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Novel Approaches to Cellular Transplantation from the US Space Program

Research in the treatment of type I diabetes is entering a new era that takes advantage of our knowledge in an ever increasing variety of scientific disciplines. Some may originate from very diverse sources, one of which is the Space Program at National Aeronautics and Space Administration (NASA). The Space Program contributes to diabetes-related research in several treatment modalities. As an ongoing effort for medical monitoring of personnel involved in space exploration activities NASA and the extramural scientific community investigate strategies for noninvasive estimation of blood glucose levels. Part of the effort in the space protein crystal growth program is high-resolution structural analysis insulin as a means to better understand the interaction with its receptor and with host immune components and as a basis for rational design of a "better" insulin molecule. The Space Program is also developing laser technology for potential early cataract detection as well as a noninvasive analyses for addressing preclinical diabetic retinopathy. Finally, NASA developed an exciting cell culture system that affords some unique advantages in the propagation and maintenance of mammalian cells in vitro. The cell culture system was originally designed to maintain cell suspensions with a minimum of hydrodynamic and mechanical sheer while awaiting launch into microgravity. Currently the commercially available NASA bioreactor (Synthecon, Inc., Houston, TX) is used as a research tool in basic and applied cell biology. In recent years there is continued strong interest in cellular transplantation as treatment for type I diabetes. The advantages are the potential for successful long-term amelioration and a minimum risk for morbidity in the event of rejection of the transplanted cells. The pathway to successful application of this strategy is accompanied by several substantial hurdles: (1) isolation and propagation of a suitable uniform donor cell population; (2) management of host immune rejection; (3) protection from the autoimmune component of the disease; and (4) anatomic placement of the engrafted cells that permits timely response to blood sugar levels as well as effective release and deployment of insulin. Bioreactor technology may provide some critical advances for surmounting some of these scientific hurdles. The NASA bioreactor is a horizontally rotating cylinder that is completely filled with culture medium. Gaseous exchange is maintained by a concentric cylinder of permeable silicon. In slow rotation (15-25 rpm) particles of small mass such as cells and tissue aggregates remain suspended in the rotating body of fluid. This novel approach suspends cells without stirring thus, allowing objects of different size mass to colocate and interact in a very low shear environment. In fact, analysis of the forces acting on individual cells in the rotating bioreactor reveals that the cells are continuously falling through the fluid medium. The conditions in the bioreactor permit assembly of cells into aggregates. three-dimensional tissue growth, synthesis of intercellular matrix,10 differentiation, and some sinusoid formations that may serve as a surrogate vasculature for larger tissue segments.

Pellis, Neal R.↗

NASA-DoD Lead-Free Electronics Project. DRAFT Joint Test Report

The use of conventional tin-lead (SnPb) in circuit board manufacturing is under ever-increasing political scrutiny due to increasing regulations concerning lead. The "Restriction of Hazardous Substances" (RoHS) directive enacted by the European Union (EU) and a pact between the United States National Electronics Manufacturing Initiative (NEMI), Europe's Soldertec at Tin Technology Ltd. and the Japan Electronics and Information Technology Industries Association (JEITA) are just two examples where worldwide legislative actions and partnerships/agreements are affecting the electronics industry. As a result, many global commercial-grade electronic component suppliers are initiating efforts to transition to lead-free (Pb-free) in order to retain their worldwide market. Pb-free components are likely to find their way into the inventory of aerospace or military assembly processes under current government acquisition reform initiatives. Inventories "contaminated" by Pb-free will result in increased risks associated with the manufacturing, product reliability, and subsequent repair of aerospace and military electronic systems. Although electronics for military and aerospace applications are not included in the RoHS legislation, engineers are beginning to find that the commercial industry's move towards RoHS compliance has affected their supply chain and changed their parts. Most parts suppliers plan to phase out their non-compliant, leaded production and many have already done so. As a result, the ability to find leaded components is getting harder and harder. Some buyers are now attempting to acquire the remaining SnPb inventory, if it's not already obsolete. Original Equipment Manufacturers (OEMs), depots, and support contractors have to be prepared to deal with an electronics supply chain that increasingly provides more and more parts with Pb-free finishes-some labeled no differently than their Pb counterparts-while at the same time providing the traditional Pb parts. The longer the transition period, the greater the likelihood of Pb-free parts inadvertently being mixed with Pb parts and ending up on what are supposed to be Pb systems. As a result, OEMs, depots, and support contractors need to take action now to either abate the influx of Pb-free parts, or accept it and deal with the likely interim consequences of reduced reliability due to a wide variety of matters, such as Pb contamination, high temperature incompatibility, and tin whiskering. Allowance of Pb-free components produces one of the greatest risks to the reliability of a weapon system. This is due to new and poorly understood failure mechanisms, as well as unknown long-term reliability. If the decision is made to consciously allow Pb-free solder and component finishes into SnPb electronics, additional effort (and cost) will be required to make the significant number of changes to drawings and task order procedures. This project is a follow-on effort to the Joint Council on Aging Aircraft/Joint Group on Pollution Prevention (JCAA/JG-PP) Pb-free Solder Project which was the first group to test the reliability of Pb-free solder joints against the requirements of the aerospace and military community.

