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At least 145 records · Page 8
All sourcesNASA NTRSDOE OSTI
DOE OSTI2020

A global lipid map defines a network essential for Zika virus replication

>Zika virus (ZIKV), an arbovirus of global concern, remodels intracellular membranes to form replication sites. How ZIKV dysregulates lipid networks to allow this, and consequences for disease, is poorly understood. Here, we performed comprehensive lipidomics to create a lipid network map during ZIKV infection. We found that ZIKV significantly alters host lipid composition, with the most striking changes seen within subclasses of sphingolipids. Ectopic expression of ZIKV NS4B protein resulted in similar changes, demonstrating a role for NS4B in modulating sphingolipid pathways. Disruption of sphingolipid biosynthesis in various cell types, including human neural progenitor cells, blocked ZIKV infection. Additionally, the sphingolipid ceramide redistributes to ZIKV replication sites and increasing ceramide levels by multiple pathways sensitizes cells to ZIKV infection. Thus, we identify a sphingolipid metabolic network with a critical role in ZIKV replication and show that ceramide flux is a key mediator of ZIKV infection.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2026

Artificial Soiling Replication of Field Losses on Commercial Photovoltaic Modules

Here, this study demonstrates the capabilities of an indoor artificial soiling approach developed to closely replicate the natural, cyclic soil accumulation processes in the field-dust suspension, deposition, and sedimentation/cementation-for a subtropical climate. In this work, a near-field environment is replicated in an artificial soiling cubic chamber through controlled regulation of humidity, temperature, dust type, and dust concentration, based on site-specific historical climate data. Two different models (MA and MB) of full-size commercial photovoltaic modules from a single manufacturer, installed side by side in the mid-Atlantic United States, were retrieved and subjected to artificial soiling experiments and various characterization measurements, including short-circuit current, colorimetry, reflectance, X-ray fluorescence, laser diffraction, and optical microscopy. Both in the field and in our improved field-representative artificial soiling tests, the MA model experienced roughly twice the soiling loss as the MB model. To closely replicate the field soiling losses for a site-specific climate, it is critical to include: 1) The use of field-collected dust with identical dust chemistry and particle distribution instead of standardized ISO 12103 Arizona Road dusts, 2) the use of only a small amount of field-collected dust inside the chamber during the deposition process (e.g., 0.15 g), and 3) the preconditioning of the surface coating for the partial/full dose of UV stress as experienced in the field during sunlight exposure and the abrasion as experienced in the field during regular module cleaning activities, if/as needed. The field-representative artificial soiling method developed here could potentially be adopted for rank ordering of various antisoiling coatings developed by researchers and industry stakeholders.

14 SOLAR ENERGY↗
DOE OSTI2021

Processing Aleatory and Epistemic Uncertainties in Experimental Data From Sparse Replicate Tests of Stochastic Systems for Real-Space Model Validation

This paper presents a practical methodology for propagating and processing uncertainties associated with random measurement and estimation errors (that vary from test-to-test) and systematic measurement and estimation errors (uncertain but similar from test-to-test) in inputs and outputs of replicate tests to characterize response variability of stochastically varying test units. Also treated are test condition control variability from test-to-test and sampling uncertainty due to limited numbers of replicate tests. These aleatory variabilities and epistemic uncertainties result in uncertainty on computed statistics of output response quantities. The methodology was developed in the context of processing experimental data for “real-space” (RS) model validation comparisons against model-predicted statistics and uncertainty thereof. The methodology is flexible and sufficient for many types of experimental and data uncertainty, offering the most extensive data uncertainty quantification (UQ) treatment of any model validation method the authors are aware of. It handles both interval and probabilistic uncertainty descriptions and can be performed with relatively little computational cost through use of simple and effective dimension- and order-adaptive polynomial response surfaces in a Monte Carlo (MC) uncertainty propagation approach. A key feature of the progressively upgraded response surfaces is that they enable estimation of propagation error contributed by the surrogate model. Sensitivity analysis of the relative contributions of the various uncertainty sources to the total uncertainty of statistical estimates is also presented. Finally, the methodologies are demonstrated on real experimental validation data involving all the mentioned sources and types of error and uncertainty in five replicate tests of pressure vessels heated and pressurized to failure. Simple spreadsheet procedures are used for all processing operations.

