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At least 1,297 records · Page 72

Mutations That Alter the Bacterial Cell Envelope Increase Lipid Production

ABSTRACT Lipids from microbes offer a promising source of renewable alternatives to petroleum-derived compounds. In particular, oleaginous microbes are of interest because they accumulate a large fraction of their biomass as lipids. In this study, we analyzed genetic changes that alter lipid accumulation in Rhodobacter sphaeroides . By screening an R. sphaeroides Tn 5 mutant library for insertions that increased fatty acid content, we identified 10 high-lipid (HL) mutants for further characterization. These HL mutants exhibited increased sensitivity to drugs that target the bacterial cell envelope and changes in shape, and some had the ability to secrete lipids, with two HL mutants accumulating ~60% of their total lipids extracellularly. When one of the highest-lipid-secreting strains was grown in a fed-batch bioreactor, its lipid content was comparable to that of oleaginous microbes, with the majority of the lipids secreted into the medium. Based on the properties of these HL mutants, we conclude that alterations of the cell envelope are a previously unreported approach to increase microbial lipid production. We also propose that this approach may be combined with knowledge about biosynthetic pathways, in this or other microbes, to increase production of lipids and other chemicals. IMPORTANCE This paper reports on experiments to understand how to increase microbial lipid production. Microbial lipids are often cited as one renewable replacement for petroleum-based fuels and chemicals, but strategies to increase the yield of these compounds are needed to achieve this goal. While lipid biosynthesis is often well understood, increasing yields of these compounds to industrially relevant levels is a challenge, especially since genetic, synthetic biology, or engineering approaches are not feasible in many microbes. We show that altering the bacterial cell envelope can be used to increase microbial lipid production. We also find that the utility of some of these alterations can be enhanced by growing cells in bioreactor configurations that can be used industrially. We propose that our findings can inform current and future efforts to increase production of microbial lipids, other fuels, or chemicals that are currently derived from petroleum.

59 BASIC BIOLOGICAL SCIENCES↗

CRISPR-prime editing, a versatile genetic tool to create specific mutations with a single nucleotide resolution in Leptospira

ABSTRACT Leptospirosis, caused by pathogenic bacteria from the genusLeptospira, is a global zoonosis responsible for more than one million human cases and 60,000 deaths annually. The disease also affects many domestic animal species. Historically, genetic manipulation ofLeptospirahas been difficult to perform, resulting in limited knowledge on pathogenic mechanisms of disease and the identification of virulence factors. The application of CRISPR/Cas9 and its variations have helped fill these gaps but the generation of knockout mutants remains challenging because double-strand breaks (DSBs) inflicted by Cas9 nuclease are lethal toLeptospiracells. The novel CRISPR prime editing (PE) strategy is the first precise genome-editing technology that allows deletions, insertions, and base substitutions without introducing DSBs. This revolutionary technique utilizes a nickase Cas9 that cleaves a single strand of DNA, coupled with an engineered reverse transcriptase and a modified single-guide RNA (termed prime editing guide RNA) containing an extended 3′ end with the desired edits. We demonstrate the application of CRISPR-PE in both saprophytic and pathogenicLeptospirafrom multiple species and serovars by introducing deletions or insertions into target DNA with a remarkable precision of just one nucleotide. Additionally, we demonstrate the ability to genetically manipulateLeptospira borgpetersenii, a prevalent pathogenic species of humans, domestic cattle, and wildlife animals. Rapid plasmid loss by mutated strains in liquid culture allows for the generation of knockout strains without selective markers, which can be readily used to elucidate virulence factors and develop optimized bacterin and/or live vaccines against leptospirosis. IMPORTANCE Leptospirosis is a geographically widespread bacterial zoonosis. Genetic manipulation of pathogenicLeptospiraspp. has been laborious and difficult to perform, limiting our ability to understand how leptospires cause disease. The application of the CRISPR/Cas9 system toLeptospiraenhanced our ability to generate knockdown and knockout mutants; however, the latter remains challenging. Here, we demonstrate the application of the CRISPR prime editing technique inLeptospira, allowing the generation of knockout mutants in several pathogenic species, with mutations comprising just a single nucleotide resolution. Notably, we generated a mutant in theLeptospira borgpeterseniibackground, a prevalent pathogenic species of humans and cattle. Our application of this method opens new avenues for studying pathogenic mechanisms ofLeptospiraand the identification of virulence factors across multiple species. These methods can also be used to facilitate the generation of marker-less knockout strains for updated and improved bacterin and/or live vaccines.

