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At least 73 records · Page 4

The Omics of Stem Cell Mediated Regeneration: A Pilot Single Cell RNA-Seq Study of Mechanotransduction

Mechanical forces are potent modulators of stem cell based tissue regenerative mechanisms, inducing cell fate decisions and tissue specific commitment. A unique platform for investigating mechanotransduction is spaceflight, where microgravity and altered fluid mechanics provide a loading-null experimental condition. Seminal investigations of regenerative capacity in a wholly regenerative species, the newt model, and in a variety of totipotent and adult stem cell populations have demonstrated the detrimental effects of unloading on maintenance of stem cell based regeneration. Of particular interest is the observation that unloading interferes with the transition of stem cell pools from proliferative state to differentiation commitment. In this work we sought to test the hypothesis that gravity mechanotransduction regulates stem cell tissue regenerative processes by modulating stem cell proliferation and differentiation fates at specific cell cycle stages. To do this, clonally-derived ESCs were plated on a collagen matrix and expanded for 36 hours before re-plating on a non-adherent culture dish in the absence of leukemia inhibitory factor (LIF) to form spheroid aggregate EBs. After formation, the EBs were transferred to a collagen matrix coated culture dishes and given 4 days to allow implantation and outgrowth. In parallel, totipotent ESCs were plated 24 hours before mechanical stimulation on collagen matrix culture dishes in the presence of LIF to maintain totipotency and serve as un-differentiation committed controls. The EBs and ESCs were then subjected to either a 60 minute pulse of gravity (static loading) or 60 minutes of cyclic stretch (dynamic loading) mechanotransduction. Six hours post-stimulation, we used a 10X Genomics Single Cell controller to generate bar-coded single cell Illumina libraries and sequenced expressomes for 5,000 static loaded cells, representative of a change in gravity mechanotransduction, 5,000 dynamic loaded cells, representative of tissue loading associate with physiologic function, and 5,000 unstimulated 1g control cells. The comparison of these 3 libraries by cluster assignment based on like gene expression patterns show substantial alteration in cluster geometry due to mechanical loading. Specifically the mechanically loaded EB outgrowth cells to retain potency markers (PAX6, SOX2, CD34) and suppress early commitment markers (Dhh, VCAN, Igf1). Whereas the EBs cultured under the non-stimulated conditions display clear departure from the ESC expressome with lineage commitment markers upregulated and several tissue specific markers being expressed (BMP "early musculoskeletal development, Mesp1" early cardiovascular cell lineage). These markers are not seen in the mechano-stimulated cultures or the totipotent ESC cultures. Comparison of like clusters between our experimental conditions revealed an array of regenerative and stem cell genes are significantly mechano-regulated. Of particular importance CDKN1a/p21, a gene shown by previous investigation of our research team to be significantly upregulated in unloading, was suppressed in the static and dynamic loaded EBS. In addition to CDKN1a/p21 many genes related to cell cycle and transitory differentiation markers had elevated expression in the mechano-stimulated EBs, but surprisingly these trends were not observed in the ESC cultures. This study is the first of its kind investigating for mechano-signaling and mechano-regulated pathways, and has alre

Single cell RNA-sequecing↗

CDKN1A/P21’S Role in Osteoprogenitor Fate Regulation in Response to Mechanical Loading: A Single Cell Resolution Investigation

