus sp., Micrococcus sp., Arthrobacter sp., and Staphylococcus sp. from Cruise samples and Brachybacterium sp. from Ascent/Descent samples. Using 16S V4 sequencing methods for a culture-independent analysis of bacteria, the average number of total OTUs was 305 for Cruise samples and 276 for Ascent/Descent samples. Some taxa were more abundant in the flight samples than the ground samples, including OTUs from families Lachnospiraceae, Ruminococcaceae and Erysipelotrichaceae as well as the following genera: Clostridium, Mogibacterium, Corynebacterium, Bacteroides, Prevotella, Pseudomonas, and Parabacteroides. Surprisingly, our results revealed a homogeneous distribution of bacteria in the atmosphere up to 12 km. The observation could be due to atmospheric conditions producing similar background aerosols across the western US, as suggested by modeled back trajectories and satellite measurements. However, the influence of aircraft-associated bacterial contaminants could not be fully eliminated and that background signal was reported throughout our dataset. Considering the tremendous engineering challenge of collecting biomass at extreme altitudes where contamination from flight hardware remains an ever-present issue, we note the utility of using the stratosphere as a proving ground for planned life detection missions across the solar system.

Technology demonstration of a nanopore sequencer operating aboard the ISS in comparison with the same sequencer on the ground.

The AirMISR BARC 2001 data were acquired during a flight over the Beltsville Agricultural Research Center (BARC) on July 21, 2001. The Jet Propulsion Laboratory (JPL) in Pasadena, California provided the data. The Airborne Multi-angle Imaging SpectroRadiometer (AirMISR) is an airborne instrument for obtaining multi-angle imagery similar to that of the satellite-borne Multi-angle Imaging SpectroRadiometer (MISR) instrument, which is designed to contribute to studies of the Earth's ecology and climate. AirMISR flies on the NASA ER-2 aircraft. The Jet Propulsion Laboratory in Pasadena, California built the instrument for NASA. Unlike the satellite-borne MISR instrument, which has nine cameras oriented at various angles, AirMISR uses a single camera in a pivoting gimbal mount. A data run by the ER-2 aircraft is divided into nine segments, each with the camera positioned to a MISR look angle. The gimbal rotates between successive segments, such that each segment acquires data over the same area on the ground as the previous segment. This process is repeated until all nine angles of the target area are collected. The swath width, which varies from 11 km in the nadir to 32 km at the most oblique angle, is governed by the camera's instantaneous field-of-view of 7 meters cross-track x 6 meters along-track in the nadir view and 21 meters x 55 meters at the most oblique angle. The along-track image length at each angle is dictated by the timing required to obtain overlap imagery at all angles, and varies from about 9 km in the nadir to 26 km at the most oblique angle. Thus, the nadir image dictates the area of overlap that is obtained from all nine angles. A complete flight run takes approximately 13 minutes. The 9 camera viewing angles are: 0 degrees or nadir 26.1 degrees, fore and aft 45.6 degrees, fore and aft 60.0 degrees, fore and aft 70.5 degrees, fore and aft. For each of the camera angles, images are obtained at 4 spectral bands. The spectral bands can be used to identify vegetation and aerosols, estimate surface reflectance and ocean color studies. The center wavelengths of the 4 spectral bands are: 443 nanometers, blue 555 nanometers, green 670 nanometers, red 865 nanometers, near-infrared Two types of AirMISR data products are available - the Level 1 Radiometric product (L1B1) and the Level 1 Georectified radiance product (L1B2).

The AirMISR BARC 2001 data were acquired during a flight over the Beltsville Agricultural Research Center (BARC) on July 21, 2001. The Jet Propulsion Laboratory (JPL) in Pasadena, California provided the data. The Airborne Multi-angle Imaging SpectroRadiometer (AirMISR) is an airborne instrument for obtaining multi-angle imagery similar to that of the satellite-borne Multi-angle Imaging SpectroRadiometer (MISR) instrument, which is designed to contribute to studies of the Earth's ecology and climate. AirMISR flies on the NASA ER-2 aircraft. The Jet Propulsion Laboratory in Pasadena, California built the instrument for NASA. Unlike the satellite-borne MISR instrument, which has nine cameras oriented at various angles, AirMISR uses a single camera in a pivoting gimbal mount. A data run by the ER-2 aircraft is divided into nine segments, each with the camera positioned to a MISR look angle. The gimbal rotates between successive segments, such that each segment acquires data over the same area on the ground as the previous segment. This process is repeated until all nine angles of the target area are collected. The swath width, which varies from 11 km in the nadir to 32 km at the most oblique angle, is governed by the camera's instantaneous field-of-view of 7 meters cross-track x 6 meters along-track in the nadir view and 21 meters x 55 meters at the most oblique angle. The along-track image length at each angle is dictated by the timing required to obtain overlap imagery at all angles, and varies from about 9 km in the nadir to 26 km at the most oblique angle. Thus, the nadir image dictates the area of overlap that is obtained from all nine angles. A complete flight run takes approximately 13 minutes. The 9 camera viewing angles are: 0 degrees or nadir 26.1 degrees, fore and aft 45.6 degrees, fore and aft 60.0 degrees, fore and aft 70.5 degrees, fore and aft. For each of the camera angles, images are obtained at 4 spectral bands. The spectral bands can be used to identify vegetation and aerosols, estimate surface reflectance and ocean color studies. The center wavelengths of the 4 spectral bands are: 443 nanometers, blue 555 nanometers, green 670 nanometers, red 865 nanometers, near-infrared Two types of AirMISR data products are available - the Level 1 Radiometric product (L1B1) and the Level 1 Georectified radiance product (L1B2).

