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At least 739 records · Page 41

Evolution of safety-critical requirements post-launch

This paper reports the results of a small study of requirements changes to the onboard software of three spacecraft subsequent to launch. Only those requirement changes that resulted from post-launch anomalies (i.e., durring operations) were of interest here, since the goal was to better understand the relationship between critical anomolies during operations and how safety-critical requirements evolve.

requirements↗

Evapotranspiration Based Irrigation Trials Examine Water Requirement, Nitrogen Use, and Yield of Romaine Lettuce in the Salinas Valley

Cool season vegetables require adequate soil moisture to assure that maximum yield and quality are achieved. On California’s central coast, where the majority of cool season vegetables are produced in the US, long-term overpumping of irrigation water has reduced groundwater levels and led to environmental degradation. Two evapotranspiration (ET) based irrigation field trials were performed near Salinas CA (USA) to determine if ET-based irrigation scheduling could conserve water while producing romaine lettuce (cv. Sun Valley) of commercially viable yield. Sprinklers were used for seed germination and crop establishment. Four drip irrigation treatments were then imposed using a randomized complete block design with six replications. The CropManage decision support model was used to estimate the full (100%) crop water requirement based mainly on ET replacement. Other treatments included 50% 75% and 150% of the full water requirement. The 100% treatment received 185 mm of water in 2015 and 247 mm in 2016, both of which were well below prior guidance and grower reports. Yields from the 100% and 150% treatments were not significantly different and were similar to industry average, while yields were significantly lower for the 50% and 75% treatments. The 100% treatment had the highest water use efficiency, and the 100% and 150% treatments together had the highest nitrogen recovery efficiency. Irrigation of romaine near the 100% ET replacement level can potentially reduce environmental impacts associated with nitrate leaching and surface runoff.

CropManage decision support system↗

Modeling NASA’s Procedural Requirement Processes - Implications for Digital Future

The National Aeronautics and Space Administration (NASA) has an ongoing Digital Transformation effort and to leverage and showcase the power of Digital Transformation, an effort is underway to develop an integrated, datacentric, model representing NASA’s key process requirements. The task was divided into three phases: As Is modeling, Analysis, and To Be Planning. As part of this effort, a team has completed the first Phase I of the modeling task and is nearing completion of the second phase. This effort will capture the key elements as requirements, responsibilities, allocations, roles, products, and associated lifecycle elements. The scope of modeling included NASA’s NPR 7120.5 (Project and Program Management), NPR 7123.1 (Systems Engineering) and NPRs 8705.2 (Risk classification for Robotic Missions) and 8705.4 (Human-Rating Requirements for Space Missions). This paper will summarize the approach, scope, parsing patterns applied, metamodel, and associated workflows for the As-Is modeling. It will also summarize the results and insights gleaned during that phase, including the review process. These insights have informed the analysis and will be discussed. The analysis modeling phase will also be summarized including how the stakeholders were engaged, how the common elements were handled and dispositioned, and will also describe some of the plans for the future of NASA NPDs and NPRs.

Systems Engineering↗

Modeling NASA’s Procedural Requirement Processes – Implications for a Digital Future

The National Aeronautics and Space Administration (NASA) has an ongoing Digital Transformation effort and to leverage and showcase the power of Digital Transformation, an effort is underway to develop an integrated, datacentric, model representing NASA’s key process requirements. The task was divided into three phases: As Is modeling, Analysis, and To Be Planning. As part of this effort, a team has completed the first Phase I of the modeling task and is nearing completion of the second phase. This effort will capture the key elements as requirements, responsibilities, allocations, roles, products, and associated lifecycle elements. The scope of modeling included NASA’s NPR 7120.5 (Project and Program Management), NPR 7123.1 (Systems Engineering) and NPRs 8705.2 (Risk classification for Robotic Missions) and 8705.4 (Human-Rating Requirements for Space Missions).