Microbiology↗

Geometric pre-patterning based tuning of the period doubling onset strain during thin film wrinkling

Wrinkling of supported thin films is an easy-to-implement and low-cost fabrication technique for generation of stretch-tunable periodic micro and nano-scale structures. However, the tunability of such structures is often limited by the emergence of an undesirable period doubled mode at high strains. Predictively tuning the onset strain for period doubling via existing techniques requires one to have extensive knowledge about the nonlinear pattern formation behavior. Herein, a geometric pre-patterning based technique is introduced to delay the onset of period doubling that can be implemented to predictively tune the onset strain even with limited system knowledge. The technique comprises pre-patterning the film/base bilayer with a sinusoidal pattern that has the same period as the natural wrinkle period of the system. The effectiveness of this technique has been verified via physical and computational experiments on the polydimethylsiloxane/glass bilayer system. It is observed that the period doubling onset strain can be increased from the typical value of 20% for flat films to greater than 30% with a modest pre-pattern aspect ratio (2∙amplitude/period) of 0.15. In addition, finite element simulations reveal that (i) the onset strain can be increased up to a limit by increasing the amplitude of the pre-patterns and (ii) the delaying effect can be captured entirely by the pre-pattern geometry. As a result, one can implement this technique even with limited system knowledge, such as material properties or film thickness, by simply replicating pre-existing wrinkled patterns to generate prepatterned bilayers. Thus, geometric pre-patterning is a practical scheme to suppress period doubling that can increase the operating range of stretch-tunable wrinkle-based devices by at least 50%.

36 MATERIALS SCIENCE↗

Consortium for Verification Technology Fellowship Report.

As one recipient of the Consortium for Verification Technology (CVT) Fellowship, I spent eight days as a visiting scientist at the University of Michigan, Department of Nuclear Engineering and Radiological Sciences (NERS). During this time, I participated in multiple department and research group meetings and presentations, met with individual faculty and students, toured multiple laboratories, and taught one-half of a one-unit class on Risk Analysis in Nuclear Arms control (six 1.5 hour lectures). The following report describes some of the interactions that I had during my time as well as a brief discussion of the impact of this fellowship on members of the consortium and on me/my laboratory’s technical knowledge and network.

98 NUCLEAR DISARMAMENT, SAFEGUARDS, AND PHYSICAL P↗

Argonne Leadership Computing Facility (2018 Annual Report)

The Argonne Leadership Computing Facility enables breakthroughs in science and engineering by providing supercomputing resources and expertise to the research community. In 2018, we continued to build up programs and efforts that reinforce our overall simulation, learning, and data analysis capabilities as a facility. We also enhanced our services to better meet our users’ needs and workflow preferences.

96 KNOWLEDGE MANAGEMENT AND PRESERVATION↗

Metamaterials as a Platform for the Development of Novel Materials for Energy Applications

To explore the fundamental properties of metamaterials (MMs) / metasurfaces and their potential for control of energy at the sub‐wavelength scale in support of the mission of the Department of Energy and the office of Basic Energy Sciences. Electromagnetic metamaterials provide a platform for the discovery and design of new materials with novel structures, functions, and properties. The PI proposes to advance the knowledge base of these materials through fundamental investigations of the experimental and theoretical properties of metamaterials for the discovery, prediction and design of new materials with novel structures, functions, and properties. The proposed research activities emphasize a complete basic research program including the conceptual / computational design, fabrication / synthesis of the materials, and the characterization and analysis of their electromagnetic properties. The proposed project explores the fundamental properties of metamaterials / metasurfaces and their potential for energy applications. There are three main topics which will be investigated: 1) Dispersion engineering with metamaterials and metasurfaces, 2) Epsilon near zero metamaterial absorbers and emitters, and 3) All dielectric metamaterials. The program implements a complete basic research program consisting of theory / design, modeling, characterization, and analysis, in order to fully characterize metamaterials and metasurfaces, while at the same time minimizing iterations necessary to achieve the proposal goals.

36 MATERIALS SCIENCE↗

Factors that influence the activity of biomass-degrading enzymes in the presence of ionic liquids—a review