NASA s Rodent Research (RR) project is playing a critical role in advancing biomedical research on the physiological effects of space environments. Due to the limited resources for conducting biological experiments aboard the International Space Station (ISS) it is imperative to use crew time efficiently while maximizing high-quality science return. NASA s GeneLab project has as its primary objectives to 1) further increase the value of these experiments using a multi-omics systems biology-based approach and 2) disseminate these data without restrictions to the scientific community. The current investigation assessed viability of RNA DNA and protein extracted from archived RR-1 tissue samples for epigenomic transcriptomic and proteomic assays. During the first RR spaceflight experiment a variety of tissue types were harvested from subjects snap-frozen or RNAlater-preserved and then stored at least a year at -80C after return to Earth. They were then prioritized for this investigation based on likelihood of significant scientific value for spaceflight research. All tissues were made available to GeneLab through the bio-specimen sharing program managed by the Ames Life Science Data Archive and included mouse adrenal glands quadriceps gastrocnemius tibialis anterior extensor digitorum longus soleus eye and kidney. We report here protocols for and results of these tissue extractions and thus the feasibility and value of these kinds of omics analyses. In addition to providing additional opportunities for investigation of spaceflight effects on the mouse transcriptome and proteome in new kinds of tissues our results may also be of value to program managers for the prioritization of ISS crew time for rodent research activities.

The AIRMISR_CLAMS_2001 data were acquired during the CLAMS campaign on July 12, July 17, August 1, and August 2 of 2001. The Jet Propulsion Laboratory (JPL) in Pasadena, California provided the data. The Chesapeake Lighthouse and Aircraft Measurements for Satellites (CLAMS) field campaign was held in the summer of 2001 at the CERES Ocean Validation Experiment (COVE) site in the Chesapeake Bay, 20 km east of Virginia Beach. CLAMS is a clear-sky, shortwave closure campaign in conjunction with MISR, CERES, MODIS-Atmospheres and the Global Aerosol Climatology Project (GACP). Its goals were to obtain more accurate broadband fluxes at sea surface and within the atmosphere, space-time variability of spectral BRDF of the sea surface, and aerosol retrievals. The Airborne Multi-angle Imaging SpectroRadiometer (AirMISR) is an airborne instrument for obtaining multi-angle imagery similar to that of the satellite-borne Multi-angle Imaging SpectroRadiometer (MISR) instrument, which is designed to contribute to studies of the Earth's ecology and climate. AirMISR flies on the NASA ER-2 aircraft. The Jet Propulsion Laboratory in Pasadena, California built the instrument for NASA. Unlike the satellite-borne MISR instrument, which has nine cameras oriented at various angles, AirMISR uses a single camera in a pivoting gimbal mount. A data run by the ER-2 aircraft is divided into nine segments, each with the camera positioned to a MISR look angle. The gimbal rotates between successive segments, such that each segment acquires data over the same area on the ground as the previous segment. This process is repeated until all nine angles of the target area are collected. The swath width, which varies from 11 km in the nadir to 32 km at the most oblique angle, is governed by the camera's instantaneous field-of-view of 7 meters cross-track x 6 meters along-track in the nadir view and 21 meters x 55 meters at the most oblique angle. The along-track image length at each angle is dictated by the timing required to obtain overlap imagery at all angles, and varies from about 9 km in the nadir to 26 km at the most oblique angle. Thus, the nadir image dictates the area of overlap that is obtained from all nine angles. A complete flight run takes approximately 13 minutes. The 9 camera viewing angles are: 0 degrees or nadir 26.1 degrees, fore and aft 45.6 degrees, fore and aft 60.0 degrees, fore and aft 70.5 degrees, fore and aft. For each of the camera angles, images are obtained at 4 spectral bands. The spectral bands can be used to identify vegetation and aerosols, estimate surface reflectance and ocean color studies. The center wavelengths of the 4 spectral bands are: 443 nanometers, blue 555 nanometers, green 670 nanometers, red 865 nanometers, near-infrared two types of AirMISR data products are available - the Level 1 Radiometric product (L1B1) and the Level 1 Georectified radiance product (L1B2).