NPR↗

Compartment-specific energy requirements of photosynthetic carbon metabolism in Camelina sativa leaves

Abstract Main conclusion The oxidative pentose phosphate pathway provides cytosolic NADPH yet reduces carbon and energy use efficiency. Repressing this pathway and introducing cytosolic NADPH-dependent malate dehydrogenase may increase crop yields by ≈5%. Abstract Detailed knowledge about plant energy metabolism may aid crop improvements. Using published estimates of flux through central carbon metabolism, we phenotype energy metabolism in illuminated Camelina sativa leaves (grown at 22 °C, 500 µmol photons m −2 s −1 ) and report several findings. First, the oxidative pentose phosphate pathway (OPPP) transfers 3.3% of the NADPH consumed in the Calvin–Benson cycle to the cytosol. NADPH supply proceeds at about 10% of the rate of net carbon assimilation. However, concomitantly respired CO 2 accounts for 4.8% of total rubisco activity. Hence, 4.8% of the flux through the Calvin–Benson cycle and photorespiration is spent on supplying cytosolic NADPH, a significant investment. Associated energy requirements exceed the energy output of the OPPP. Thus, autotrophic carbon metabolism is not simply optimised for flux into carbon sinks but sacrifices carbon and energy use efficiency to support cytosolic energy metabolism. To reduce these costs, we suggest bioengineering plants with a repressed cytosolic OPPP, and an inserted cytosolic NADPH-dependent malate dehydrogenase tuned to compensate for the loss in OPPP activity (if required). Second, sucrose cycling is a minor investment in overall leaf energy metabolism but a significant investment in cytosolic energy metabolism. Third, leaf energy balancing strictly requires oxidative phosphorylation, cofactor export from chloroplasts, and peroxisomal NADH import. Fourth, mitochondria are energetically self-sufficient. Fifth, carbon metabolism has an ATP/NADPH demand ratio of 1.52 which is met if ≤ 21.7% of whole electron flux is cyclic. Sixth, electron transport has a photon use efficiency of ≥ 62%. Last, we discuss interactions between the OPPP and the cytosolic oxidation–reduction cycle in supplying leaf cytosolic NADPH.

59 BASIC BIOLOGICAL SCIENCES↗

CLMcrop yields and water requirements: avoided impacts by choosing RCP 4.5 over 8.5

Here, we perform CLMcrop simulations of the 20th and 21st centuries to assess potential avoided impacts in (a) crop yield losses and (b) water demand increases if humanity were to choose the representative concentration pathway (RCP) 4.5 instead of 8.5. RCP 8.5 imposes more extreme climatic changes on CLMcrop, while simultaneously exposing the crops to higher CO2 fertilization than RCP 4.5. As a result CLMcrop simulates global to regional scale changes in yield and water requirements for RCP 8.5 that exceed and sometimes more than double the RCP 4.5 changes relative to today. Under RCP 4.5 then, human societies may confront easier adaptation to changes in crop yields and water requirements. Under both RCPs, CLMcrop projects declining global yields for C3 crops (e.g., wheat, soybean, rice) without CO 2 fertilization and C4 crops (corn, sugarcane) without irrigation. Yield declines of 3 t ha -1 stand out in parts of tropical and subtropical Africa and South America (presently areas of rapid agricultural expansion) and are due to increasing plant respiration and decreasing soil moisture, both due to rising temperatures. Irrigation and CO 2 fertilization mitigate yield losses and in some cases lead to gains, so irrigation may help maintain or increase current yields through the 21st century. However, simulated global irrigation requirements increase: as much as 23 % for C4 crops without CO 2 fertilization under RCP 8.5 and as little as 3 % for C4 crops with CO 2 fertilization under RCP4.5. Nitrogen fertilized crops display greater vulnerability to climate and environmental change than unfertilized crops in our simulations; still relative to unfertilized crops, they deliver significantly higher yields and remain indispensable in supporting a more populous and affluent humanity. These CLMcrop results broadly agree with previously published outcomes for the 21st century. We describe in this article a new version of CLMcrop that represents prognostic crop behavior not only in the mid-latitudes but also the tropics.