97 MATHEMATICS AND COMPUTING↗
DOE OSTI2022

Fabrication of low blaze angle gratings by replication and plasma etch

We suggest a new method of making ultra-low blaze angle gratings for synchrotron application. The method is based on reduction of the blaze angle of a master grating by replication followed by a plasma etch. A master blazed grating with a relatively large blaze angle is fabricated by anisotropic wet etching of a Si single crystal substrate. The surface of the master grating is replicated by a polymer material on top of a quartz substrate by nanoimprinting and then transferred into quartz by a plasma etch. Then a 2nd nanoimprint step is applied to transfer the saw-tooth surface into a resist layer on top of a Si grating substrate. The plasma etch through the patterned resist layer provides transfer of the grooves into the Si substrate and results in reduction of the blaze angle due to the difference in etch rates of the resist and Si. We investigated the impact of the replication process on the groove shape, facet surface roughness, and diffraction efficiency of the fabricated 200 lines/mm low blaze angle grating.

Park, Sooyeon↗
DOE OSTI2023

Influenza A Virus Multicycle Replication Yields Comparable Viral Population Emergence in Human Respiratory and Ocular Cell Types

While primarily considered a respiratory pathogen, influenza A virus (IAV) is nonetheless capable of spreading to, and replicating in, numerous extrapulmonary tissues in humans. However, within-host assessments of genetic diversity during multicycle replication have been largely limited to respiratory tract tissues and specimens. As selective pressures can vary greatly between anatomical sites, there is a need to examine how measures of viral diversity may vary between influenza viruses exhibiting different tropisms in humans, as well as following influenza virus infection of cells derived from different organ systems. Here, we employed human primary tissue constructs emulative of the human airway or corneal surface, and we infected both with a panel of human- and avian-origin IAV, inclusive of H1 and H3 subtype human viruses and highly pathogenic H5 and H7 subtype viruses, which are associated with both respiratory disease and conjunctivitis following human infection. While both cell types supported productive replication of all viruses, airway-derived tissue constructs elicited greater induction of genes associated with antiviral responses than did corneal-derived constructs. We used next-generation sequencing to examine viral mutations and population diversity, utilizing several metrics. With few exceptions, generally comparable measures of viral diversity and mutational frequency were detected following homologous virus infection of both respiratory-origin and ocular-origin tissue constructs. Expansion of within-host assessments of genetic diversity to include IAV with atypical clinical presentations in humans or in extrapulmonary cell types can provide greater insight into understanding those features most prone to modulation in the context of viral tropism.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Advanced low blaze angle x-ray gratings via nanoimprint replication and plasma etch

We developed a new method of making ultra-low blaze angle diffraction gratings for x-ray applications. The method is based on reduction of the blaze angle of a master grating by nanoimprint replication followed by a plasma etch. A master blazed grating with a relatively large blaze angle is fabricated by anisotropic wet etching of a Si single crystal substrate. The surface of the master grating is replicated by a polymer material on top of a quartz substrate by nanoimprinting. Then a second nanoimprinting is performed using the 1st replica as a mold to replicate the saw-tooth surface into a resist layer on top of a Si grating substrate. A reactive ion etch is used to transfer the grating grooves into the Si substrate. The plasma etch provides reduction of the groove depth by a factor defined by the ratio of the etch rates for the resist and Si. We demonstrate reduction of the blaze angle of a master grating by a factor of 5 during fabrication of a 200 lines/mm blazed grating with a blaze angle of 0.2°. We investigated the quality and performance of the fabricated low blaze angle gratings and evaluate process accuracy and reproducibility. The new blaze angle reduction method preserves the planarity of the optical surface of the grating substrate and at the same time provides improvement in the grating groove quality during the reduction process.

