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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 55 records · Page 3
All sourcesNASA NTRSDOE OSTI
DOE OSTI2021

Spatially resolved photoluminescence brightening in individual single-walled carbon nanotubes

Single-molecule microscopy was used to image photoluminescence (PL) brightening of individual sodium-dodecyl-sulfate (SDS)-wrapped single-walled carbon nanotubes (SWCNTs) upon the addition of dithiothreitol (DTT). PL enhancement varied for each nanotube (NT), with some brightening by 16% and others by a factor of about 7. Interestingly, NTs that displayed lower initial QY values showed the largest increases in PL enhancement. SDS-SWCNTs longer than the diffraction limit were studied in order to spatially resolve the brightening phenomenon. Quite unexpectedly, a uniform, single-step PL brightening along the NT was consistently observed, suggesting that the PL enhancement is the result of a non-localized process. Furthermore, the even PL brightening seen over SWCNTs that are micrometers long implies that single point defect sites, which are known to be largely responsible for exciton nonradiative decay, play no significant role in the brightening process. Interestingly, affixing the SWCNT strongly to the substrate surface mitigated the PL brightening response, consistent with a hypothesis that surfactant reorganization upon the addition of DTT is responsible for exciton PL brightening.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗
DOE OSTI2022

Growing impact of wildfire on western US water supply

Streamflow often increases after fire, but the persistence of this effect and its importance to present and future regional water resources are unclear. This paper addresses these knowledge gaps for the western United States (WUS), where annual forest fire area increased by more than 1,100% during 1984 to 2020. Among 72 forested basins across the WUS that burned between 1984 and 2019, the multibasin mean streamflow was significantly elevated by 0.19 SDs (P < 0.01) for an average of 6 water years postfire, compared to the range of results expected from climate alone. Significance is assessed by comparing prefire and postfire streamflow responses to climate and also to streamflow among 107 control basins that experienced little to no wildfire during the study period. The streamflow response scales with fire extent: among the 29 basins where >20% of forest area burned in a year, streamflow over the first 6 water years postfire increased by a multibasin average of 0.38 SDs, or 30%. Postfire streamflow increases were significant in all four seasons. Historical fire–climate relationships combined with climate model projections suggest that 2021 to 2050 will see repeated years when climate is more fire-conducive than in 2020, the year currently holding the modern record for WUS forest area burned. These findings center on relatively small, minimally managed basins, but our results suggest that burned areas will grow enough over the next 3 decades to enhance streamflow at regional scales. Wildfire is an emerging driver of runoff change that will increasingly alter climate impacts on water supplies and runoff-related risks.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Increased radiation due to non-coronal effects on DIII-D and MAST-U with varying input power

Through SOLPS-ITER simulations of DIII-D and MAST-U, an X-divertor (XD) on DIII-D and a super X-divertor (SXD) on MAST-U were shown to have increased carbon emissivity (P Rad /n e n I ) over corresponding standard divertors (SD) at similar degrees of partial detachment. The reasons behind the increased emissivity in the DIII-D XD and SXD are analyzed using a simple 0D transport model. From the transport model, it is seen that a major cause of the increased emissivity in the XD and SXD over the SDs is a shorter impurity confinement time. An additional cause (for the SXD) is an increase in the ratio of neutral hydrogen to electron density. The input power (P in ) was varied and the XD had a higher emissivity at the higher P in , unlike the SDs which had the emissivity decrease with increasing P in . A basic geometrical reason is given to explain both the benefits of the XD over the SD as well as the increase in the XD's emissivity with P in .