Forces generated by gravity in load-bearing tissues such as bone marrow promote stem cell-based tissue-regenerative processes by increasing proliferation and differentiation of tissue progenitors. Conversely, in microgravity, mouse bone marrow mesenchymal and hematopoietic precursors down-regulate differentiation markers and up-regulate stemness maintenance genes. During microgravity, these transcriptomic changes are associated with Cdkn1a overexpression in osteoprogenitors, and further the homozygous deletion of the CDKN1a gene in the mouse model results in elevated stem-cell based regeneration of severed digits and ear-hole punches. In this work we sought to test the hypothesis that gravity mechanotransduction regulates mesenchymal stem cell derived osteoprogenitor based osteogenesis by modulating proliferation and differentiation fates at specific cell cycle stages via a p21/CDKN1a-regulated mechanism.. To test our hypothesis, we isolated primary mouse bone marrow osteoprogenitors from Cdkn1a-/- (null) or wildtype mice, cultured the cells for 7-days unloaded in osteogenic differentiation media, to allow commitment of the adherent cell population, and subsequently, cultured an additional 48-hours under cyclic stretch or static control loading conditions. After 48-hours of dynamic mechanical loading, we used a 10X Genomics Chromium/Single Cell controller to generate bar-coded single cell Illumina libraries and sequenced expressomes for 6,000 static control and 6,000 cyclic stretch cells for each genotype. Stage-specific single cell analyses show Cdkn1a-/- osteogenic cultures are more differentiated with fewer progenitors, and that cyclic stretch further promotes the conversion of progenitors to osteoblasts in both wildtype and Cdkn1a-/- backgrounds. The proportion of late mineralizing osteoblasts doubled in Cdkn1a-/- cultures, but is not affected by load. Finally, single cell expression of Cdkn1a, in the wildtype cells, is most strongly suppressed by cyclic stretch both in early and late osteoblasts, and minimally in the progenitor population. Collectively, our results support the hypothesis that Cdkn1a constitutively plays a mechano-reversible anti-proliferative role during stem cell-based bone tissue regeneration, and suggests a new molecular target to counter regenerative deficits caused by disuse.

Eduardo Almeida↗

Managing work flow and logistic at waste processing facilities using the waste compliance and tracking system - 15392

Work flow and logistics at a busy radioactive waste processing complex or permitted treatment, storage, and disposal facility (TSDF) can pose a monumental challenge. An efficient operation requires good communication between engineering, logistics, and waste operators. The Los Alamos National Laboratory (LANL) has implemented an application that facilitates seamless communication of work flow requirements, between engineering, logistics, and waste operations personnel in the field. The Waste Compliance and Tracking System (WCATS), a comprehensive waste management system designed at LANL, provides four key elements to support logistics: (a) the container work path, (b) directed unit operation requests, (c) notification of pending operations, and (d) real-time status tools for office and field workers. The combination of these elements allows everyone to function as a team, with a shared knowledge of what is needed to accomplish facility objectives. The first element is like a road map indicating which unit operations are needed and in what sequence they should be performed; this is referred to as a 'work path'. It automatically routes a waste item or container from operation to operation. For example, a given work path might indicate that waste items require radiological survey, radioassay, and a technical review, in that order. When the work path is assigned to a container, the system enforces the correct order of operations. Directed unit operation 'requests', the second element, fills a gap between pre-planned, formal 'work path' activities and the need for flexibility to direct preparatory operations or deal with unexpected issues, like staging or mining containers in preparation for a shipment or transferring containers from one storage unit to another to clear a path for building maintenance. Logistics engineers prepare these 'requests', and the system monitors the status and completion of the work. The third element, notifications, allows team members to subscribe to and receive notice that an operation is pending, initiated, or complete. This functionality supports a range of communication options, including e-mail, cell phone, pager, or application in-box messaging. Notifications allow waste operations personnel to multi-task, and remain in the loop on the status of facility-wide activities. Finally, the fourth element provides tools that communicate work path pending operations and directed requests to field workers on their mobile computer, allowing them to mine or pick containers from storage units, and leverage the capabilities of a bar-code scanner to avoid selection mistakes. Oversight and summary tools are provided, allowing logistics personnel to monitor work path and request task status, identify work-flow bottlenecks, and maximize the efficiency of waste operations. The Waste Compliance and Tracking System (WCATS) provides an innovative, comprehensive set of tools that allow large-scale radioactive waste processing facilities to manage logistics, maximize efficient use of the workforce, and create a team environment, where everyone from management to the waste handlers have an understanding of the work load and what needs to be accomplished. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

A Study on the Comparison of International WTS for Integrated Management of Radioactive Waste - 19034