47 OTHER INSTRUMENTATION↗
DOE OSTI2021

Evaluation of the Function of the ASFV KP177R Gene, Encoding for Structural Protein p22, in the Process of Virus Replication and in Swine Virulence

African swine fever virus (ASFV) causes a devastating disease of swine that has caused outbreaks in Central Europe since 2007, spreading into Asia in 2018. ASFV is a large, structurally complex virus with a large dsDNA genome encoding for more than 160 genes, most of them still uncharacterized. p22, encoded by the ASFV gene KP177R, is an early transcribed, structural virus protein located in the ASFV particle. Although its exact function is unknown, p22 has recently been identified as an interacting partner of several host proteins. Here, we describe the development of a recombinant ASFV (ASFV-G-ΔKP177R) lacking the KP177R gene as a tool to evaluate the role of p22 in virus replication and virulence in swine. The recombinant ASFV-G-ΔKP177R demonstrated that the KP177R gene is non-essential for ASFV replication in primary swine macrophages, with virus yields similar to those of the parental, highly virulent field isolate Georgia2010 (ASFV-G). In addition, experimental infection of domestic pigs with ASFV-G-ΔKP177R produced a clinical disease similar to that caused by the parental ASFV-G. Therefore, and surprisingly, p22 does not seem to be involved in virus replication or virulence in swine.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Evaluation of the Function of ASFV Gene E66L in the Process of Virus Replication and Virulence in Swine

African swine fever virus (ASFV) is the etiological agent of an economically important disease of swine currently affecting large areas of Africa, Eurasia and the Caribbean. ASFV has a complex structure harboring a large dsDNA genome which encodes for more than 160 proteins. One of the proteins, E66L, has recently been involved in arresting gene transcription in the infected host cell. Here, we investigate the role of E66L in the processes of virus replication in swine macrophages and disease production in domestic swine. A recombinant ASFV was developed (ASFV-G-ΔE66L), from the virulent parental Georgia 2010 isolate (ASFV-G), harboring the deletion of the E66L gene as a tool to assess the role of the gene. ASFV-G-ΔE66L showed that the E66L gene is non-essential for ASFV replication in primary swine macrophages when compared with the parental highly virulent field isolate ASFV-G. Additionally, domestic pigs infected with ASFV-G-ΔE66L developed a clinical disease undistinguishable from that produced by ASFV-G. Therefore, E66L is not involved in virus replication or virulence in domestic pigs.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Assessment of the Effect of Deleting the African Swine Fever Virus Gene R298L on Virus Replication and Virulence of the Georgia2010 Isolate

African swine fever (ASF) is a lethal disease of domestic pigs that is currently challenging swine production in large areas of Eurasia. The causative agent, ASF virus (ASFV), is a large, double-stranded and structurally complex virus. The ASFV genome encodes for more than 160 proteins; however, the functions of most of these proteins are still in the process of being characterized. The ASF gene R298L, which has previously been characterized as able to encode a functional serine protein kinase, is expressed late in the virus infection cycle and may be part of the virus particle. There is no description of the importance of the R298L gene in basic virus functions such as replication or virulence in the natural host. Based on its evolution, it is proposed that there are four different phenotypes of R298L of ASFV in nature, which may have potential implications for R298L functionality. We report here that a recombinant virus lacking the R298L gene in the Georgia 2010 isolate, ASFV-G-∆R298L, does not exhibit significant changes in its replication in primary cultures of swine macrophages. In addition, when experimentally inoculated in pigs, ASFV-G-∆R298L induced a fatal form of the disease similar to that caused by the parental virulent ASFV-G. Therefore, deletion of R298L does not significantly affect virus replication and virulence in domestic pigs of the ASFV Georgia 2010 isolate.

Virology↗
DOE OSTI2025

Vaccine Efficacy of a Replication-Competent Interferon-Expressing Porcine Reproductive and Respiratory Syndrome (PRRS) Virus Against NADC-34 Challenge