Ionic liquids (ILs) are seen as a more sustainable alternative to volatile organic solvents that are accelerating innovations in many industries such as energy storage, separations, and bioprocessing. The ability to effectively deconstruct lignocellulosic biomass is a significant hurdle in the biorefining/bioprocessing industry and presents limitations towards the commercial production of bioproducts (such as biofuels, biomaterials, etc. ). Certain ILs have been shown to promote effective lignin removal, cellulose recovery, and sugar yields from various biomass feedstocks such as corn stover, wheat straw, sugarcane bagasse, sorghum, switchgrass, miscanthus, poplar, pine, eucalyptus, and certain mixtures of municipal solid waste. However, these improvements are often counteracted by the limited biocompatibility of ILs, which results in an IL-induced reduction in enzyme activity and stability—an important downstream step in the conversion of biomass to biofuels/bioproducts. As a result, significant efforts have been made to discover and engineer compatible enzyme-IL systems and to improve our understanding on the effect that these ILs have on these systems. This review seeks to examine the impact of ionic liquids on enzymes involved in lignocellulosic biomass deconstruction, with a specific focus on their relevance in the context of pretreatment. Beyond presenting an overview of the ionic liquid pretreatment landscape, we outline the main factors that influence enzyme activity and stability in the presence of ILs This data is consolidated and analyzed to apply this body of knowledge towards new innovations that could lead to improvements in the processing of biomass to biofuels and bioproducts.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

The Geoengineering Model Intercomparison Project- Introduction to the Second Special Issue

The Geoengineering Model Intercomparison Project (GeoMIP) has just concluded its second special issue, which this article introduces. Here we discuss the history of GeoMIP, what was learned in the first special issue, why a second special issue was needed, the additional knowledge that was provided in that issue, and some next steps for GeoMIP and the field of geoengineering research. We focus on response and uncertainty across climate models, questions for which GeoMIP is particularly well suited.

42 ENGINEERING↗

Generic Argillite/Shale Disposal Reference Case

Radioactive waste disposal in a deep subsurface repository hosted in clay/shale/argillite is a subject of widespread interest given the desirable isolation properties, geochemically reduced conditions, and widespread geologic occurrence of this rock type (Hansen 2010; Bianchi et al. 2013). Bianchi et al. (2013) provides a description of diffusion in a clay-hosted repository based on single-phase flow and full saturation using parametric data from documented studies in Europe (e.g., ANDRA 2005). The predominance of diffusive transport and sorption phenomena in this clay media are key attributes to impede radionuclide mobility making clay rock formations target sites for disposal of high-level radioactive waste. The reports by Hansen et al. (2010) and those from numerous studies in clay-hosted underground research laboratories (URLs) in Belgium, France and Switzerland outline the extensive scientific knowledge obtained to assess long-term clay/shale/argillite repository isolation performance of nuclear waste. In the past several years under the UFDC, various kinds of models have been developed for argillite repository to demonstrate the model capability, understand the spatial and temporal alteration of the repository, and evaluate different scenarios. These models include the coupled Thermal-Hydrological-Mechanical (THM) and Thermal-Hydrological-Mechanical-Chemical (THMC) models (e.g. Liu et al. 2013; Rutqvist et al. 2014a, Zheng et al. 2014a) that focus on THMC processes in the Engineered Barrier System (EBS) bentonite and argillite host hock, the large scale hydrogeologic model (Bianchi et al. 2014) that investigates the hydraulic connection between an emplacement drift and surrounding hydrogeological units, and Disposal Systems Evaluation Framework (DSEF) models (Greenberg et al. 2013) that evaluate thermal evolution in the host rock approximated as a thermal conduction process to facilitate the analysis of design options. However, the assumptions and the properties (parameters) used in these models are different, which not only make inter-model comparisons difficult, but also compromise the applicability of the lessons learned from one model to another model. The establishment of a reference case would therefore be helpful to set up a baseline for model development. A generic salt repository reference case was developed in Freeze et al. (2013) and the generic argillite repository reference case is presented in this report. The definition of a reference case requires the characterization of the waste inventory, waste form, waste package, repository layout, EBS backfill, host rock, and biosphere. This report mainly documents the processes in EBS bentonite and host rock that are potentially important for performance assessment and properties that are needed to describe these processes, with brief description other components such as waste inventory, waste form, waste package, repository layout, aquifer, and biosphere. A thorough description of the generic argillite repository reference case will be given in Jové Colon et al. (2014).

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Quantification of Cation Sorption to Engineered Barrier Materials Under Extreme Conditions

