Search NASA⌕ Search

SEARCH · Search NASA

Results for “Cell viability”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 253 records · Page 14

A cryptic oxidoreductase safeguards oxidative protein folding in Corynebacterium diphtheriae

In many gram-positive Actinobacteria, including Actinomyces oris and Corynebacterium matruchotii , the conserved thiol-disulfide oxidoreductase MdbA that catalyzes oxidative folding of exported proteins is essential for bacterial viability by an unidentified mechanism. Intriguingly, in Corynebacterium diphtheriae , the deletion of mdbA blocks cell growth only at 37 °C but not at 30 °C, suggesting the presence of alternative oxidoreductase enzyme(s). By isolating spontaneous thermotolerant revertants of the mdbA mutant at 37 °C, we obtained genetic suppressors, all mapped to a single T-to-G mutation within the promoter region of tsdA , causing its elevated expression. Strikingly, increased expression of tsdA —via suppressor mutations or a constitutive promoter—rescues the pilus assembly and toxin production defects of this mutant, hence compensating for the loss of mdbA . Structural, genetic, and biochemical analyses demonstrated TsdA is a membrane-tethered thiol-disulfide oxidoreductase with a conserved CxxC motif that can substitute for MdbA in mediating oxidative folding of pilin and toxin substrates. Together with our observation that tsdA expression is upregulated at nonpermissive temperature (40 °C) in wild-type cells, we posit that TsdA has evolved as a compensatory thiol-disulfide oxidoreductase that safeguards oxidative protein folding in C. diphtheriae against thermal stress.

59 BASIC BIOLOGICAL SCIENCES↗

Effect of solar irradiation on extracellular enzymes of Aeromonas proteolytica

The bacterium Aeromonas proteolytica was selected for studying the effects of solar irradiation on extracellular enzymes because it produces an endopeptidase that is capable of degrading proteins and a hemolysin that is active in lysing human erythrocytes. Possible alterations in the rate of enzyme production in response to the test conditions are currently underway and are not available for this preliminary report. Completed viability studies are indicative that little difference exists among the survival curves derived for cells exposed to various components of ultraviolet irradiation in space.

Foster, B. G.↗

Self-consistent calculations and design considerations for a GaAs nipi doping superlattice solar cell

The authors present design constraints which show that a previously proposed GaAs nipi doping superlattice solar cell structure would not work as an efficient space solar cell. A structure based on the CLEFT process, which shows promise of being an efficient cell with very high radiation tolerance, is proposed. In order to test theoretically its viability and to optimize its design, self-consistent quantum mechanical calculations were made for a number of thicknesses of the n, i, and p layers and the dopings in the n and p layers. These results show that: 1) an i layer is not necessary; in fact, its presence makes it difficult to satisfy one of the key constraints; 2) a near-optimum design with 750-A thick n and p layers with dopings of 2.5E18/cu cm and a selective contact separation of 20 microns would yield both high efficiency and very high radiation tolerance.

Clark, Ralph O.↗

Spaceflight-induced Changes in Microbial Virulence and the Impact to the Host Immune Response

INTRODUCTION Over the past 50 years, many microorganisms have displayed unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in stress resistance, biofilm production, antibiotic sensitivity, final cell concentration, gene expression, enhanced host-pathogen interaction, and virulence. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system; consisting of altered leukocyte distribution, reductions in T and NK cell function, altered cytokine profiles, and reactivation of latent herpesviruses. Further, astronauts have some degree of clinical incidence, primarily infectious disease episodes and atopic dermatitis. The impact of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. SPECIFIC AIMS In this study, the following Specific Aims are being investigated, using the microbial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli (EHEC). Aim 1: Characterize the effect of spaceflight analogue culture on microbial pathogenesis-related stress responses and in vitro host-pathogen interactions. Analyses include microbial stress responses as well as colonization and viability following pathogen challenge of three-dimensional (3-D) tissue co-culture models containing immune cells. We specifically selected obligate and opportunistic pathogens that are medically important and have been or are likely to be found aboard spacecraft. Aim 2: Characterize the effect of spaceflight analogue culture on the virulence potential of pathogenic microorganisms. Virulence will be assessed using a mouse model of infection. SIGNIFICANCE The goal of this study is to use spaceflight analogue conditions to gain insight into the breadth of medically-significant International Space Station microorganisms that have altered virulence and the impact of those changes on the immune response of the host. This information will provide critical understanding into the impact of microgravity on potential alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.