54 ENVIRONMENTAL SCIENCES↗

Reliable Thermoelectric Module Design under Opposing Requirements from Structural and Thermoelectric Considerations

Structural reliability of thermoelectric generation (TEG) systems still remains an issue especially for applications such as large scale industrial or automobile exhaust heat recovery where a TEG system is subject to dynamic loads and thermal cycling. Traditional thermoelectric (TE) system design and optimization techniques, focused on performance alone, could result in designs that may fail during operation as the geometric requirements for optimal performance (especially the power) are often in conflict with the requirements for mechanical reliability. This study focused on reducing the thermomechanical stresses in a TEG system without compromising the optimized system performance. Finite element simulations were carried out to study the effect of TE element (leg) geometry such as the leg length and cross-sectional shape under constrained material volume requirements. Results indicated that the element length has a major influence on the element stresses whereas the regular cross-sectional shapes have minor influence. Additionally, the impact of TE element stresses on the mechanical reliability is evaluated with the brittle material failure theory based on Weibull analysis. An alternate couple configuration that relies on the industry practice of redundant element design is investigated. Results showed that the alternate configuration considerably reduced the TE element and metallization stresses thereby enhancing the structural reliability with little trade-off in the optimized performance. The proposed alternate configuration could serve as a potential design modification for improving the reliability of systems optimized for thermoelectric performance.

36 MATERIALS SCIENCE↗

V-ATPase-dependent luminal acidification is required for endocytic recycling of a yeast cell wall stress sensor, Wsc1p

Highlights: •A targeted genome screen identified 5 gene groups affecting Wsc1p recycling. •V-ATPase-dependent luminal acidification is required for Wsc1p recycling. •Activity of V-ATPase might be required for cargo recognition by the retromer complex. -- Abstract: Wsc1p is a major cell wall sensor protein localized at the polarized cell surface. The localization of Wsc1p is maintained by endocytosis and recycling from endosomes back to the cell surface, but changes to the vacuole when cells are subjected to heat stress. Exploiting this unique property of Wsc1p, we screened for yeast single-gene deletion mutants exhibiting defects in Wsc1p trafficking. By expressing 3GFP-tagged Wsc1p in mutants with deleted genes whose function is related to intracellular trafficking, we identified 5 gene groups affecting Wsc1p trafficking, impaired respectively in endocytic internalization, multivesicular body sorting, the GARP complex, endosomal maturation/vacuolar fusion, and V-ATPase. Interestingly, deletion of the VPH1 gene, encoding the V{sub o} subunit of vacuolar-type H{sup +}-ATPase (V-ATPase), led to mis-localization of Wsc1p from the plasma membrane to the vacuole. In addition, disruption of other V-ATPase subunits (vma mutants) also caused defects of Wsc1p trafficking and vacuolar acidification similar to those seen in the vph1Δ mutant. Moreover, we found that deletion of the VPS26 gene, encoding a subunit of the retromer complex, also caused a defect in Wsc1p recycling and mis-localization of Wsc1p to the vacuole. These findings clarified the previously unidentified Wsc1p recycling pathway and requirement of V-ATPase-dependent luminal acidification for Wsc1p recycling.

60 APPLIED LIFE SCIENCES↗

Evolutionarily conserved IMPACT impairs various stress responses that require GCN1 for activating the eIF2 kinase GCN2

Highlights: •GCN1 is required for mammalian and yeast GCN2 function in a variety of conditions. •Mammalian IMPACT competes with GCN2 for GCN1 binding. •IMPACT and its yeast counterpart YIH1 downregulate GCN1-dependent GCN2 activation. -- Abstract: In response to a range of environmental stresses, phosphorylation of the alpha subunit of the translation initiation factor 2 (eIF2α) represses general protein synthesis coincident with increased translation of specific mRNAs, such as those encoding the transcription activators GCN4 and ATF4. The eIF2α kinase GCN2 is activated by amino acid starvation by a mechanism involving GCN2 binding to an activator protein GCN1, along with association with uncharged tRNA that accumulates during nutrient deprivation. We previously showed that mammalian IMPACT and its yeast ortholog YIH1 bind to GCN1, thereby preventing GCN1 association with GCN2 and stimulation of this eIF2α kinase during amino acid depletion. GCN2 activity is also enhanced by other stresses, including proteasome inhibition, UV irradiation and lack of glucose. Here, we provide evidence that IMPACT affects directly and specifically the activation of GCN2 under these stress conditions in mammalian cells. We show that activation of mammalian GCN2 requires its interaction with GCN1 and that IMPACT promotes the dissolution of the GCN2–GCN1 complex. To a similar extent as the overexpression of YIH1, overexpression of IMPACT in yeast cells inhibited growth under all stress conditions that require GCN2 and GCN1 for cell survival, including exposure to acetic acid, high levels of NaCl, H{sub 2}O{sub 2} or benomyl. This study extends our understanding of the roles played by GCN1 in GCN2 activation induced by a variety of stress arrangements and suggests that IMPACT and YIH1 use similar mechanisms for regulating this eIF2α kinase.