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗
DOE OSTI2021

Purification of native histidine-rich protein 2 (nHRP2) from Plasmodium falciparum culture supernatant, infected RBCs, and parasite lysate

Background: Despite the widespread use of histidine-rich protein 2 (HRP2)-based rapid diagnostic tests (RDTs), purified native HRP2 antigen is not standardly used in research applications or assessment of RDTs used in the field. Methods: This report describes the purification of native HRP2 (nHRP2) from the HB3 Plasmodium falciparum culture strain. As this culture strain lacks pfhrp3 from its genome, it is an excellent source of HRP2 protein only and does not produce the closely-related HRP3. The nHRP2 protein was isolated from culture supernatant, infected red blood cells (iRBCs), and whole parasite lysate using nickel-metal chelate chromatography. Biochemical characterization of nHRP2 from HB3 culture was conducted by SDS-PAGE and western blotting, and nHRP2 was assayed by RDT, ELISA, and bead-based immunoassay. Results: Purified nHRP2 was identified by SDS-PAGE and western blot as a –60 kDa protein that bound anti-HRP-2 monoclonal antibodies. Mouse anti-HRP2 monoclonal antibody was found to produce high optical density readings between dilutions of 1:100 and 1:3,200 by ELISA with assay signal observed up to a 1:200,000 dilution. nHRP2 yield from HB3 culture by bead-based immunoassay revealed that both culture supernatant and iRBC lysate were practical sources of large quantities of this antigen, producing a total yield of 292.4 µg of nHRP2 from two pooled culture preparations. Assessment of nHRP2 recognition by RDTs revealed that Carestart Pf HRP2 and HRP2/pLDH RDTs detected purified nHRP2 when applied at concentrations between 20.6 and 2060 ng/mL, performing within a log-fold dilution of commercially-available recombinant HRP2. The band intensity observed for the nHRP2 dilutions was equivalent to that observed for P. falciparum culture strain dilutions of 3D7 and US06 F Nigeria XII between 12.5 and 1000 parasites/µL. Conclusions: Purified nHRP2 could be a valuable reagent for laboratory applications as well as assessment of new and existing RDTs prior to their use in clinical settings. These results establish that it is possible to extract microgram quantities of the native HRP2 antigen from HB3 culture and that this purified protein is well recognized by existing monoclonal antibody lines and RDTs.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Synthesis and Characterization of Superconductor Diodes

In the era of modern computing, quantum bits (or qubits) are extensively researched due to their ability to perform computations exponentially faster than classical computers. One promising method for synthesizing qubits on a macroscopic scale is through superconductor diodes (SDs). Unfortunately, SDs are currently limited by their dependence on extremely low temperatures (T ≈ 0K), which restricts their practical usability at larger scales. To address this limitation, researchers are exploring new materials and methods to identify superconductors with higher critical temperatures. My research at the Lawrence Livermore National Laboratory (LLNL) focuses on synthesizing and characterizing superconductor diodes to determine their efficiency for quantum computing applications. Bismuth thin films were synthesized with magnetron sputtering with separate thicknesses which provides information on methods to alter their resistivity. Bismuth was characterized as the topological insulator used in the diode, and its resistivity was measured using a standard four-point probe technique with 1 mA current. Bismuth (99.99% pure) was then purified in a tubular furnace at 400C to use within a Josephson junction as a topological insulator. Niobium and Tantalum Josephson junctions were then synthesized, and they were characterized using a lock-in amplifier four-point probe technique. Materials such as niobium and tantalum are used in high-entropy alloy research, which is a relatively new class of material that has little research within the superconductor diode community, yet the current research shows promise. We anticipate that testing high-entropy superconductors will reveal crucial factors, potentially leading to the development of high-temperature superconductors in the future.

36 MATERIALS SCIENCE↗
DOE OSTI2020

Microbiological, Physicochemical, and Immunological Analysis of a Commercial Cashew Nut-Based Yogurt

Nut-based milks and yogurts are gaining popularity, but may not offer the same benefits as dairy yogurts to consumers. Cashew nuts often cause severe allergic reactions, and cashew nut allergens are stable to several types of processing. To compare its characteristics to dairy yogurt and characterize the effects of fermentation on the Ana o 1–3 cashew nut allergens, a commercial yogurt made from cashew nuts (Cashewgurt) was evaluated for microbiological, physiochemical, and immunological properties. Average counts for lactobacilli and Streptococcus thermophilus were greater than 10 million colony forming units per milliliter, indicating the capacity to provide a health benefit. Cashewgurt pH and viscosity values were comparable to cow milk yogurts, and it was off white in color. SDS-PAGE analysis indicated a clear reduction in Ana o 1 and 2, and immuno-assay with polyclonal anti-cashew IgG antibody and cashew-allergic IgE indicated an overall reduction in allergen content. In contrast, SDS-PAGE, mass spectrometry, immunoblot, and ELISA all revealed that Ana o 3 was relatively unaffected by the fermentation process. In conclusion, Ana o 1 and Ana o 2 are sensitive to degradation, while Ana o 3 survives lactic acid bacterial fermentation during yogurt production. The analysis presented here indicates that cashew nut yogurt is not suitable for those with cashew nut allergy.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