C M Ott↗

SPACEFLIGHT-INDUCED CHANGES IN MICROBIAL VIRULENCE AND THE IMPACT TO THE HOST IMMUNE RESPONSE

INTRODUCTION Over the past 50 years, microorganisms have displayed unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, and virulence. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system; consisting of altered leukocyte distribution, reductions in T and NK cell function, altered cytokine profiles, and reactivation of latent herpesviruses. Further, astronauts have some degree of clinical incidence, primarily infectious disease episodes and atopic dermatitis. The impact of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. SPECIFIC AIMS In the Host-Microorganism, Microbial Response aspect of this study, the following Specific Aims are being investigated, using the microbial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. Aim 1: Characterize the effect of spaceflight analogue culture on microbial pathogenesis-related stress responses and in vitro host-pathogen interactions. Analysis will include microbial stress responses as well as colonization and viability following pathogen challenge of three-dimensional (3-D) tissue co-culture models containing immune cells. We specifically selected obligate and opportunistic pathogens that are medically important and have been or are likely to be found aboard spacecraft. Aim 2: Characterize the effect of spaceflight analogue culture on the virulence potential of pathogenic microorganisms. Changes in virulence will be assessed using a mouse model of infection. SIGNIFICANCE The goal of this study is to use spaceflight analogue conditions to gain insight into the breadth of medically-significant International Space Station microorganisms that have altered virulence and the impact of those changes on the immune response of the host, including astronaut immune cells. This information will provide critical understanding into the impact of microgravity on potential alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.

C. M. Ott↗

Long-life LiNi 0.5 Mn 1.5 O 4 /graphite lithium-ion cells with an artificial graphite-electrolyte interface

The high operating voltage of a spinel LiNi 0.5 Mn 1.5 O 4 (LNMO) cathode leads to electrolyte decomposition and accelerated deterioration of the electrode/electrolyte interface in lithium-ion cells. Aggressive side reactions prevent long-term cycling and hinder its practical application. Advanced characterization shows that the high-voltage operation makes the graphite anode react with the electrolyte, particularly with the lithium salt in the electrolyte, resulting in massive active lithium inventory loss and capacity fade. The side reactions at the graphite are identified as the main culprit for the LNMO/graphite full cell decay. Here, we demonstrate that an artificial solid-electrolyte interface layer consisting of an ultra-thin Al 2 O 3 film, generated via atomic layer deposition (ALD) at the graphite surface, can successfully isolate the graphite from the electrolyte and inhibit the undesired reactions. Consequently, the capacity and cycling stability of the LNMO/graphite cell are drastically improved. After 300 cycles, the capacity retention of LNMO/graphite cell increases from 57.7% to 98.6% with the ALD-coated graphite. Furthermore, our findings provide valuable insights into the LNMO/graphite cell degradation and suggest a simple and robust, yet highly effective, approach for the practical viability of high-voltage lithium-ion batteries.

25 ENERGY STORAGE↗

Evaluation of the recombinant proteins RlpB and VacJ as a vaccine for protection against Glaesserella parasuis in pigs

Abstract Background Glaesserella parasuis , the causative agent of Glӓsser’s disease, is widespread in swine globally resulting in significant economic losses to the swine industry. Prevention of Glӓsser’s disease in pigs has been plagued with an inability to design broadly protective vaccines, as many bacterin based platforms generate serovar or strain specific immunity. Subunit vaccines are of interest to provide protective immunity to multiple strains of G. parasuis . Selected proteins for subunit vaccination should be widespread, highly conserved, and surface exposed. Results Two candidate proteins for subunit vaccination (RlpB and VacJ) against G. parasuis were identified using random mutagenesis and an in vitro organ culture system. Pigs were vaccinated with recombinant RlpB and VacJ, outer membrane proteins with important contributions to cellular function and viability. Though high antibody titers to the recombinant proteins and increased interferon-γ producing cells were found in subunit vaccinated animals, the pigs were not protected from developing systemic disease. Conclusions It appears there may be insufficient RlpB and VacJ exposed on the bacterial surface for antibody to bind, preventing high RlpB and VacJ specific antibody titers from protecting animals from G. parasuis . Additionally, this work confirms the importance of utilizing the natural host species when assessing the efficacy of vaccine candidates.