Background/Objectives: Porcine reproductive and respiratory syndrome virus (PRRSV) significantly impedes swine production due to rapid genetic variation and suppression of antiviral interferon (IFN) responses, leading to ineffective immunity. To address this, we developed IFNmix, a replication-competent PRRSV modified live vaccine (MLV) candidate co-expressing three Type I IFN subclasses (IFNα, IFNβ, IFNδ) to enhance antiviral immunity. Methods: In two independent in vivo experiments, we compared the protection of IFNmix and a commercial PRRSV MLV vaccine during challenge with a virulent PRRSV strain. Clinical signs, antibody and cytokine production, viral replication, and lung pathology in IFNmix-vaccinated pigs were compared to those of commercial PRRSV vaccines and controls. Results: Pigs vaccinated with IFNmix exhibited similar anti-PRRSV antibody development, serum viral loads, lung lesions, and cytokine responses post-challenge with the virulent NADC34 strain, with comparable or lower body temperatures and weight gain, to pigs vaccinated with the commercial vaccines. While IFNmix showed early viral load reduction compared to the commercial vaccine (Days 7–14 post-challenge), it demonstrated similar efficacy in controlling PRRSV replication and lung pathology. Conclusions: These findings suggest that IFNmix, by expressing multiple IFNs, can potentially enhance innate and adaptive immune responses, offering a promising approach to improving PRRSV vaccine efficacy. Further studies are needed to evaluate IFNmix against a broader range of PRRSV strains and to optimize its attenuation and immunogenicity.

Miller, Laura C. (ORCID:0000000289469416)↗
NASA NTRS1974

Prebiotic chemistry and nucleic acid replication

Recent work is reviewed on some reactions that could have occurred on the primitive earth and that could have played a part in the evolution of a self-replicating system. The transition from the primitive atmosphere to the simplest replicating molecules is considered in four stages: (1) the formation of a 'prebiotic soup' of organic precursors, including the purine and pyrimidine bases and the pentose sugars; (2) the condensation of these precursors and inorganic phosphate to form monomeric nucleotides and activated nucleotide derivatives; (3) the polymerization of nucleotide derivatives to oligonucleotides; and (4) the complementary replication of oligonucleotides in a template-directed process that depends on Watson-Crick base pairing.

Orgel, L. E.↗
NASA NTRS1982

Management and control of self-replicating systems: A systems model

In 1980, a conceptual engineering approach to self-replicating systems was achieved. The design was based on von Newmann's kinematic version of self-replicating automata. The systems management and control and the organization of the control elements are reported. After developing the functional requirements of such a system, a hierarchy of three management and control levels is described. These are an autonomous, an external, and an intelligent management and control system. Systems recycling, systems specialization, and information replication are discussed.

Vontiesenhausen, G.↗
NASA NTRS1992

Molecular replication

Recent experiments demonstrating nonenzymatic replication in molecular systems are reviewed. The difficulties facing nonenzymatic replication are discussed along with specificity, fidelity, and mutation in nonenzymatic replication. The prospects for research in this area are considered.

Orgel, Leslie E.↗
NASA NTRS1993

Investigate zero-stress replicated optics

The contracted activities for the procurement of 'Investigate Zero-Stress Replicated Optics' to support the AXAF-S x-ray spectrometer mirrors has been completed. To date four large Wolter I grazing incidence x-ray optical shells have been electroformed from nickel. The mirrors were fabricated utilizing each of two nickel alloy plated aluminum substrates twice. A wide variety of testing has been completed by NASA MSFC and UAH. This testing includes heat treatment control tests, subscale plating and fixture testing, alloy control of the electroless nickel, adhesion and release testing of the gold to electroless nickel, electroforming instrumentation and software and fabrication of subscale models. The full scale shells are one millimeter thick nickel electrodeposited over a thin gold layer which in turn has the optical surface on the inside. The optical surface is the replicate of the surface prepared on the substrate. Appendix I briefly outlines the fabrication process. Major objectives which were shared by UAH and MSFC include the design of facilities, equipment and tooling and procurement of materials and equipment. Process development followed with the fabrication of small scale pilot units. Procurement commenced immediately and equipment and materials were ordered to implement the fabrication of first surface full scale substrates (mandrels) and the second surface electroformed optical components. All principal objectives have been achieved. Inspection of the mirrors in visible and x-ray modes validates that the required performance and the quality can be achieved by an electroforming replication process. A very distinct progressive improvement has been achieved with each of the four mirrors produced. The final mirror exceeded the original goals and set an improved standard for flight hardware. The future goal of a 30 arc second resolution at 8 KEV x-ray appears to be achievable by this process when proper cleanliness and process control is utilized.