The objective of this research is to examine mechanisms and thermodynamics of actinide sorption to engineered barrier materials (iron (oxyhydr)oxides and bentonite clay) for nuclear waste repositories under high temperature and high ionic strength conditions using a suite of macroscopic and microscopic techniques which will be coupled with interfacial reaction models. Gaining a mechanistic understanding of interfacial processes governing the sorption/sequestration of actinides at mineral-water interfaces is fundamental for the accurate prediction of actinide behavior in waste repositories. Although macroscale sorption data and various spectroscopic techniques have provided valuable information regarding speciation of actinides at solid-water interfaces, significant knowledge gaps still exist with respect to sorption mechanisms and the ability to quantify sorption, particularly at high temperatures and ionic strengths. This objective is addressed through three major tasks: (1) influence of oxidation state on actinide sorption to iron oxides and clay minerals at elevated temperatures and ionic strengths; (2) calorimetric titrations of actinide-mineral suspensions; (3) evaluation of bentonite performance under repository conditions. The results of the work will include a qualitative conceptual model and a quantitative thermodynamic speciation model describing actinide partitioning to minerals and sediments, which is based upon a mechanistic understanding of specific sorption processes as determined from both micro-scale and macroscale experimental techniques. The speciation model will be a thermodynamic aqueous and surface complexation model of actinide interactions with mineral surfaces that is self-consistent with macroscopic batch sorption data, calorimetric and potentiometric titrations, X-ray absorption Spectroscopy (XAS, mainly Extended X-ray Absorption Fine Structure (EXAFS)), and electron microscopy analyses. The novelty of the proposed work lies largely in the unique system conditions which will be examined (i.e. elevated temperature and ionic strength) and the manner in which the surface complexation model will be developed in terms of specific surface species identified using XAS. These experiments will thus provide a fundamental understanding of the chemical and physical processes occurring at the solid-solution interface under expected repository conditions. Additionally, the focus on thermodynamic treatment of actinide ion interactions with minerals as proposed will provide information on the driving forces involved and contribute to the overall understanding of the high affinity many actinide ions have for oxide surfaces. The utility of this model will be demonstrated in this work through a series of advective and diffusive flow experiments.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Addressing Human and Organizational Factors In Nuclear Industry Modernization: A Sociotechnically-Based Strategic Framework

The modernization of nuclear power plants will require an advanced concept of operations, involving an integrated set of tightly coupled systems in which all stakeholders act in a coordinated manner. For this modernization effort to be enabled, we developed a human and organizational factors approach, based on a broad sociotechnical framework. Starting from core human factors principles, we conducted a literature review of the methods and approaches relevant to the modernization problem. These included core disciplines such as cognitive systems engineering, systems theoretic accident modeling and processes (STAMP), human systems integration, resilience engineering, and macroergonomics but also related topics of safety culture and organizational change. From this literature, we developed a conceptual framework centered around the work system with its four interacting components: people, technology, process, and governance. In an effective work system, these four components are jointly optimized according to three systems criteria: efficiency, effectiveness, and safety. System failure may result from excessive emphasis on any one criterion. The actual work of attaining joint optimization in a given work system can be accomplished by utilizing three high level functions: knowledge elicitation, knowledge representation, and cross-functional integration. We illustrated the utility of this approach by applying it to practical problems and case studies. The complete report is available in [1]; this paper is a description of our approach and report contents.

99 GENERAL AND MISCELLANEOUS↗

Human-building interaction at work: Findings from an interdisciplinary cross-country survey in Italy

This study presents results from an interdisciplinary survey assessing contextual and behavioral factors driving occupants’ interaction with building and systems in offices located across three different Mediterranean climates in Turin (Northern), Perugia (Central), and Rende (Southern) Italy. The survey instrument is grounded in an interdisciplinary framework that bridges the gap between building physics and social science environments on the energy- and comfort-related human-building interaction in the workspace. Outcomes of the survey questionnaire provide insights into four key learning objectives: (1) individual occupant's motivational drivers regarding interaction with shared building environmental controls (such as adjustable thermostats, operable windows, blinds and shades, and artificial lighting), (2) group dynamics such as perceived social norms, attitudes, and intention to share controls, (3) occupant perception of the ease of use and knowledge of how to operate control systems, and (4) occupant-perceived comfort, satisfaction, and productivity. The study attempts to identify climatic, cultural, and socio-demographic influencing factors, as well as to establish the validity of the survey instrument and robustness of outcomes for future studies. Also, the paper aims at illustrating why and how social science insights can bring innovative knowledge into the adoption of building technologies in shared contexts, thus enhancing perceived environmental satisfaction and effectiveness of personal indoor climate control in office settings and impacting office workers’ productivity and reduced operational energy costs.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Data-driven Minimum Entropy Control for Stochastic Nonlinear Systems using the Cumulant-Generating Function

Here, we present a novel minimum entropy control algorithm for a class of stochastic nonlinear systems subjected to non-Gaussian noises. The entropy control can be considered as an optimization problem for the system randomness attenuation, but the mean value has to be considered separately. To overcome this disadvantage, a new representation of the system stochastic properties was given using the cumulant-generating function based on the moment-generating function, in which the mean value and the entropy was reflected by the shape of the cumulant-generating function. Based on the samples of the system output and control input, a time-variant linear model was identified, and the minimum entropy optimization was transformed to system stabilization. Then, an optimal control strategy was developed to achieve the randomness attenuation, and the boundedness of the controlled system output was analyzed. The effectiveness of the presented control algorithm was demonstrated by a numerical example. In this paper, a data-driven minimum entropy design is presented without pre-knowledge of the system model; entropy optimization is achieved by the system stabilization approach in which the stochastic distribution control and minimum entropy are unified using the same identified structure; and a potential framework is obtained since all the existing system stabilization methods can be adopted to achieve the minimum entropy objective.