Hau, Samantha J.↗

Angle of Incidence Characterization of Six Laminated Solar Cells for 2020 DTU Fotonik Inter-Laboratory Comparison Study

Photovoltaic energy prediction models include functions or modifiers to account for sun angle reflection losses. These functions may be known interchangeably as Angle of Incidence (AOI) or Incident Angle Modifier (IAM). While standards exist, there is no universally accepted single best practice for developing these functions. They can be generated through characterization of representative modules or single cells, in natural sunlight or indoors using simulated light sources. Repeatability of measurements and the viability of cross-laboratory comparisons are critical to confidence in validation of both methods. To investigate the differences between methods and labs, The Technical University of Denmark (DTU) initiated an international round-robin test comparison between several key test labs with AOI measurement capability. A total of six minimodules were provided in three different cell/interconnect/backsheet combinations. Sandia characterized these minimodules using methods developed over two decades specifically for the outdoor characterization of full-size photovoltaic modules. This report documents the characterization results, summarizes key observations and tabulates the processed data for comparison to results provided by other characterization labs.

14 SOLAR ENERGY↗

CT584 Is Not a Protective Vaccine Antigen against Respiratory Chlamydial Challenge in Mice

Background: Chlamydia trachomatis is the most prevalent bacterial sexually transmitted pathogen in humans worldwide. Since chlamydial infection is largely asymptomatic with the potential for serious complications, a preventative vaccine is likely the most viable long-term answer to this public health threat. Cell-free protein synthesis (CFPS) utilizes the cellular protein manufacturing machinery decoupled from the requirement for maintaining cellular viability, offering the potential for flexible, rapid, and decentralized production of recombinant protein vaccine antigens. Methods: Here, we use CFPS to produce the full-length putative chlamydial type three secretion system (T3SS) needle-tip protein, CT584, for evaluation as a vaccine antigen in mouse models. High-speed atomic force microscopy (HS-AFM) (RIBM, Tsukuba, Japan) imaging and computer simulations confirm that CFPS-produced CT584 retains a native-like structure prior to immunization. Female mice were primed with CT584 adjuvanted with CpG-1826 intranasally (i.n.) or CpG-1826 + Montanide ISA 720 intramuscularly (i.m.), followed four weeks later by an i.m. boost before respiratory challenge with 10 4 inclusion forming units (IFU) of Chlamydia muridarum. Results: Immunization with CT584 generated robust antibody responses but weak cell-mediated immunity and failed to protect against i.n. challenge as demonstrated by body weight loss, increased lung weights, and the presence of high numbers of IFUs in the lungs. Conclusion: While CT584 was not a protective vaccine candidate, the speed and flexibility with which CFPS can be used to produce other potential chlamydial antigens make it an attractive technique for antigen production.

60 APPLIED LIFE SCIENCES↗

Increasing the Thermal Stability of Aluminum Titanate for Solid Oxide Fuel Cell Anodes