Engelhaupt, Darell↗
NASA NTRS1994

A novel computational method to simulate non-enzymatic self-replication

Non-enzymatic, template-directed synthesis of oligonucleotides has been extensively studied in the laboratory as a model to understand the kind of chemical processes that might have contributed to the origin of life on Earth. Several oligonucleotides have been shown to catalyze the synthesis of their complements from activated mononucleotides; however, a restricted number of them have been found to self-replicate. Recently we developed an efficient modified cellular automata method that supports the study of self-replicating oligonucleotides. With this method the oligonucleotide molecules are represented as active cells imbedded in a two-dimensional array of inactive cells symbolizing the environment. Random movements and probability-governed chemical reactions occurring in a cellular space can effectively simulate the experimental behavior observed in self-directed replication of oligonucleotides.

Navarro-Gonzalez, Rafael↗
NASA NTRS1995

Development of replicated optics for AXAF-1 XDA testing

Advanced optical systems for applications such as grazing incidence Wolter I x-ray mirror assemblies require extraordinary mirror surfaces in terms of fine finish and surface figure. The impeccable mirror surface is on the inside of the rotational mirror form. One practical method of producing devices with these requirements is to first fabricate an exterior surface for the optical device then replicate that surface to have the inverse component with lightweight characteristics. The replicated optic is not better than the master or mandrel from which it is made. This task identifies methods and materials for forming these extremely low roughness optical components. The objectives of this contract were to (1) prepare replication samples of electroless nickel coated aluminum, and determine process requirements for plating XDA test optic; (2) prepare and assemble plating equipment required to process a demonstration optic; (3) characterize mandrels, replicas and test samples for residual stress, surface contamination and surface roughness and figure using equipment at MSFC and; (4) provide technical expertise in establishing the processes, procedures, supplies and equipment needed to process the XDA test optics.

Engelhaupt, Darell↗
NASA NTRS1996

A Flexible Alignment Fixture for the Fabrication of Replication Mandrels

NASA uses precision diamond turning technology to fabricate replication mandrels for its X-ray Calibration Facility (XRCF) optics. The XRCF optics are tubular, and the internal surface contains a parabolic profile over the first section and a hyperbolic profile over the last. The optic is fabricated by depositing layers of gold and nickel on to the replication mandrel and then separating it from the mandrel. Since the mandrel serves as a replication form, it must contain the inverse image of the surface. The difficulty in aligning the mandrel comes from the fabrication steps which it undergoes. The mandrel is rough machined and heat treated prior to diamond turning. After diamond turning, silicon rubber separators which are undercut in radius by 3 mm (0.12 in.) are inserted between the two end caps of the mandrel to allow the plating to wrap around the ends (to prevent flaking). The mandrel is then plated with a nickel-phosphor alloy using an electroless nickel process. At this point, the separators are removed and the mandrel is reassembled for the final cut on the DTM. The mandrel is measured for profile and finish, and polished to achieve an acceptable surface finish. Wrapping the plating around the edges helps to prevent flaking, but it also destroys the alignment surfaces between the parts of the mandrel that insure that the axes of the parts are coincident. Several mandrels have been realigned by trial-and-error methods, consuming significant amounts of setup time. When the mandrel studied in this paper was reassembled, multiple efforts resulted in a minimum radial error motion of 100 microns. Since 50 microns of nickel plating was to be removed, and a minimum plating thickness of 25 microns was to remain on the part, the radial error motion had to be reduced to less than 25 microns. The mandrel was therefore not usable in its current state.

Cuttino, James F.↗
NASA NTRS1999

Superpolishing and Precision Metrology on a Metal Mandrel and Replicated Segments for Constellation-X

We have superpolished a diamond-turned aluminum mandrel (coated with electroless Ni) to an axial roughness of 6.34 nm rms. The mandrel is made to the Astro-E secondary mirror design for the 81st shell. Precision metrology at 100 mm to submicron scales has established the power spectral density of the mandrel and ultralightweight gold coated replicated segments. Predicted image quality of a set of optimally aligned replicated segments of this and a matching primary is substantially improved as compared to the flight mirrors for Astro-E. This approach using metal mandrels, superpolishing, and replicated ultralightweight foil mirrors, may represent a cost-effective approach to meeting the 15 arcsec half-energy width and weight requirements for the Constellation-X mission. Descriptions of the polishing apparatus, the precision metrology instruments, and the surface data analysis are presented. The general methods described are applicable to precision optics for both normal incidence and grazing incidence optics.

Content, D.↗