Domestic radioactive waste is generated by nuclear power plants, the Korea Atomic Energy Research Institute and KEPCO Nuclear Fuel Company. These nuclear energy agencies have their own waste tracking system. To track waste more efficiently, it is necessary to build an integrated tracking system from the viewpoint of the final disposer. As for the korea Atomic Energy Research Institute, the decommissioning waste due to the decommissioning of research reactors No. 1 and 2, and the low and intermediate-level radioactive waste, generated by the operation of the High Flux Advanced Neutron Application Reactor(HANARO), account for the most. The solid waste from each facility and laboratory is sorted and collected according to radiation levels and the contents. For starters, according to the contents, the waste is divided into combustible, noncombustible compressible and non-compressible waste. The combustible waste, generated by the Korea Atomic Energy Research Institute, is treated according to the low and intermediate-level radioactive waste delivery regulation of the Nuclear Safety and Security Commission and the acceptance criteria of the disposal facility. KEPCO Nuclear Fuel Company is making the nuclear fuel used in nuclear power plants. Radioactive waste is generated in the nuclear fuel manufacturing process, and it is classified into 8 types of waste according to the solid waste management procedure. Most of the radioactive waste, generated while KEPCO Nuclear Fuel Company processes the light-water reactor and heavy-water reactor nuclear fuel, is classified as low and intermediate-level radioactive waste. The radioactive liquid waste, generated in the radiation controlled area in nuclear fuel processing facilities, is divided into low-concentration liquid waste and high-concentration liquid waste. The miscellaneous solids, generated in the Hanbit NPP of Korea Hydro and Nuclear Power Co., Ltd., are classified in consideration of the physical properties, density and amount of the waste, and substances subject to restricted disposal. This paper investigated information on the structure, functions and maintenance of the systems implemented and operated by individual countries and IAEA, and the types of managed data, and analyzed the detailed configuration of the system and the characteristics of DBMS (Data Base Management System), and discovered during the analysis process the information on the waste management in each country or institution. This paper investigated the classification standards and treatment methods according to the characteristics of the radioactive substances handled by each agency. These classification and treatment methods can be applied to the database when the radioactive waste tracking system is implemented in the future. Also, this paper analyzed the radioactive waste tracking system of each country. Most overseas waste tracking systems are configured based on the ORACLE DBMS. The ORACLE DBMS is highly secure, and can manage large-scale databases. Also, Germany's ReVK and US IWTS are using the Bar-Code system that can track and manage all waste data, generated during decommissioning or transportation, in real time, and the real-time tracking system of web-based US CID can simply create and output reports quickly. If the advantages of overseas waste tracking systems are added to the domestic integrated waste tracking system, more efficient and safer management of radioactive waste will be possible.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Data compression system

A data compression system is described in which TV PCM data for each line scan is received in the form of a succession of multibit pixel words. All or selected bits of each word are compressed by providing difference values between successive pixel words and coding the difference values of a selected number of pixel words forming a block into a fundamental sequence (FS). The FS, based on its length and the number of words per block, is either transmitted as the compressed data or is used to generate a code FS or its complement is used to generate a code FS bar. When the code FS is generated, its length is compared with the original block PCM and only if the former is the shorter of the two is the code transmitted. Selected bits per pixel word may be compressed, while the remaining bits may be transmitted directly, or some of them may be omitted altogether.

Rice, R. F.↗

Mapping DNA polymerase errors by single-molecule sequencing

Genomic integrity is compromised by DNA polymerase replication errors, which occur in a sequence-dependent manner across the genome. Accurate and complete quantification of a DNA polymerase's error spectrum is challenging because errors are rare and difficult to detect. We report a high-throughput sequencing assay to map in vitro DNA replication errors at the single-molecule level. Unlike previous methods, our assay is able to rapidly detect a large number of polymerase errors at base resolution over any template substrate without quantification bias. To overcome the high error rate of high-throughput sequencing, our assay uses a barcoding strategy in which each replication product is tagged with a unique nucleotide sequence before amplification. Here, this allows multiple sequencing reads of the same product to be compared so that sequencing errors can be found and removed. We demonstrate the ability of our assay to characterize the average error rate, error hotspots and lesion bypass fidelity of several DNA polymerases.

60 APPLIED LIFE SCIENCES↗

Frequency monitoring a LIDAR transmitter for pulse-by-pulse analysis and control

An electronic technique is described whereby the heterodyne mixing frequency of a pulsed LIDAR transmitter is displayed on a color-coded linear bar graph. The technique is of great utility for manually fine tuning the laser output frequency when the use of conventional frequency counters and spectrum analyzers is impractical in a rapidly firing system.

Esproles, Carlos↗

Using Modified Fagan Inspections to Control Rapid System Development

The Jet Propulsion Laboratory (JPL) has been developing new approaches to software and system development to shorten life cycle time and reduce total life-cycle cost, while maintaining product quality. One such approach has been taken by the Just-In-Time (JIT) Materiel Acquisition System Development Project.

