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Carrier-assisted One-pot Sample Preparation for Targeted Proteomics Analysis of Small Numbers of Human Cells

Protein analysis of small numbers of human cells is primarily achieved by targeted proteomics with antibody-based immunoassays, which have inherent limitations (e.g., low multiplex and unavailability of antibodies for new proteins). Mass spectrometry (MS)-based targeted proteomics has emerged as an alternative because it is antibody-free, high multiplex, and has high specificity and quantitation accuracy. Recent advances in MS instrumentation make MS-based targeted proteomics possible for multiplexed quantification of highly abundant proteins in single cells. However, there is a technical challenge for effective processing of single cells with minimal sample loss for MS analysis. To address this issue, we have recently developed a convenient protein carrier-assisted one-pot sample preparation coupled with liquid chromatography (LC) - selected reaction monitoring (SRM) termed cLC-SRM for targeted proteomics analysis of small numbers of human cells. This method capitalizes on using the combined excessive exogenous protein as a carrier and low-volume one-pot processing to greatly reduce surface adsorption losses as well as high-specificity LC-SRM to effectively address the increased dynamic concentration range due to the addition of exogeneous carrier protein. Its utility has been demonstrated by accurate quantification of most moderately abundant proteins in small numbers of cells (e.g., 10-100 cells) and highly abundant proteins in single cells. The easy-to-implement features and no need for specific devices make this method readily accessible to most proteomics laboratories. In this study, we have provided a detailed protocol for cLC-SRM analysis of small numbers of human cells including cell sorting, cell lysis and digestion, LC-SRM analysis, and data analysis. Further improvements in detection sensitivity and sample throughput are needed towards targeted single-cell proteomics analysis. We anticipate that cLC-SRM will be broadly applied to biomedical research and systems biology with the potential of facilitating precision medicine.

60 APPLIED LIFE SCIENCES↗

Lightning Characteristics Associated With Storm Modes Observed During RELAMPAGO

Global satellite studies show a maximum in deep convection and lightning downstream of the Andes in subtropical South America. The Remote sensing of Electrification, Lightning, And Mesoscale/microscale Processes with Adaptive Ground Observations (RELAMPAGO) field campaign was designed to investigate the physical processes that contribute to the rapid development of deep convection and mesoscale convective systems (MCSs) in Argentina. A lightning mapping array (LMA) was deployed to Argentina as part of RELAMPAGO to collect lightning observations from extreme storms in the region. This study combines lightning data from the LMA and the Geostationary Lightning Mapper onboard GOES-16 with 1-km gridded radar data to examine the electrical characteristics of a variety of convective storms throughout their life cycle observed during RELAMPAGO. Results from the full campaign show 48% of flashes are associated with deep convection that occurs along the eastern edge of the Sierras de Córdoba (SDC) overnight. These flashes are 65 km 2 smaller on average compared to stratiform flashes, which occur most frequently 50–100 km east of the SDC in the early morning hours, consistent with the upscale growth of MCSs off the terrain. Analysis of the 13–14 December MCS shows that sharp increases in flash rates correspond to deep and wide convective cores that have high graupel and hail mass, 35-dBZ volume, and ice water path. This work validates previous satellite studies of lightning in the region, but also provides higher spatial and temporal resolution information across the convective life cycle that has not been available in previous studies.

54 ENVIRONMENTAL SCIENCES↗

Lighting System Control Data to Improve Design and Operation: Tunable Lighting System Data from NICU Patient Rooms

The advancement of LED and controls technology, computing capacity, and software provides new opportunities for researchers and designers to work together to further optimize spaces for occupant benefit. Here, lighting system control data from five neonatal intensive care unit patient rooms was collected over a 25-week monitoring period and analyzed to better understand occupant response to a tunable lighting system with automatic transitions throughout the day. Lighting systems are very rarely refined after installation based on actual use. Objective data detailing how the lighting system is used by the actual occupants highlights the opportunities for optimization after installation and provides insight for improving the next design. As use of the data becomes more commonplace, it can be leveraged for design recommendations. The collection of the data required no additional cost beyond the time for examining the data. The analysis revealed several clear opportunities for improvement, including adjustments to the default control setting at night, re-labeling of the control stations, and adjustments to the nighttime fade rate. The patient room occupants were active users of the different zones, dimming options, and manual overrides made available by the lighting system.