Solid-oxide fuel cells (SOFCs) show great potential as a power source for future space exploration missions. Because SOFCs operate at temperatures significantly higher than other types of fuel cells, they can reach overall efficiencies of up to 60% and are able to utilize fossil fuels. The SOFC team at GRC is leading NASA's effort to develop a solid oxide fuel cell with a power density high enough to be used for aeronautics and space applications, which is approximately ten times higher than ground transport targets. layers must be able to operate as a single unit at temperatures upwards of 900'C for at least 40,000 hours with less than ten percent degradation. One key challenge to meeting this goal arises from the thermal expansion mismatch between different layers. The amount a material expands upon heating is expressed by its coefficient of thermal expansion (CTE). If the CTEs of adjacent layers are substantially different, thermal stresses will arise during the cell's fabrication and operation. These stresses, accompanied by thermal cycling, can fracture and destroy the cell. While this is not an issue at the electrolyte-cathode interface, it is a major concern at the electrolyte-anode interface, especially in high power anode-supported systems. electrolyte are nearly identical. Conventionally, this has been accomplished by varying the composition of the anode to match the CTE of the yittria-stabilized zirconia (YSZ) electrolyte (approx.10.8x10(exp -6/degC). A Ni/YSZ composite is typically used as a base material for the anode due to its excellent electrochemical properties, but its CTE is about 13.4x10(exp -6/degC). One potential way to lower the CTE of this anode is to add a small percentage of polycrystalline Al2TiO5, with a CTE of 0.68x10(exp -6/degC, to the Ni/YSZ base. However, Al2TiO5 is thermally unstable and loses its effectiveness as it decomposes to Al2O3 and TiO2 between 750 C and 1280 C. be used as additives to increase the thermal stability of Al2TiO5 in SOFC operating conditions without adversely affecting the electrochemical properties of the SOFC anode. Three candidate materials were chosen through an extensive literature review: MgO, Fe2O3, and ZrTiO4. Although all three have been shown to prevent Al2TiO5 decomposition under various conditions, their effectiveness in the temperature range and atmosphere of the SOFC has not yet been evaluated. Several batches of Al2TiO5 with varying amounts of additives were prepared, exposed to reducing and oxidizing atmospheres at elevated temperatures, and the resulting decomposition of Al2TiO5 was measured. The most promising additives were further evaluated with the goal of ultimately preparing low CTE anodes that are chemically compatible to current systems. Adding minor constituents to stabilize Al2TiO5 could ultimately preserve its low CTE for the life of the fuel cell and improve the cell's long-term performance without a drop in anode conductivity. Further, these low CTE filler additions could allow the use of new sulfur tolerant anode materials, improving the viability of SOFCs for future aeronautics and space applications. Every SOFC consists of a cathode and an anode separated by an electrolyte, These three One way to avoid this problem is to design the cell such that the CTEs of the anode and The objective of this summer research project was to evaluate several materials that could

Bender, Jeffrey B.↗

Peripheral blood hematology, plasma biochemistry, and the optimization of an in vitro immune-based assay in the brown watersnake ( Nerodia taxispilota )

Reptiles represent a phylogenetic lineage that provides a unique link between ectothermic anamniotes and endothermic amniotes. Compared to mammalian and avian species, our understanding of the reptilian immune system is greatly lacking. This gap in knowledge is largely due to an absence of established immune-based assays or specific reagents for these species. In the present study, brown watersnakes (Nerodia taxispilota) were live-captured in the wild, sexed, weighed, measured, bled via the caudal vein and released. At 24 hr post-collection, peripheral blood leukocytes were enriched and evaluated with an established mammalian in-vitro lymphocyte proliferation assay. Snake peripheral blood leukocyte enrichment yielded > 90% lymphocytes with viabilities averaging 81.5%. Baseline physiologic data for N. taxispilota, including hematology and total solids, leukocyte differentials, cell recovery and plasma biochemistry, were also collected. Cells cultured with Concanavalin A exhibited significantly increased proliferation at both 72 and 96 hr. Furthermore, these preliminary results show that enriched peripheral blood from wild-caught N. taxispilota provides a sufficient yield of leukocytes that can be cultured and functionally evaluated using a standard mammalian in-vitro immune-based assay.