Kessel, Kurt↗

The Assembly, Test, and Integration of LOFTID (Low-Earth Orbit Flight Test of an Inflatable Decelera-tor)

R.J. Bodkin Biography Mr. Bodkin worked in industry for a rapid prototype company focusing on UAVs and manned experimental aircraft. Later he served as the Inflation System Lead on IRVE-II and 3 and the Re-Entry Vehicle Lead for LOFTID at NASA Langley Research Center. Introduction: The Low-Earth Orbit Flight Test of an Inflatable Decelerator (LOFTID), developed in partnership with United Launch Alliance (ULA) and flown in conjunction with the National Oceanic and Atmospheric Administration (NOAA) Joint Polar Satellite System-2 (JPSS-2) satellite, demonstrated Hypersonic Inflatable Aerodynamic Decelerator (HIAD) technology has progressed and is ready for mission infusion. LOFTID’s success demonstrates that aeroshells are not limited to the internal diame-ter of the launch vehicle payload fairing, allowing larger payloads to be deployed to the surfaces of planetary bodies with atmospheres. The challenges of assembling, integrating, and testing this revolutionary spacecraft will be dis-cussed as well as issues associated with doing this with a fixed launch date the project did not control. Assembly: Because LOFTID flew as a rideshare partner with JPSS-2, it was constrained with addi-tional schedule, milestone, and technical require-ments that were beyond the project’s control. As-sembly of the LOFTID hardware was challenged with the normal mechanical fit issues while also having to navigate the SARS-COVID-II pandemic. Challenges ranged from availability of team per-sonnel required on-site for vehicle assembly to dif-ficulties associated with team collaboration while working remotely and increased costs and lead times of components due to supply chain con-straints. Numerous additional challenges cascaded from the additional time required. Integration: LOFTID flew as a secondary pay-load to JPSS-2 in a mission-unique configuration, directly under JPSS-2 primary payload, inside the Payload Adapter that integrated JPSS-2 to the Atlas V launch vehicle. A mission unique Payload Adapt-er Separation System (PASS) was required to sepa-rate the Payload Adapter from the Launch Vehicle prior to the start of the LOFTID flight demonstra-tion. Development of this system was challenging due to a shortened development schedule resulting from the iterative nature of Payload Adapter devel-opment with the partners at ULA. Preparations to integrate the main segments of the LOFTID vehicle posed unique challenges of having to accommodate issues with a fixed launch date that led to some cre-ative solutions to the integration. The partnership agreement with ULA and JPSS-2 resulted in a mass simulator designed to be installed late in the inte-gration in the event the LOFTID vehicle was not ready in time.. Test: LOFTID testing was carried out in several phases. Some components were tested at the com-ponent level, others at the sub-system levels and then finally the integrated vehicle level. This culmi-nated with the Complete Systems Test (CST) per-formed in a vacuum chamber as one of the final checkouts prior to disassembly for re-packing of the aeroshell. CST challenges will be discussed as well as obstacles encountered post-CST. After CST, the vehicle was disassembled so the HIAD could be repacked, and the vehicle was reassembled for ac-ceptance vibration testing. Testing concluded with the fully assembled vehicle being shipped to the launch site for final testing and integrations with the Payload Adapter to JPSS-2 for launch and opera-tions. Conclusion: The challenges posed by the AI&T for LOFTID could inform the planetary community of some of the opportunities and challenges of de-veloping technologies on a rideshare with a rela-tively small budget.

R.J. Bodkin↗

ISS External Microorganisms: A Planetary Protection Experiment to Inform Requirements for Crewed Missions to Mars