A “Graft to” Electrospun Zwitterionic Bilayer Membrane for the Separation of Hydraulic Fracturing-Produced Water via Membrane Distillation

Simultaneous fouling and pore wetting of the membrane during membrane distillation (MD) is a major concern. In this work, an electrospun bilayer membrane for enhancing fouling and wetting resistance has been developed for treating hydraulic fracture-produced water (PW) by MD. These PWs can contain over 200,000 ppm total dissolved solids, organic compounds and surfactants. The membrane consists of an omniphobic surface that faces the permeate stream and a hydrophilic surface that faces the feed stream. The omniphobic surface was decorated by growing nanoparticles, followed by silanization to lower the surface energy. An epoxied zwitterionic polymer was grafted onto the membrane surface that faces the feed stream to form a tight antifouling hydration layer. The membrane was challenged with an aqueous NaCl solution containing sodium dodecyl sulfate (SDS), an ampholyte and crude oil. In the presence of SDS and crude oil, the membrane was stable and displayed salt rejection (>99.9%). Further, the decrease was much less than the base polyvinylidene difluoride (PVDF) electrospun membrane. The membranes were also challenged with actual PW. Our results highlight the importance of tuning the properties of the membrane surface that faces the feed and permeate streams in order to maximize membrane stability, flux and salt rejection.

omniphobic↗
DOE OSTI2020

Amphiphilic Block Copolymers for Flocculation and Hydrophobization of Legacy Waste Suspensions in Flotation Driven Dewatering Operations - 20162

Flotation has been effectively demonstrated as a low footprint, high efficiency separation process when utilising simple anionic surface modifying agents such as sodium dodecyl sulphate (SDS). These agents increase hydrophobicity to remove suspended cationic surface charged particulates such as Mg(OH){sub 2} from waste suspensions similar to those at British nuclear fuel management sites- where corrosion processes have degraded fuel cladding. The technology is advantageous as it could effectively be retrofitted into existing waste management facilities at nuclear waste management sites such Sellafield and Hanford as the simplicity (no moving parts) and size of this technology coupled with low cost of construction could be rapidly deployed to aid in risk and hazard reduction decommissioning operations with minimum impact to secondary waste generation. Previously, sodium dodecyl sulphate has been deployed as a collector agent to increase the hydrophobicity of Mg(OH){sub 2} suspended particles, achieving a maximum particulate extraction percentage of 93%. To achieve further particulate extraction, flotation performance has been determined to be a function of the hydrodynamic limitations extracting fine material (>20 μm). Fine particulates have been shown in the minerals industry to have an inability to overcome slipstreams created from rising bubbles in flotation cells, likely due to the low mass and thus momentum of the colloidal material, preventing interaction between the hydrophobized particles and air-water interfaces (bubbles). To combat this mass transfer resistance poly(acrylic acid)-block-poly(n-butyl acrylate) amphiphilic diblock copolymers were synthesised via reversible addition-fragmentation chain-transfer (RAFT) polymerisation. The anticipated mode of action of these polymers involves the hydrophilic poly(acrylic acid) block polymer acting as a flocculation agent to increase the mass of the aggregates, and the hydrophobic poly(n-butyl acrylate) modifies the hydrophobicity of the resulting flocs. The concentration of this polymer (termed a macro-collector) was varied and the change in particle size distribution of the particulate suspension was analysed using static light scattering. The flotation performance was then characterised by several methods including percentage particulate recovery, volume reduction factor and residual bulk particulate concentration. This was then benchmarked against the traditional surfactant collector agent SDS. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2021