59 BASIC BIOLOGICAL SCIENCES↗

Reduction of Thermal Residual Stresses in Advanced Metallic Composites Based upon a Compensating/Compliant Layer Concept

High residual stresses within metal and intermetallic matrix composite systems can develop upon cooling from the processing temperature to room temperature due to the coefficient of thermal expansion (CTE) mismatch between the fiber and matrix. As a result, within certain composite systems, radial, circumferential, and/or longitudinal cracks have been observed to form at the fiber-matrix interface region. The compliant layer concept (insertion of a compensating interface material between the fiber and matrix) has been proposed to reduce or eliminate the residual stress buildup during cooling, and thus minimize cracking. The present study investigates elastic-plastically the viability of the proposed compliant layer concept. A detailed parametric study was conducted utilizing a unit cell model consisting of three concentric cylinders to determine the required character (i.e., thickness and mechanical properties) of the compliant layer as well as its applicability. The unknown compliant layer mechanical properties were expressed as ratios of the corresponding temperature dependent Ti-24Al-11Nb (a/o) matrix properties. The fiber properties taken were those corresponding to SCS-6 (SiC). Results indicated that the compliant layer can be used to reduce, if not eliminate, radial and circumferential residual stresses within the fiber and matrix and therefore also reduce or eliminate the radial cracking. However, with this decrease in in-plane stresses, one obtains an increase in longitudinal stress, thus potentially initiating longitudinal cracking. Guidelines are given for the selection of a specific compensating/ compliant material, given a perfectly bonded system.

S. M. Arnold↗

Microgravity Effecs During Fertilization, Cell Division, Development, and Calcium Metabolism in Sea Urchins

Calcium loss and muscle atrophy are two of the main metabolic changes experienced by astronauts and crew members during exposure to microgravity in space. For long-term exposure to space it is crucial to understand the underlying mechanisms for altered physiological functions. Fundamental occurrences in cell biology which are likely to depend on gravity include cytoskeletal dynamics, chromatin and centrosome cycling, and ion immobilization. These events can be studied during fertilization and embryogenesis within invertebrate systems. We have chosen the sea urchin system to study the effects of microgravity on cytoskeletal processes and calcium metabolism during fertilization, cell division, development, and embryogenesis. Experiments during an aircraft parabolic flight (KC-135) demonstrated: (1) the viability of sea urchin eggs prior to fertilization, (2) the suitability of our specimen containment system, (3) the feasibility of fertilization in a reduced gravity environment (which was achieved during 25 seconds of reduced gravity under parabolic flight conditions). Two newly developed pieces of spaceflight hardware made further investigations possible on a spaceflight (STS-77); (1) the Aquatic Research Facility (ARF), and (2) the Fertilization Syringe Unit (FSU). The Canadian Space Agency developed ARF to conduct aquatic spaceflight experiments requiring controlled conditions of temperature, humidity, illumination, and fixation at predetermined time points. It contained a control centrifuge which simulated the 1 g environment of earth during spaceflight. The FSU was developed at the Kennedy Space Center (KSC) by the Bionetics Corporation specifically to enable the crew to perform sea urchin fertilization operations in space.

Schatten, Heide↗

Mitochondrial Oxidative Stress: Importance for Skeletal Structure and Responses to Simulated Spaceflight

Spaceflight environment poses challenges to the human body. Both microgravity and radiation may lead to excess production of reactive oxygen species (ROS) and resulting oxidative stress and tissue damage. Specifically in bone, elevated ROS can contribute to excess bone resorption by osteoclasts over bone formation by osteoblasts, reduced viability of osteocytes, and ultimately, osteoporosis. Thus, we hypothesized that suppression of mitochondrial ROS in bone cells improves overall bone structure in the adult skeleton and skeletal defenses from spaceflight. To begin to test our hypothesis, we (1) modified ROS levels in bone cells using mCAT (Malonyl CoA-acyl carrier protein transacylase) mice, which overexpress the human anti-oxidant catalase gene targeted to the mitochondria. We also increased ROS and stimulated skeletal remodeling by exposing mice to simulated spaceflight (hindlimb-unloading and total body-irradiation) or sham treatment. When challenged with treatment, bones from wildtype mice showed elevated levels of oxidative damage whereas mCAT mice did not. Treatment caused expected bone loss in wildtype mice. Treatment caused deficits in microarchitecture of mCAT mice, although lower in magnitude than wildtype. In conclusion, our results indicate mitochondrial ROS generation in osteoblast and osteoclast lineage cells of skeletal tissue contributes to skeletal remodeling and quenching oxidative damage may improve the skeletal responses to spaceflight.