We have developed, tested, and flown a caddy capable of collecting aseptic samples from external surfaces of the ISS (International Space Station). The sampling caddy is certified for use during US EVA (extra vehicular activity) and was launched to ISS in the summer of 2023. We are scheduled to collect samples from 6 locations outside ISS during an EVA in May of 2024. We will freeze these samples at -80°C on orbit and return them to Earth. We will then extract and sequence any DNA collected during the EVA using next generation sequencing technologies to characterize the community composition and function of each sample. Measuring the type and quantity of microbes present on the exterior of ISS will allow us to address knowledge gap 2B, “Acceptable levels of microbial/organic releases from humans and support systems” described in a 2019 COSPAR report. Collecting data about microbial release from current crewed vehicles will inform requirements for acceptable leak rates for future crewed missions to Mars. The sampling kit consists of eight commercially available, sterile, DNA free, macrofoam swabs ( 23 mm. diameter ) installed in custom aluminum end effectors. Each end effector is housed in and individual aluminum canister. Each canister contains a 0.2 μm Teflon filter to allow the interior volume to accommodate pressure changes without permitting microbial contaminants to enter the sterile interior volume. A handle repurposed from the space shuttle tile repair kit is used to remove the end effector from the sample canister, collect a sample by swabbing a surface and then replace the end effector in its canister. The canisters and end effectors were cleaned and assembled on Earth. Prior to installing the sterile swab the canisters and end effectors were sterilized in an autoclave at 134°C, 215 kPA, for 7 min.. The final assembly occurred in a sterilized class II biosafety cabinet. We will collect six samples from the 1) airlock vestibule, 2) airlock thermal cover, 3) a gap in the micrometeorite shielding near the airlock, 4) a handrail near the airlock, 5) the CDRA (Carbon Dioxide Removal Assembly) vent, and 6) the VES (Vacuum Exhaust System) vent. The remaining two swabs will be reserved as controls. One swab will be exposed during the EVA without touching any surfaces to act as a blank. The final swab will remain sealed until the entire sampling kit is returned to earth. Based on previously published results from the Russian segment, we hypothesize that there will be detectable microbes at some or all of these locations. Ground-based testing of this sampling caddy confirms that the swabs remain sterile as the canisters transition in and out of vacuum. We were able to retrieve, viable bacterial and fungal cells as well as DNA from samples collected from US space suits during vacuum chamber tests lasting as long as seven hours. Based on these results and feedback from the test subjects the sampling caddy was modified to improve ergonomics and meet US EVA safety requirements. Bayonet probes were added to the sides of the sample kit as alternate mounting points. Additional locking features were added to the end effector and the filter stack to prevent inadvertent release during use. The opening mechanism was changed from one where the end effector was rocked laterally to defeat a ball detent to a twist-to-open threaded closure for similar reasons. Demonstrating, this sampling caddy’s effectiveness during a US EVA will allow us to address knowledge gaps identified in COSPAR reports and begin to define planetary protection requirements for life support systems on crewed missions to mars. This kit could also be used to collect contamination control samples during Artemis missions to verify requirements and could be easily modified for robotic sample collection.

Planetary Protection↗

ISS External Microorganisms: Collecting Planetary Protection Samples During Extravehicular Activity

We have developed, tested, and flown a caddy capable of collecting aseptic samples from external surfaces of the ISS (International Space Station). The sampling caddy is certified for use during US EVA (extra vehicular activity) and was launched to ISS in the summer of 2023. We are scheduled to collect samples from 6 locations outside ISS during an EVA in May of 2024. We will freeze these samples at -80°C on orbit and return them to Earth. We will then extract and sequence any DNA collected during the EVA using next generation sequencing technologies to characterize the community composition and function of each sample. Measuring the type and quantity of microbes present on the exterior of ISS will allow us to address knowledge gap 2B, “Acceptable levels of microbial/organic releases from humans and support systems” described in a 2019 COSPAR report. Collecting data about microbial release from current crewed vehicles will inform requirements for acceptable leak rates for future crewed missions to Mars. The sampling kit consists of eight commercially available, sterile, DNA free, macrofoam swabs ( 23 mm. diameter ) installed in custom aluminum end effectors. Each end effector is housed in and individual aluminum canister. Each canister contains a 0.2 μm Teflon filter to allow the interior volume to accommodate pressure changes without permitting microbial contaminants to enter the sterile interior volume. A handle repurposed from the space shuttle tile repair kit is used to remove the end effector from the sample canister, collect a sample by swabbing a surface and then replace the end effector in its canister. The canisters and end effectors were cleaned and assembled on Earth. Prior to installing the sterile swab the canisters and end effectors were sterilized in an autoclave at 134°C, 215 kPA, for 7 min.. The final assembly occurred in a sterilized class II biosafety cabinet. We will collect six samples from the 1) airlock vestibule, 2) airlock thermal cover, 3) a gap in the micrometeorite shielding near the airlock, 4) a handrail near the airlock, 5) the CDRA (Carbon Dioxide Removal Assembly) vent, and 6) the VES (Vacuum Exhaust System) vent. The remaining two swabs will be reserved as controls. One swab will be exposed during the EVA without touching any surfaces to act as a blank. The final swab will remain sealed until the entire sampling kit is returned to earth. Based on previously published results from the Russian segment, we hypothesize that there will be detectable microbes at some or all of these locations. Ground-based testing of this sampling caddy confirms that the swabs remain sterile as the canisters transition in and out of vacuum. We were able to retrieve, viable bacterial and fungal cells as well as DNA from samples collected from US space suits during vacuum chamber tests lasting as long as seven hours. Based on these results and feedback from the test subjects the sampling caddy was modified to improve ergonomics and meet US EVA safety requirements. Bayonet probes were added to the sides of the sample kit as alternate mounting points. Additional locking features were added to the end effector and the filter stack to prevent inadvertent release during use. The opening mechanism was changed from one where the end effector was rocked laterally to defeat a ball detent to a twist-to-open threaded closure for similar reasons. Demonstrating, this sampling caddy’s effectiveness during a US EVA will allow us to address knowledge gaps identified in COSPAR reports and begin to define planetary protection requirements for life support systems on crewed missions to mars. This kit could also be used to collect contamination control samples during Artemis missions to verify requirements and could be easily modified for robotic sample collection.

Planetary Protection↗