60 APPLIED LIFE SCIENCES↗

Mutational definition of binding requirements of an hnRNP-like protein in Arabidopsis using fluorescence correlation spectroscopy

Highlights: • We use FCS to investigate binding site requirements for the hnRNP-like protein AtGRP7. • We identify three nucleotides critical for AtGRP7 binding to its own intron. • Mutation of the conserved R{sup 49} abolishes binding altogether. • The paralogue AtGRP8 binds to an overlapping motif with different sequence requirement. • The glycine-rich stretch of a plant hnRNP-like protein contributes to binding. - Abstract: Arabidopsis thaliana glycine-rich RNA binding protein 7 (AtGRP7) is part of a negative feedback loop through which it regulates alternative splicing and steady-state abundance of its pre-mRNA. Here we use fluorescence correlation spectroscopy to investigate the requirements for AtGRP7 binding to its intron using fluorescently-labelled synthetic oligonucleotides. By systematically introducing point mutations we identify three nucleotides that lead to an increased K{sub d} value when mutated and thus are critical for AtGRP7 binding. Simultaneous mutation of all three residues abrogates binding. The paralogue AtGRP8 binds to an overlapping motif but with a different sequence preference, in line with overlapping but not identical functions of this protein pair. Truncation of the glycine-rich domain reduces the binding affinity of AtGRP7, showing for the first time that the glycine-rich stretch of a plant hnRNP-like protein contributes to binding. Mutation of the conserved R{sup 49} that is crucial for AtGRP7 function in pathogen defence and splicing abolishes binding.

60 APPLIED LIFE SCIENCES↗

Production of ABA responses requires both the nuclear and cytoplasmic functional involvement of PYR1

Abscisic acid (ABA) enhances stress tolerant responses in plants against unfavorable environmental conditions. In Arabidopsis, ABA promotes interactions between PYR/PYL/RCARs and PP2C, thereby allowing SnRK2s to phosphorylate downstream components required for the regulation of gene expression or for gating ion channels. Because PYR1 is known to localize to nucleus and cytoplasm it is a question whether nuclear or cytoplasmic PYR1 confer different functions to the ABA signaling pathway, as has been previously shown for regulatory proteins. In order to answer this question, transgenic lines expressing nuclear PYR1 were generated in an ABA insensitive mutant background. Enforced nuclear expression of PYR1 was examined by confocal microscopy and western blot analysis. Physiological analyses of the transgenic lines demonstrated that nuclear PYR1 is sufficient to generate ABA responses, such as, the inhibition of seed germination, root growth inhibition, the induction of gene expression, and stomatal closing movement. However, for the full recovery of ABA responses in the mutant background cytoplasmic PYR1 was required. The study suggests both nuclear and cytoplasmic PYR1 participate in the control of ABA signal transduction. - Highlights: • Nuclear and cytoplasmic functions of PYR1 were studied in the mutant which lacked majority of ABA responses. • Nuclear PYR1 reconstituted partially the ABA responses during seed germination, root growth, and guard cell movement. • Both the nuclear and cytoplasmic functions of PYR1 were required for the full generation of ABA responses.

60 APPLIED LIFE SCIENCES↗

Differential Etv2 threshold requirement for endothelial and erythropoietic development

Endothelial and erythropoietic lineages arise from a common developmental progenitor. Etv2 is a master transcriptional regulator required for the development of both lineages. However, the mechanisms through which Etv2 initiates the gene-regulatory networks (GRNs) for endothelial and erythropoietic specification and how the two GRNs diverge downstream of Etv2 remain incompletely understood. Here, by analyzing a hypomorphic Etv2 mutant, we demonstrate different threshold requirements for initiation of the downstream GRNs for endothelial and erythropoietic development. We show that Etv2 functions directly in a coherent feedforward transcriptional network for vascular endothelial development, and a low level of Etv2 expression is sufficient to induce and sustain the endothelial GRN. In contrast, Etv2 induces the erythropoietic GRN indirectly via activation of Tal1, which requires a significantly higher threshold of Etv2 to initiate and sustain erythropoietic development. These results provide important mechanistic insight into the divergence of the endothelial and erythropoietic lineages.