42 ENGINEERING↗

Industrial Energy Training and Certification

Compressed air systems, ammonia refrigeration systems, chilled water systems, steam systems, process heating systems, combined heat and power systems, pump systems and fan systems are major industrial energy systems commonly found in manufacturing facilities. Efficiency of these systems contributes significantly to whole facilities' energy performance. On the national, even international level, well-structured training and highly recognized certification programs help develop a highly-skilled and qualified workforce to maintain and improve facilities' energy performance, particularly as technologies within these systems become more advanced. The purpose of this paper is to review currently available training and certification programs focusing on these systems and to identify the gap between market's needs and currently available programs. Three major conclusions are: first, most training programs focus on operations, maintenance, safety and design although some briefly touch the energy performance aspect; second, except CRES by RETA and PSA Certificate and PSAP Master Certification by HI, no other certifications had been found emphasizing on knowledge and skills for improving and maintaining these systems' energy performance; third, developing energy efficiency focused training and ANSI accredited certification programs on these energy systems will fill the gap between market's needs and currently available programs.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Industrial Energy Training and Certification

We present compressed air systems, ammonia refrigeration systems, chilled water systems, steam systems, process heating systems, combined heat and power systems, pump systems and fan systems are major industrial energy systems commonly found in manufacturing facilities. Efficiency of these systems contributes significantly to whole facilities' energy performance. On the national, even international level, well-structured training and highly recognized certification programs help develop a highly-skilled and qualified workforce to maintain and improve facilities' energy performance, particularly as technologies within these systems become more advanced. The purpose of this paper is to review currently available training and certification programs focusing on these systems and to identify the gap between market's needs and currently available programs. Three major conclusions are: first, most training programs focus on operations, maintenance, safety and design although some briefly touch the energy performance aspect; second, except CRES by RETA and PSA Certificate and PSAP Master Certification by HI, no other certifications had been found emphasizing on knowledge and skills for improving and maintaining these systems' energy performance; third, developing energy efficiency focused training and ANSI accredited certification programs on these energy systems will fill the gap between market's needs and currently available programs.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Approaches for handling high-dimensional cluster expansions of ionic systems

Disordered multicomponent systems attract great interest due to their engineering design flexibility and subsequent rich space of properties. However, detailed characterization of the structure and atomic correlations remains challenging and hinders full navigation of these complex spaces. A lattice cluster expansion is one tool to obtain configurational and energetic resolution. While in theory a cluster expansion can be applied to any system of any dimensionality, the method has primarily been used in binary systems or ternary alloys. Here we apply cluster expansions in high-component ionic systems, setting up the largest cluster expansion ever attempted to our knowledge. In doing so, we address and discuss challenges specific to high-component ionic systems, namely charge state assignments, structural relaxations, and rank-deficient systems. We introduce practical procedures to make the fitting and analysis of complex systems tractable, providing guidance for future computational studies of disordered ionic systems.

25 ENERGY STORAGE↗

Performance contracting centers of expertise: a framework for federal implementation

This report examines the establishment and operation of Energy Performance Contracting (EPC) Centers of Expertise (COEs) within the federal government. EPCs, including Energy Savings Performance Contracts (ESPCs) and Utility Energy Service Contracts (UESCs), are critical mechanisms for advancing energy efficiency, resilience, and infrastructure modernization without the need for significant upfront appropriations. However, EPCs require specialized knowledge in project development, contracting, financing, legal parameters and technical project oversight. Currently federal agencies have varying levels of expertise and institutionalized policy to effectively and consistently use congressionally authorized EPCs which have decades of proven and impactful use. To address these challenges, several federal agencies have created COEs to centralize expertise, standardize practices, and streamline implementation. This report reviews statutory and policy drivers, highlights the benefits and challenges and presents case studies from the General Services Administration (GSA), the Department of Veterans Affairs (VA) and the U.S. Army Engineering and Support Center Huntsville (HNC). Recommendations are also provided for agencies considering the establishment of EPC COEs, which will bring much needed structure, consistency and lead to implementation of these energy and infrastructure building projects to save costs for U.S. taxpayers.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