60 APPLIED LIFE SCIENCES↗

Thiourea-Derived Chelating Ligands and Their Organometallic Compounds: Investigations into Their Anticancer Activity

Thiones have been investigated as ligands in metal complexes with catalytic and biological activity. We report the synthesis, characterization, and biological evaluation of a series of MII/III complexes of the general formulae [M II (cym)(L)Cl]X (cym = η 6 -p-cymene) or [M III (Cp*)(L)Cl]X (Cp* = η 5 -pentamethylcyclopentadienyl), where X = Cl - or PF 6 - , and L represents heterocyclic derivatives of thiourea. The thiones feature a benzyl-triazolyl pendant and they act as bidentate ligands via N,S-coordination to the metal centers. Several derivatives have been investigated by single-crystal X-ray diffraction analysis. NMR investigations showed a counterion-dependent shift of several protons due to the interaction with the counterions. These NMR investigations were complemented with X-ray diffraction analysis data and the effects of different counterions on the secondary coordination sphere were also investigated by DFT calculations. In biological studies, the Ir benzimidazole derivative was found to accumulate in the cytoplasm and it was the most cytotoxic derivative investigated.

60 APPLIED LIFE SCIENCES↗

Summary Status of the Space Acceleration Measurement System (SAMS), September 1993

The Space Acceleration Measurement System (SAMS) was developed to measure the microgravity acceleration environment to which NASA science payloads are exposed during microgravity science missions on the shuttle. Six flight units have been fabricated to date. The inaugural flight of a SAMS unit was on STS-40 in June 1991 as part of the First Spacelab Life Sciences mission. Since that time, SAMS has flown on six additional missions and gathered eighteen gigabytes of data representing sixty-eight days of microgravity environment. The SAMS units have been flown in the shuttle middeck and cargo bay, in the Spacelab module, and in the Spacehab module. This paper summarizes the missions and experiments which SAMS has supported. The quantity of data and the utilization of the SAMS data is described. Future activities are briefly described for the SAMS project and the Microgravity Measurement and Analysis project (MMAP) to support science experiments and scientists with microgravity environment measurement and analysis.

DeLombard, Richard↗

Boulders on Ceres

Introduction: In December 2015 the Dawn spacecraft moved into the Low Altitude Mapping Orbit (LAMO) around Ceres, encircling the dwarf planet at a distance of 400 km to the surface below. At this altitude, images of the on-board framing camera have a resolution of 36 meters per pixel, high enough to distinguish large boulders on the surface. Indeed, LAMO images show a multitude of boulders around what seem to be fresh craters. The average life-time of boulders on Dawn's previous target, Vesta, was estimated to be similar to that of Lunar boulders, as may be expected from the basaltic surface composition. The bulk composition of Ceres may be carbonaceous chondrite-like with significant contributions of clays, salt, and water ice. As such, the abundance and distribution of boulders on Ceres may be different from that on Vesta. We mapped, counted, and measured the diameter of boulders over the entire surface of Ceres. Our analysis of the data in combination with crater age estimates may provide clues to the physical nature and composition of the surface.

Ceres↗

Inland and Near-Shore Water Profiles Derived from the High-Altitude Multiple Altimeter Beam Experimental Lidar (MABEL)

The Advanced Topographic Laser Altimeter System (ATLAS) on the Ice, Cloud, and Land Elevation Satellite (ICESat-2) mission is a six beam, low energy, high repetition rate, 532-nanometer laser transmitter with photon counting detectors. Although designed primarily for detecting height changes in ice caps, sea ice, and vegetation, the polar-orbiting satellite will observe global surface water during its designed three-year life span, including inland waterbodies, coasts, and open oceans. In preparation for the mission, an ICESat-2 prototype, the Multiple Altimeter Beam Experimental Lidar (MABEL), was built and flown on high-altitude aircraft experiments over a range of inland and near-shore targets. The purpose was to test the ATLAS concept and to provide a database for developing an algorithm that detects along track surface water height and light penetration under a range of atmospheric and water conditions. The current analysis examines the data sets of three MABEL transects observed from 20 kilometers above ground of coastal and inland waters conducted in 2012 and 2013. Transects ranged from about 2 to 12 kilometers in length and included the middle Chesapeake Bay, the near-shore Atlantic coast at Virginia Beach, and Lake Mead. Results indicate MABEL's high capability for retrieving surface water height statistics with a mean height precision ofapproximately 5-7 centimeters per 100-meter segment length. Profiles of attenuated subsurface backscatter, characterized using a Signal to Background Ratio written in Log10 base, or LSBR (sub 0), were observed over a range of 1.3 to 9.3 meters, depending on water clarity and atmospheric background. Results indicate that observable penetration depth, although primarily dependent on water properties, was greatest when the solar background rate was low. Near-shore bottom reflectance was detected only at the Lake Mead site down to a maximum of 10 meters under a clear night sky and low turbidity of approximately 1.6 Nephelometric Turbidity Units (NTU). The overall results suggest that the feasibility of retrieving operational surface water height statistics from space-based photon counting systems such as ATLAS is very high for resolutions down to about 100 meters, even in partly cloudy conditions. The capability to observe subsurface backscatterprofiles is achievable but requires much longer transects of several hundreds of meters.

