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Computer simulations of the experiments are being performed using the Los Alamos National Laboratories’ FLAG magnetohydrodynamics code by graduate student Seth Kreher, and Lawrence Livermore National Laboratory’s Ares by graduate students Robert Masti and Matthew Carrier.
Abstract not provided.
The widespread Pseudomonas genus comprises a collection of related species with remarkable abilities to degrade plastics and polluted wastes and to produce a broad set of valuable compounds, ranging from bulk chemicals to pharmaceuticals. Pseudomonas possess characteristics of tolerance and stress resistance making them valuable hosts for industrial and environmental biotechnology. However, efficient and high-throughput genetic engineering tools have limited metabolic engineering efforts and applications. To improve their genome editing capabilities, we first employed a computational biology workflow to generate a genus-specific library of potential single-stranded DNA-annealing proteins (SSAPs). Assessment of the library was performed in different Pseudomonas using a high-throughput pooled recombinase screen followed by Oxford Nanopore NGS analysis. Among different active variants with variable levels of allelic replacement frequency (ARF), efficient SSAPs were found and characterized for mediating recombineering in the four tested species. New variants yielded higher ARFs than existing ones in Pseudomonas putida and Pseudomonas aeruginosa, and expanded the field of recombineering in Pseudomonas taiwanensisand Pseudomonas fluorescens. These findings will enhance the mutagenesis capabilities of these members of the Pseudomonas genus, increasing the possibilities for biotransformation and enhancing their potential for synthetic biology applications.
Slides to be presented during the NNSA 2021 Stewardship Science Academic Programs (SSAP) Symposium (https://web.cvent.com/event/1893b742-2d98-49d5-87cc-f07b1c3cb66c/summary?environment=P2). The symposium will be held virtually February 16–18, 2021.
I am a staff scientist at Sandia National Laboratories (SNL). I work on multiple fundamental-science projects and lead modeling/data-analysis for the stellar opacity experiments. I also work with theorists to refine plasma material-property calculations to make NNSA simulations more predictable. These are challenging and important problems for national security. In fact, our experiments raised questions about “opacity”, the property of matter that controls energy transport inside stars. This work was published by Nature due to its serious implication over broad applications, and our team received NNSA Defense Program Award of Excellence, both in 2015. Work at SNL is collegial and full of learning through interactions with the world’s finest experimentalists and theorists. When I started my Ph.D. program, working at a national lab was a dream position. Today, I collaborate with top scientists to solve mission-critical problems. The Stewardship Science Academic Programs (SSAP) and National Laser Users’ Facility (NLUF) played a vital role in the efficient learning and seamless transition to SNL.
Presentation for the NNSA's 2022 Stewardship Science Academic Programs (SSAP) Symposium February 15-17, 2022
This document summarizes the activities at the Mykonos Pulsed Power Facility during the calendar year 2025. Section 1 reports on the yearly shot statistics along with some facility highlights. Section 2 discusses the many improvements we were able to complete this year, thanks to the generous Mykonos Academic Access Programs (MAAP) funding from SSAP and Sandia strategic investments. Section 3 focuses on each individual campaign and their respective results.
Outlined here is a preliminary design for a Solar Power Satellite (SPS) system. The SPS will provide a clean, reliable source of energy for mass consumption. The system will use satellites in geostationary orbits around the Earth to capture the sun's energy. The intercepted sunlight will be converted to laser beam energy which can be transmitted to the Earth's surface. Ground systems on the Earth will convert the transmissions from space into electric power. The preliminary design for the SPS consists of one satellite in orbit around the Earth transmitting to one ground station. The SPs technology uses multi-layer solar cell technology arranged on a 20 sq km planar array to intercept sunlight and convert it to an electric voltage. Power conditioning devices then send the electricity to a laser, which transmits the power to the surface of the Earth. A ground station will convert the beam into electricity. Construction will take place in low Earth orbit and array sections, 20 in total, will be sailed on the solar wind out to the GEO location in 150 days. These individual transportation sections are referred to as solar sailing panels (SSAPs). The primary truss elements used to support the arrays are composed on composite tubular members in a pentahedral arrangement. Smart segments consisting of passive and active damping devices will increase the control of dynamic SPS modes.