59 BASIC BIOLOGICAL SCIENCES↗

VRC34-Antibody Lineage Development Reveals How a Required Rare Mutation Shapes the Maturation of a Broad HIV-Neutralizing Lineage

Rare mutations have been proposed to restrict the development of broadly neutralizing antibodies against HIV-1, but this has not been explicitly demonstrated. We hypothesized that such rare mutations might be identified by comparing broadly neutralizing and non-broadly neutralizing branches of an antibody-developmental tree. Because sequences of antibodies isolated from the fusion peptide (FP)-targeting VRC34-antibody lineage suggested it might be suitable for such rare mutation analysis, we carried out next-generation sequencing (NGS) on B cell transcripts from donor N123, the source of the VRC34 lineage, and functionally and structurally characterized inferred intermediates along broadly neutralizing and poorly neutralizing developmental branches. The broadly neutralizing VRC34.01 branch required the rare heavy-chain mutation Y33P to bind FP, whereas the early bifurcated VRC34.05 branch did not require this rare mutation and evolved less breadth. Our results demonstrate how a required rare mutation can restrict development and shape the maturation of a broad HIV-1-neutralizing antibody lineage.

59 BASIC BIOLOGICAL SCIENCES↗

Energy storage requirements of dc microgrids with high penetration renewables under droop control

Energy storage is a important design component in microgrids with high penetration renewable sources to maintain the system because of the highly variable and sometimes stochastic nature of the sources. Storage devices can be distributed close to the sources and/or at the microgrid bus. In addition, storage requirements can be minimized with a centralized control architecture, but this creates a single point of failure. Distributed droop control enables a completely decentralized architecture but, the energy storage optimization becomes more difficult. Our paper presents an approach to droop control that enables the local and bus storage requirements to be determined. Given a priori knowledge of the design structure of a microgrid and the basic cycles of the renewable sources, we found that the droop settings of the sources are such that they minimize both the bus voltage variations and overall energy storage capacity required in the system. This approach can be used in the design phase of a microgrid with a decentralized control structure to determine appropriate droop settings as well as the sizing of energy storage devices.

25 ENERGY STORAGE↗

Parameter dependencies of the experimental nitrogen concentration required for detachment on ASDEX Upgrade and JET

While current tokamak experiments are beginning to use real-time feedback control systems to manage the plasma exhaust, future tokamaks still require validation of theoretical models used to predict the threshold impurity concentration required to sufficiently reduce the power and particle fluxes to the divertor. This work exploits new spectroscopic measurements of the divertor nitrogen concentration, c n , in partially detached N 2 -seeded H-mode plasmas on ASDEX Upgrade (AUG) and JET with the ITER-Like Wall (JET-ILW) to test the parameter dependencies of the power flowing to the outer divertor, P div,outer , and the separatrix electron density, n e.sep . A least-squares regression of the AUG measurements demonstrates that the threshold c n required for detachment scales as $c_n∝P^{1.19±0.32}_{div,outer}n^{-2.77±0.36}_{e,sep}$. This scaling of n e,sep is also consistent with the measurements from JET which, at constant P div,outer , show $c_n∝n^{-2.43±0.27}_{e,sep}$. The dependencies of P div,outer and n e.sep is demonstrated over at least a factor of two change in both parameters and indicates a stronger dependence on n e.sep in comparison to the Lengyel model, which could be due to the assumption in this model that the heat flux channel width is independent of density. This first assessment of detachment scaling with impurity seeding highlights the need for further analysis of the systematic uncertainties of the measurement and more consistent scenarios from more tokamaks to investigate the machine size scaling.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Plakophilin3 loss leads to an increase in lipocalin2 expression, which is required for tumour formation