Radiation↗

Electrochemically Engineered, Highly Energy-Efficient Conversion of Ethane to Ethylene and Hydrogen below 550 °C in a Protonic Ceramic Electrochemical Cell

Ethylene is one of the largest building blocks in the petrochemical industry, mainly produced by steam cracking of ethane derived from naphtha or shale gas at high temperatures (>800 °C). Despite its technical maturity and economic competitiveness, the thermal steam cracking of ethane is highly energy-intensive. In this work, an electrochemically engineered direct conversion process of ethane to produce hydrogen and ethylene using a planar protonic ceramic membrane reactor with a bi-functional three-dimensional catalytic electrode is reported, with a single-pass ethane conversion of 40% and ethylene yield of 26.7% at 550 °C. Compared with the industrial ethane steam cracking, this method saves process energy input by 45.1% and improves process energy efficiency by 50.6%, based on comprehensive process simulation using Aspen Plus software. Further, steam electrolysis treatment under the solid oxide electrolysis cell mode can regenerate the system’s catalytic performance and significantly alleviate catalytic degradation by 74%, demonstrating high techno-economic viability.

bi-functional electrode↗

Human mononuclear cell function after 4 degrees C storage during 1-G and microgravity conditions of spaceflight

Future space missions of long duration may require that autologous leukocytes be stored in flight for infusion to restore normal immune competence in crewmembers. Peripheral blood mononuclear cells (PBMNCs), as leukocyte concentrates in autologous plasma and 2% dextrose, were stored in the microgravity conditions provided by the U.S. Space Shuttle Columbia (Mission 61-C). Activity of PBMNC after space flight was compared with that from a series of preflight ground control experiments, which demonstrated in culture a progressive daily loss in mitogen-stimulated protein synthesis at 24 h and thymidine uptake at 72 h after storage for 7 d at 4 degrees C. Post-storage viabilities were at least 90% as determined by trypan dye exclusion. A progressive reduction in the percentage of PBMNC expressing cell-surface phenotype markers, which was similar for monocytes, B cells, and T-cell subsets, also occurred after storage. The ability of PBMNC, stored for 8 d in Columbia's middeck, to become activated and proliferate in vitro was similar to that of cells that remained in identical flight lockers on the ground as 1-G controls, thus indicating that PBMNCs were not adversely affected by storage under microgravity conditions.

NASA Discipline Number 00-00↗

Laser annealing of amorphous/poly: Silicon solar cell material flight experiment

The preliminary design proposed for the microelectronics materials processing equipment is presented. An overall mission profile, description of all processing steps, analysis methods and measurement techniques, data acquisition and storage, and a preview of the experimental hardware are included. The goal of the project is to investigate the viability of material processing of semiconductor microelectronics materials in a micro-gravity environment. The two key processes are examined: (1) Rapid Thermal Annealing (RTA) of semiconductor thin films and damaged solar cells, and (2) thin film deposition using a filament evaporator. The RTA process will be used to obtain higher quality crystalline properties from amorphous/poly-silicon films. RTA methods can also be used to repair radiation-damaged solar cells. On earth this technique is commonly used to anneal semiconductor films after ion-implantation. The damage to the crystal lattice is similar to the defects found in solar cells which have been exposed to high-energy particle bombardment.

Cole, Eric E.↗

A conceptual design of a large aperture microwave radiometer geostationary platform

A conceptual design of a Large Aperture Microwave Radiometer (LAMR) Platform has been developed and technology areas essential to the design and on-orbit viability of the platform have been defined. Those technologies that must be developed to the requirement stated here for the LAMR mission to be viable include: advanced radiation resistant solar cells, integrated complex structures, large segmented reflector panels, sub 3 kg/m(exp 2) areal density large antennas, and electric propulsion systems. Technology areas that require further development to enhance the capabilities of the LAMR platform (but are not essential for viability) include: electrical power storage, on-orbit assembly, and on-orbit systems checkout and correction.

Garn, Paul A.↗