JIT Implementing Radio Frequency (RF) Bar Code dev↗

Microbial certification of the MER spacecraft

We conclude in this paper that a combination of Dry Heat Microbrial Reduction and control measures during complex mechanical assembly processes can result in a total spore bioburden that meets requirements.

spore bioburden MER spacecraft bacteria bar code↗

Verification of Advective Bar Elements Implemented in the Aria Thermal Response Code.

A verification effort was undertaken to evaluate the implementation of the new advective bar capability in the Aria thermal response code. Several approaches to the verification process were taken : a mesh refinement study to demonstrate solution convergence in the fluid and the solid, visually examining the mapping of the advective bar element nodes to the surrounding surfaces, and a comparison of solutions produced using the advective bars for simple geometries with solutions from commercial CFD software . The mesh refinement study has shown solution convergence for simple pipe flow in both temperature and velocity . Guidelines were provided to achieve appropriate meshes between the advective bar elements and the surrounding volume. Simulations of pipe flow using advective bars elements in Aria have been compared to simulations using the commercial CFD software ANSYS Fluent (r) and provided comparable solutions in temperature and velocity supporting proper implementation of the new capability. Verification of Advective Bar Elements iv Acknowledgements A special thanks goes to Dean Dobranich for his guidance and expertise through all stages of this effort . His advice and feedback was instrumental to its completion. Thanks also goes to Sam Subia and Tolu Okusanya for helping to plan many of the verification activities performed in this document. Thank you to Sam, Justin Lamb and Victor Brunini for their assistance in resolving issues encountered with running the advective bar element model. Finally, thanks goes to Dean, Sam, and Adam Hetzler for reviewing the document and providing very valuable comments.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Validation and benchmarking of two particle-in-cell codes for a glow discharge

The two particle-in-cell codes EDIPIC and LSP are benchmarked and validated for a parallel-plate glow discharge in helium, in which the axial electric field had been carefully measured, primarily to investigate and improve the fidelity of their collision models. The scattering anisotropy of electron-impact ionization, as well as the value of the secondary-electron emission yield, are not well known in this case. The experimental uncertainty for the emission yield corresponds to a factor of two variation in the cathode current. If the emission yield is tuned to make the cathode current computed by each code match the experiment, the computed electric fields are in excellent agreement with each other, and within about 10% of the experimental value. The non-monotonic variation of the width of the cathode fa ll with the applied voltage seen in the experiment is reproduced by both codes. The electron temperature in the negative glow is within experimental error bars for both codes, but the density of slow trapped electrons is underestimated. A more detailed code comparison don e for several synthetic cases of electron-beam injection into helium gas shows that the codes are in excellent agreement for ionization rate, as well as for elastic and excitation collisions with isotropic scattering pattern. The remaining significant discrepancies between the two codes are due to differences in their electron binary-collision models, and for anisotropic scattering due to elastic and excitation collisions.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Benchmark Calculation for the Watts Bar Unit 1 Cycles 1-3 Using the SCALE 6.3/Polaris–PARCS v3.4.2 Code Package

Benchmark calculations for the Watts Bar Unit 1 cycles 1–3 were performed to validate the SCALE 6.3/Polaris–PARCS v3.4.2 with the ENDF/B-VII.1 56-group library by comparing the simulated results with the measured data. The benchmark results are used in evaluating uncertainties of the SCALE/Polaris–PARCS code package for pressurized water reactor physics analysis for key nuclear parameters such as reactivity, control bank worth, temperature coefficients, and pin and assembly power peaking factors. This report details plant and fuel design specifications and input data for SCALE/Polaris, GenPMAXS, and PARCS. Additional details are provided for the input and output files produced for the benchmark calculations. The benchmark results were summarized such that they can be used in evaluating uncertainties for key nuclear parameters with other benchmark results.

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗

Verification of Advective Bar Elements Implemented in the Sierra/Aria Thermal Response Code

A thorough code verification effort has been performed on a reduced order, finite element model for 1D fluid flow convectively coupled with a 3D solid, referred to as the 'advective bar' model. The purpose of this effort was to provide confidence in the proper implementation of this model within the SIERRA/Aria thermal response code at Sandia National Laboratories. The method of manufactured solutions is applied so that the order of convergence in error norms for successively refined meshes and timesteps is investigated. Potential pitfalls that can lead to a premature evaluation of the model's implementation are described for this verification approach when applied to this unique model. As a result, through observation of the expected order of convergence, these verification tests provide evidence of proper implementation of the model within the codebase.