inland↗

Maximizing Spaceflight Biological Data with Omics Analytics: The NASA GeneLab Database

NASA’s GeneLab includes an open-access repository of some 250+ omics datasets generated by biological experiments relevant to spaceflight including simulated cosmic radiation and microgravity. In order to maximize the intelligibility of these data, particularly for users with limited bioinformatics background, GeneLab has become a knowledgebase platform converting raw genetic and proteomic signatures found in flight samples into biological and physiological meanings. A large community of more than 100 scientists has rallied behind GeneLab and organized into four Analysis Working Groups (AWGs: Animal, Plant, Microbe, and Multi-Omics). Together, the AWGs have gained scientific recognition worldwide by establishing a consortium in charge of adopting new complex standards for data analysis workflows and omics sample processing in a rapidly evolving field. We will demonstrate the usage of the repository with smart search capability, an online controlled-access toolshed "Galaxy" to process user data with vetted standard workflows, a workspace for data sharing and a data submission portal with ontology control for better metadata curation. The GeneLab visualization portal will also be demonstrated, showing how anyone without formal training in bioinformatics can now browse the space biology omics data to discover new biology and potential solutions to improve life in space.

Sylvain Vincent Costes↗

GeneLab: The NASA System Biology Platform for Space Omics Repository, Analysis and Visualization

NASA’s GeneLab includes an open-access repository of some 250+ omics datasets generated by biological experiments relevant to spaceflight including simulated cosmic radiation and microgravity. In order to maximize the intelligibility of these data, particularly for users with limited bioinformatics background, GeneLab has become a knowledgebase platform converting raw genetic and proteomic signatures found in flight samples into biological and physiological meanings. A large community of more than 100 scientists has rallied behind GeneLab and organized into four Analysis Working Groups (AWGs: Animal, Plant, Microbe, and Multi-Omics). Together, the AWGs have gained scientific recognition worldwide by establishing a consortium in charge of adopting new complex standards for data analysis workflows and omics sample processing in a rapidly evolving field. We will demonstrate the usage of the repository with smart search capability, an online controlled-access toolshed "Galaxy" to process user data with vetted standard workflows, a workspace for data sharing and a data submission portal with ontology control for better metadata curation. The GeneLab visualization portal will also be demonstrated, showing how anyone without formal training in bioinformatics can now browse the space biology omics data to discover new biology and potential solutions to improve life in space.

GeneLab↗

Future Model-Based Systems Engineering Vision and Strategy Bridge for NASA

A vision for the future of model-based systems engineering (MBSE) at NASA in 2029 and a strategy bridge towards that future are presented. Strategic thinking and leading change concepts were used to analyze reports and presentations on global trends and visionary thinking about the future of systems and digital engineering. The context, strategic time horizon, stakeholders, strategic challenges, strategic advantages, driving forces, and opportunities were considered. The analysis resulted in a future vision of MBSE that shows what NASA systems engineers and digital machines will do to perform rapid, extraordinary, and unprecedented missions. The NASA systems engineer, in this future vision, works with a global project team in a virtual and collaborative environment, engineers the system, and uses digital approaches as the routine and default way of working. The digital machines provide data-driven and automated mission designs; have a backbone of program and project management, systems engineering, and product life-cycle management; and are a knowledge-sharing infrastructure. The NASA systems engineer and the systems engineering team are envisioned to use digital machines to plan and perform rapid exploration missions, develop a digital twin that lasts across the life cycle, and develop enduring and adaptable systems. NASA has an engineering enterprise and a life-cycle management framework that endure, adapt, and respond. A strategy bridge based on the Baldrige Criteria for Performance Excellence Framework and lessons learned from a recent MBSE initiative illuminates a way forward from today to this desired future. The bridge lays out a strategy for leaders and recommends investments of today for immediate benefits and for benefits in 2029.