The solar power satellite (SPS) will provide a clean, reliable source of energy for large-scale consumption. The system will use satellites in geostationary orbits around the Earth to capture the Sun's energy. The intercepted sunlight will be converted to laser beam energy that can be transmitted to the Earth's surface. Ground systems on the Earth will convert the transmissions from space into electric power. The preliminary design for the SPS consists of one satellite in orbit around the Earth transmitting energy to a single ground station. The SPS design uses multilayer solar cell technology arranged on a 20 km squared planar array to intercept sunlight and convert it to an electric voltage. Power conditioning devices then send the electricity to a laser, which transmits the power to the surface of the Earth. A ground station will convert the beam into electricity. Typically, a single SPS will supply 5 GW of power to the ground station. Due to the large mass of the SPS, about 41 million kg, construction in space is needed in order to keep the structural mass low. The orbit configuration for this design is to operate a single satellite in geosynchronous orbit (GEO). The GEO allows the system to be positioned above a single receiving station and remain in sunlight 99 percent of the time. Construction will take place in low Earth orbit (LEO); array sections, 20 in total, will be sailed on solar wind out to the GEO location in 150 days. These individual transportation sections are referred to as solar sailing array panels (SSAP's). The primary truss elements used to support the array are composed of composite tubular members in a pentahedral arrangement. Smart segments consisting of passive and active damping devices will increase the control of dynamic SPS modes.
Returning samples from Mars has long been a major planetary science objective due to the high scientific value and transformative potential of the resulting data. An exciting dimension of this objective is the potential for the detection of ancient microbiological life, and the possibility of improving our understanding of the evolution of habitable environments on Mars and the development of life on Earth. To ensure that returned samples meet stringent planetary protection requirements and do not expose Earth to potential biohazards, the joint NASA/ESA Sample Receiving Project (SRP) assembled the Sample Safety Assessment Protocol Tiger Team (SSAP-TT). Members were recruited with the specific goal of creating a multi-disciplinary and internationally distributed team of experts in their respective fields across the federal government, academia, and private industry. This team was chartered with reassessing previous sample safety assessment strategies, defining what constitutes a biological hazard, developing a protocol to test for potential biohazards, and establishing a statistical framework to determine if samples are “safe” for release. The team developed a three-step protocol, supported by a Bayesian statistical framework, to assess whether returned samples contain potential biohazards that could present a risk to Earth’s biosphere. Initial conclusions indicated that an effective and comprehensive safety assessment protocol is feasible using modern techniques and does not require an excessive amount of sample consumption or traditional microbiological detection methodology. Herein, we will present an overview of the MSR SRP, the proposed safety assessment protocol, and how aspects of this novel approach can be applied to biological assessment in healthcare product manufacturing practices.
ABSTRACT Several strains in the Pseudomonas genus are categorized as plant growth-promoting rhizobacteria (PGPR). Although several of these strains are strong candidates for applications as biofertilizers or biopesticides, genome editing approaches are generally limited and require further development. Editing genomes in PGPR could enable more robust agricultural applications, persistence, and biosafety measures. In this study, we investigate the use of five phage-encoded recombinases to develop a recombineering workflow in three PGPR strains: Pseudomonas protegens Pf-5, Pseudomonas protegens CHA0, and Pseudomonas putida KT2440. Using point mutations in the rpoB gene, we reach maximum recombineering efficiencies of 1.5 × 10 −4 , 3 × 10 −4 , and 5 × 10 −5 , respectively, in these strains using λ-Red Beta recombinase from Escherichia coli . We further examine recombineering efficiencies across these strains as a function of selected mutation, editing template concentration, and phosphorothiolate bond protection. This work validates the use of these tools across several environmentally and biotechnologically relevant strains to expand the possibilities of genetic manipulation in the Pseudomonas genus. IMPORTANCE The Pseudomonas genus contains many members currently being investigated for applications in biodegradation, biopesticides, biocontrol, and synthetic biology. Though several strains have been identified with beneficial properties, chromosomal manipulations to further improve these strains for commercial applications have been limited due to the lack of efficient genetic tools that have been tested across this genus. Here, we test the recombineering efficiencies of five phage-derived recombinases across three biotechnologically relevant Pseudomonas strains: P. putida KT2440, P. protegens Pf-5, and P. protegens CHA0. These results demonstrate a method to generate targeted mutations quickly and efficiently across these strains, ideally introducing a method that can be implemented across the Pseudomonas genus and a strategy that may be applied to develop analogous systems in other nonmodel bacteria.