Highlights: • Plakophilin3 loss leads to an increase in LCN2 expression in multiple cell types. • The increase in LCN2 expression is dependent on the activity of p38 MAPK. • Plakophilin3 inhibits LCN2 expression by restricting p38 MAPK to the cytoplasm. • The increase in LCN2 expression is required for tumour formation upon plakophilin3 loss. An increase in tumour formation and metastasis are observed upon plakophilin3 (PKP3) loss. To identify pathways downstream of PKP3 loss that are required for increased tumour formation, a gene expression analysis was performed, which demonstrated that the expression of lipocalin2 (LCN2) was elevated upon PKP3 loss and this is consistent with expression data from human tumour samples suggesting that PKP3 loss correlates with an increase in LCN2 expression. PKP3 loss leads to an increase in invasion, tumour formation and metastasis and these phenotypes were dependent on the increase in LCN2 expression. The increased LCN2 expression was due to an increase in the activation of p38 MAPK in the HCT116 derived PKP3 knockdown clones as LCN2 expression decreased upon inhibition of p38 MAPK. The phosphorylated active form of p38 MAPK is translocated to the nucleus upon PKP3 loss and is dependent on complex formation between p38 MAPK and PKP3. WT PKP3 inhibits LCN2 reporter activity in PKP3 knockdown cells but a PKP3 mutant that fails to form a complex with p38 MAPK cannot suppress LCN2 promoter activity. Further, LCN2 expression is decreased upon loss of p38β, but not p38α, in the PKP3 knockdown cells. These results suggest that PKP3 loss leads to an increase in the nuclear translocation of p38 MAPK and p38β MAPK is required for the increase in LCN2 expression.

60 APPLIED LIFE SCIENCES↗

Prototyping and validating requirements of radiation and nuclear emergency plan simulator

Organizational incapability in developing unrealistic, impractical, inadequate and ambiguous mechanisms of radiological and nuclear emergency preparedness and response plan (EPR) causing emergency plan disorder and severe disasters. These situations resulting from 65.6% of poor definition and unidentified roles and duties of the disaster coordinator. Those unexpected conditions brought huge aftermath to the first responders, operators, workers, patients and community at large. Hence, in this report, we discuss prototyping and validating of Malaysia radiation and nuclear emergency preparedness and response plan simulation model (EPRM). A prototyping technique was required to formalize the simulation model requirements. Prototyping as systems requirements validation was carried on to endorse the correctness of the model itself against the stakeholder’s intensions in resolving those organizational incapability. We have made assumptions for the proposed emergency preparedness and response model (EPRM) through the simulation software. Those assumptions provided a twofold of expected mechanisms, planning and handling of the respective emergency plan as well as in bringing off the hazard involved. This model called RANEPF (Radiation and Nuclear Emergency Planning Framework) simulator demonstrated the training emergency response perquisites rather than the intervention principles alone. The demonstrations involved the determination of the casualties’ absorbed dose range screening and the coordination of the capacity planning of the expected trauma triage. Through user-centred design and sociotechnical approach, RANEPF simulator was strategized and simplified, though certainly it is equally complex.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

Differential pumping requirements for the light-ion helicon source and heating systems of Proto-MPEX

The physics of electron and ion heating of high-density deuterium helicon plasmas (>3 × 10 19 m –3 ) in the Proto-Material Plasma Exposure Experiment linear device are under investigation. Theoretical estimates indicate that for efficient heating, discharges with very low neutral gas content ( &laquo; 0.1 Pa) in the heating sections are required to minimize collisional losses and charge exchange interactions with neutrals. However, this requirement is typically not compatible with the neutral gas pressures (1–2 Pa) commonly used in high-density, light-ion helicon sources. To satisfy these competing requirements, differential pumping techniques are needed. Here, results are presented that demonstrate the production of high-density discharges (2–6 × 10 19 m –3 ) with very low neutral gas content (<0.01 Pa) and high degree of ionization (>75%) in the heating sections. Results indicate that the best fueling location is upstream of the plasma source. We elaborate on the key aspects that must be considered to produce these discharges: (1) fueling location, radio-frequency pulse length, and magnetic field configuration, (2) flow rate and timing of the gas injection, and (3) use of conductance-limiting elements.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