model-based systems engineering, digital engineeri↗

A hybrid method to evaluate the life cycle climate performance of heat pumps

Heat pumps have a significant impact on the climate, and numerous studies exist discussing the environmental impact of heat pumps quantitatively. Life Cycle Climate Performance (LCCP) is a widely accepted metric and can evaluate the carbon footprint of heat pumps from cradle to grave. LCCP calculation typically requires inputs like annual energy consumption, the material used, refrigerant type, and charge level, which are usually the intermediate or final results from heat pump simulation tools. Thus, a platform linking the heat pump simulation software, local weather conditions, and LCCP evaluation tool can simplify the heat pump design process, especially for complicated systems. This paper presents an LCCP evaluation tool accessing the outputs from the DOE Heat Pump Design Model and the building loads and temperature bin data from AHRI standard 210/240. As for case studies, this paper investigates the LCCP values and life cycle electricity costs (LCEC) of a given cooling-based heat pump with R410A in five typical US cities. In conclusion, this cooling-based heat pump has the highest LCCP value at Chicago and lowest LCCP value at Seattle. In contrast, the heat pump has the highest LCEC at Seattle and the lowest LCEC at Miami. As a future work, a more comprehensive carbon emission and cost analysis of heat pump systems will be conducted through all major cities around the world.

Shen, Bo↗

Underwater Noise Measurements around a Tidal Turbine in a Busy Port Setting

Acoustic emissions from current energy converters remain an environmental concern for regulators because of their potential effects on marine life and uncertainties about their effects stemming from a lack of sufficient observational data. Several recent opportunities to characterize tidal turbine sound emissions have begun to fill knowledge gaps and provide a context for future device deployments. In July 2021, a commercial-off-the-shelf hydrophone was deployed in a free-drifting configuration to measure underwater acoustic emissions and characterize a 25 kW-rated tidal turbine at the University of New Hampshire’s Living Bridge Project in Portsmouth, New Hampshire. Sampling methods and analysis were performed in alignment with the recently published IEC 62600-40 Technical Specification for acoustic characterization of marine energy converters. Results from this study indicate acoustic emissions from the turbine were below ambient sound levels and therefore did not have a significant impact on the underwater noise levels of the project site. As a component of Pacific Northwest National Laboratory’s Triton Field Trials (TFiT) described in this Special Issue, this effort provides a valuable use case for the IEC 62600-40 Technical Specification framework and further recommendations for cost-effective technologies and methods for measuring underwater noise at future current energy converter project sites.

tidal turbine↗

Development of the CELSS emulator at NASA. Johnson Space Center

The Closed Ecological Life Support System (CELSS) Emulator is under development. It will be used to investigate computer simulations of integrated CELSS operations involving humans, plants, and process machinery. Described here is Version 1.0 of the CELSS Emulator that was initiated in 1988 on the Johnson Space Center (JSC) Multi Purpose Applications Console Test Bed as the simulation framework. The run model of the simulation system now contains a CELSS model called BLSS. The CELSS simulator empowers us to generate model data sets, store libraries of results for further analysis, and also display plots of model variables as a function of time. The progress of the project is presented with sample test runs and simulation display pages.

Cullingford, Hatice S.↗

An expert system/ion trap mass spectrometry approach for life support systems monitoring

Efforts to develop sensor and control system technology to monitor air quality for life support have resulted in the development and preliminary testing of a concept based on expert systems and ion trap mass spectrometry (ITMS). An ITMS instrument provides the capability to identify and quantitate a large number of suspected contaminants at trace levels through the use of a variety of multidimensional experiments. An expert system provides specialized knowledge for control, analysis, and decision making. The system is intended for real-time, on-line, autonomous monitoring of air quality. The key characteristics of the system, performance data and analytical capabilities of the ITMS instrument, the design and operation of the expert system, and results from preliminary testing of the system for trace contaminant monitoring are described.

Palmer, Peter T.↗

Assessing Layered Materials in Gale Crater

The recent analysis of high resolution Mars Orbiter Camera (MOC) images of layered outcrops in equatorial regions reinforces two important ideas, which will probably eventually become paradigms, about Mars: 1) It has had a long, complex geologic history marked by change, as manifested in the different layers observed, and 2) Standing bodies of water existed for substantial lengths of time, indicating clement conditions possibly conducive to life. Although observations of layering and evidence for lakes and oceans has been reported for years based on Mariner 9 and Viking data, the MOC data show that this layering is much more pervasive and complex than previously thought. These layered sites are ideal for studying the geologic, and possibly biologic, history of Mars. Here, a layered site within Gale Crater is advocated as a Mars Exploration Rover (MER) target. This is one of the few layered areas within closed depressions (e.g., other craters and Vallis Marineris) that meets the landing site constraints and is accessible to both MER A and B.