42 ENGINEERING↗

All Hands On Deck: Raising the Bar on Whole Building Performance-Based Code Compliance and Above-Code Programs

In 2019 a large-scale national effort was launched with support from U.S. Department of Energy, Pacific Northwest National Laboratory and Northwest Energy Efficiency Alliance (NEEA). The project aimed to develop a long-term vision and a roadmap for achieving a practical application of whole building performance-based code and above code program compliance in commercial buildings. The work was aided by engaging over 70 stakeholders across the country representing jurisdictions, above-code programs such as LEED and ENERGY STAR Multifamily Program, members of the ASHRAE Standard 90.1 and other Standard committees, IBPSA and software developers. The performance path requires whole building energy modeling and is gaining momentum. Many see it as the future of the commercial energy codes and the main pathway for achieving zero energy buildings. The paper describes the findings from a stakeholder survey funded by NEEA on the current state of performance-based path, its market penetration, trends, implementation practices and adoption challenges. It discusses the patterns revealed by experience of the states with decades-long history of performance-based compliance such as Florida and California and opportunities for taking advantage of the synergies between code and above-code programs. In addition, the paper provides an overview of the tools and resources developed as part of the effort including the compliance form for ASHRAE Standard 90.1 Section 11 and Appendix G, modeling submittal review manual, and modeler and reviewer qualification requirements. The tools and resources would help remove market barriers, deliver immediate benefits to jurisdictions and rating authorities, and achieve energy efficiency through whole-building performance path.

TSPR, Performance Based Complinace, Washington Sta↗

Design and Control of Compliant Tensegrity Robots Through Simulation and Hardware Validation

To better understand the role of tensegrity structures in biological systems and their application to robotics, the Dynamic Tensegrity Robotics Lab at NASA Ames Research Center has developed and validated two different software environments for the analysis, simulation, and design of tensegrity robots. These tools, along with new control methodologies and the modular hardware components developed to validate them, are presented as a system for the design of actuated tensegrity structures. As evidenced from their appearance in many biological systems, tensegrity ("tensile-integrity") structures have unique physical properties which make them ideal for interaction with uncertain environments. Yet these characteristics, such as variable structural compliance, and global multi-path load distribution through the tension network, make design and control of bio-inspired tensegrity robots extremely challenging. This work presents the progress in using these two tools in tackling the design and control challenges. The results of this analysis includes multiple novel control approaches for mobility and terrain interaction of spherical tensegrity structures. The current hardware prototype of a six-bar tensegrity, code-named ReCTeR, is presented in the context of this validation.

Robotics↗

Protostellar hydrodynamics: Constructing and testing a spacially and temporally second-order accurate method. 2: Cartesian coordinates

In Boss & Myhill (1992) we described the derivation and testing of a spherical coordinate-based scheme for solving the hydrodynamic equations governing the gravitational collapse of nonisothermal, nonmagnetic, inviscid, radiative, three-dimensional protostellar clouds. Here we discuss a Cartesian coordinate-based scheme based on the same set of hydrodynamic equations. As with the spherical coorrdinate-based code, the Cartesian coordinate-based scheme employs explicit Eulerian methods which are both spatially and temporally second-order accurate. We begin by describing the hydrodynamic equations in Cartesian coordinates and the numerical methods used in this particular code. Following Finn & Hawley (1989), we pay special attention to the proper implementations of high-order accuracy, finite difference methods. We evaluate the ability of the Cartesian scheme to handle shock propagation problems, and through convergence testing, we show that the code is indeed second-order accurate. To compare the Cartesian scheme discussed here with the spherical coordinate-based scheme discussed in Boss & Myhill (1992), the two codes are used to calculate the standard isothermal collapse test case described by Bodenheimer & Boss (1981). We find that with the improved codes, the intermediate bar-configuration found previously disappears, and the cloud fragments directly into a binary protostellar system. Finally, we present the results from both codes of a new test for nonisothermal protostellar collapse.

Myhill, Elizabeth A.↗