The objective of the Rodent Research-6 (RR-6) study was to evaluate muscle atrophy in mice during spaceflight and to test the efficacy of a novel therapeutic to mitigate muscle wasting. The experiment involved an implantable subcutaneous nanochannel delivery system (nDS; between scapula) which delivered the drug formoterol (FMT; a selective xce xb22 adrenoceptor agonist) over the course of time. To this end a cohort of forty 32-weeks-old female C57BL/6NTac mice were either sham operated or implanted with vehicle or treatment-filled nDS launched in two Transporters (20 mice per Transporter) on SpaceX-13 on December 15 2017. They were transferred to Rodent Habitats onboard the International Space Station (ISS) and maintained in microgravity for 29 days (N=20 Live Animal Return Spaceflight [LAR FLT]) or >50 days (N=20 ISS Terminal Spaceflight [ISS-T FLT]). After 29 days the 20 LAR FLT animals were returned live to back to Earth on January 13 2018. After splashdown the animals were ambulatory on-ground for ~4 days until all subjects were processed during one day of dissections. There were two Basal (BSL) groups of animals sacrificed (LAR BSL & ISS-T BSL; N=20; 40 animals; ~36 weeks old) at Kennedy Space Center (KSC; 12/9/17). LAR BSL animals were dissected and samples were collected upon euthanasia. A Ground Control (GC) group LAR GC mimicked the LAR FLT group which was housed at KSC then shipped alive to Novartis xe2 x80 x99s Facilities where both the LAR FLT and LAR GC groups were processed (~41 weeks old; 1/16/18). All were anesthetized with isoflurane blood samples were obtained by closed-chest cardiac puncture and the animals were euthanized by exsanguination and thoracotomy. The 20 ISS-T FLT mice were anesthetized via intraperitoneal injection of ketamine/xylazine/acepromazine over the course of a four days of dissections (2/6/18 until 2/9/18; 53-56 days after launch; 44 weeks old at time of on-orbit dissections). Blood samples and euthanasia were conducted the same as LAR groups. Following blood draw and hind limb dissection the ISS-T FLT animal carcasses were wrapped in aluminum foil placed in a ziploc bag and placed in storage at -80 xcb x9aC or colder until return. The ISS-T Ground Control (ISS-T GC) (at KSC) followed the same euthanasia timeline methods and preservation. The final processing of frozen ISS-T FLT frozen ISS-T GC and frozen 0-day ISS-T BSL animals were completed at Houston Methodist Research Institute in Houston TX (5/21/18 until 5/24/18). GeneLab received feces from only sham treated animals (no drug treated animals) from the following groups. FLT: LAR (n=9) ISS-T (n=7); GC: LAR (N=7) ISS-T (N=9); BSL: LAR (n=7) ISS-T (n=9). DNA was extracted and analyzed by sequencing using a variety of different targeted and un-targeted metagenome profiling assays.

NASA s Rodent Research (RR) project is playing a critical role in advancing biomedical research on the physiological effects of space environments. Due to the limited resources for conducting biological experiments aboard the International Space Station (ISS) it is imperative to use crew time efficiently while maximizing high-quality science return. NASA s GeneLab project has as its primary objectives to 1) further increase the value of these experiments using a multi-omics systems biology-based approach and 2) disseminate these data without restrictions to the scientific community. The current investigation assessed viability of RNA DNA and protein extracted from archived RR-1 tissue samples for epigenomic transcriptomic and proteomic assays. During the first RR spaceflight experiment a variety of tissue types were harvested from subjects snap-frozen or RNAlater-preserved and then stored at least a year at -80C after return to Earth. They were then prioritized for this investigation based on likelihood of significant scientific value for spaceflight research. All tissues were made available to GeneLab through the bio-specimen sharing program managed by the Ames Life Science Data Archive and included mouse adrenal glands quadriceps gastrocnemius tibialis anterior extensor digitorum longus soleus eye and kidney. We report here protocols for and results of these tissue extractions and thus the feasibility and value of these kinds of omics analyses. In addition to providing additional opportunities for investigation of spaceflight effects on the mouse transcriptome and proteome in new kinds of tissues our results may also be of value to program managers for the prioritization of ISS crew time for rodent research activities.

The BRIC-21 mission was designed to identify the response of Bacillus subtilis to the human spaceflight environment. For this mission samples were grown in rich-medium using the Biological Research in Canister Petri Dish Fixation Units (BRIC-PDFU) spaceflight hardware. B. subtilis spores were inoculated during spaceflight grown at the ambient ISS temperature and frozen in the onboard -80 C freezer prior to returning to Earth. RNA was extracted from samples grown onboard the International Space Station (ISS) and matching Ground Controls for transcriptome analysis.