Bridges, N. T.↗

Design and Verification

As future space missions become longer, an important aspect to consider is the habitability of the spacecraft. The amount of habitable volume affects not only astronaut comfort, but safety and mission success as well. However, as the volume is increased to aid in task performance, the weight of the vehicle and cost of the mission escalates in proportion. Pressure to reduce mission cost is constant, but the risk to mission success and crew survival must remain the priorities. The Constellation Program's Altair Lunar Lander is designed for short duration surface operation missions of seven to ten days. For short duration missions, humans will tolerate fairly primitive environmental situations provided the basic physiological arrangements are acceptable. However, for long-duration lunar surface operations, the living and operational spaces within which the crew work must provide both the essentials of life, as well as the support necessary for the crew to be productive in accomplishing their mission. The Altair is still in the preliminary design phase, which is the optimal time for Human Factors data to be provided to designers and engineers. A Human Centered Design (HCD) approach is being taken with our Human Factors evaluations. Human-in-the-loop testing is conducted using low-medium fidelity mock-ups of proposed lunar architecture. Based on current ConOps (Concept of Operations) procedures, a task analysis is performed in which individual tasks are combined into larger operational scenarios. Subjective and objective performance measures are gathered at both the task and scenario level. These scores are used to determine the functionality of the vehicle in terms of task performance. Results from these evaluations will highlight areas for design or operational improvement.

Thompson, Shelby G.↗

SPENVIS Implementation of End-of-Life Solar Cell Calculations Using the Displacement Damage Dose Methodology

This paper presents a method for using the SPENVIS on-line computational suite to implement the displacement damage dose (D(sub d)) methodology for calculating end-of-life (EOL) solar cell performance for a specific space mission. This paper builds on our previous work that has validated the D(sub d) methodology against both measured space data [1,2] and calculations performed using the equivalent fluence methodology developed by NASA JPL [3]. For several years, the space solar community has considered general implementation of the D(sub d) method, but no computer program exists to enable this implementation. In a collaborative effort, NRL, NASA and OAI have produced the Solar Array Verification and Analysis Tool (SAVANT) under NASA funding, but this program has not progressed beyond the beta-stage [4]. The SPENVIS suite with the Multi Layered Shielding Simulation Software (MULASSIS) contains all of the necessary components to implement the Dd methodology in a format complementary to that of SAVANT [5]. NRL is currently working with ESA and BIRA to include the Dd method of solar cell EOL calculations as an integral part of SPENVIS. This paper describes how this can be accomplished.

Walters, Robert↗

Advanced Stirling Convertor Testing at NASA Glenn Research Center

The NASA Glenn Research Center (GRC) has been testing high-efficiency free-piston Stirling convertors for potential use in radioisotope power systems (RPSs) since 1999. The current effort is in support of the Advanced Stirling Radioisotope Generator (ASRG), which is being developed by the U.S. Department of Energy (DOE), Lockheed Martin Space Systems Company (LMSSC), Sunpower, Inc., and the NASA GRC. This generator would use two high-efficiency Advanced Stirling Convertors (ASCs) to convert thermal energy from a radioisotope heat source into electricity. As reliability is paramount to a RPS capable of providing spacecraft power for potential multi-year missions, GRC provides direct technology support to the ASRG flight project in the areas of reliability, convertor and generator testing, high-temperature materials, structures, modeling and analysis, organics, structural dynamics, electromagnetic interference (EMI), and permanent magnets to reduce risk and enhance reliability of the convertor as this technology transitions toward flight status. Convertor and generator testing is carried out in short- and long-duration tests designed to characterize convertor performance when subjected to environments intended to simulate launch and space conditions. Long duration testing is intended to baseline performance and observe any performance degradation over the life of the test. Testing involves developing support hardware that enables 24/7 unattended operation and data collection. GRC currently has 14 Stirling convertors under unattended extended operation testing, including two operating in the ASRG Engineering Unit (ASRG-EU). Test data and high-temperature support hardware are discussed for ongoing and future ASC tests with emphasis on the ASC-E and ASC-E2.

Wilson, Scott D.↗