Estimating reaction parameters in mechanism-enabled population balance models of nanoparticle size distributions: A Bayesian inverse problem approach

In order to quantitatively predict nano- as well as other particle-size distributions, one needs to have both a mathematical model and estimates of the parameters that appear in these models. Here, we show how one can use Bayesian inversion to obtain statistical estimates for the parameters that appear in recently derived mechanism-enabled population balance models (ME-PBM) of nanoparticle growth. The Bayesian approach addresses the question of “how well do we know our parameters, along with their uncertainties?.” The results reveal that Bayesian inversion statistical analysis on an example, prototype $\mathrm{lr(0)_n}$ nanoparticle formation system allows one to estimate not just the most likely rate constants and other parameter values, but also their SDs, confidence intervals, and other statistical information. Moreover, knowing the reliability of the mechanistic model's parameters in turn helps inform one about the reliability of the proposed mechanism, as well as the reliability of its predictions. Importantly, the paper can also be seen as a tutorial with the additional goal of achieving a “Gold Standard” Bayesian inversion ME-PBM benchmark that others can use as a control to check their own use of this methodology for other systems of interest throughout nature. Overall, the results provide strong support for the hypothesis that there is substantial value in using a Bayesian inversion methodology for parameter estimation in particle formation systems.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2022

High-yield recombinant bacterial expression of 13 C-, 15 N-labeled, serine-16 phosphorylated, murine amelogenin using a modified third generation genetic code expansion protocol

Amelogenin constitutes ~ 90% of the enamel matrix in the secretory stage of amelogenesis, a still poorly understood process that results in the formation of the hardest and most mineralized tissue in vertebrates – enamel. Most biophysical research with amelogenin uses recombinant protein expressed in Escherichia coli. In addition to providing copious amounts of protein, recombinant expression allows 13 C- and 15 N-labelling for detailed structural studies using NMR spectroscopy. However, native amelogenin is phosphorylated at one position, Ser-16 in murine amelogenin, and a lingering question overshadowing decades of enamel research with recombinant amelogenin is the effect of phosphorylation on its biophysical properties. Using a modified genetic code expansion protocol we have expressed and purified uniformly 13 C-, 15 N-labeled murine amelogenin (pS16M179) with ~95% of the protein being correctly phosphorylated. Homogeneous phosphorylation was achieved using commercially available, enriched, 13 C-, 15 N-labeled media and protein expression was induced with isopropyl β-D-1-thiogalactopyranoside at 310 K. Phosphoserine incorporation was verified from one-dimensional 31P NMR spectra, comparison of 1 H- 15 N HSQC spectra, Phos-tag SDS PAGE, and mass spectrometry. Phosphorus-31 NMR spectra for pS16M179 under conditions known to trigger amelogenin self-assembly into nanospheres confirm nanosphere models with buried N-termini. Lambda phosphatase treatment of these nanospheres results in the dephosphorylation of pS16M179, confirming that smaller oligomers and monomers with exposed N-termini are in equilibrium with nanospheres. Such 13 C-, 15 N-labeling of amelogenin with accurately encoded phosphoserine incorporation will accelerate biomineralization research to understand amelogenesis and stimulate the expanded use of genetic code expansion protocols to introduce phosphorylated amino acids into proteins.

31P-NMR↗
DOE OSTI2023

Comments on the entanglement spectrum of de Sitter space

We argue that the Schwarzschild-de Sitter black hole entropy formula does not imply that the entanglement spectrum of the vacuum density matrix of de Sitter space is flat. Specifically, we show that the expectation value of a random projection operator of dimension d >> 1, on a Hilbert space of dimension D >> d and in a density matrix ρ = e –K with strictly positive spectrum, is $\frac{d}{D} \large{(}1 + o \large{(}\frac{1}{\sqrt{d}}\large{)}$, independent of the spectrum of the density matrix. In addition, for a suitable class of spectra the asymptotic estimates Tr (ρK) ~ ln D – o(1) and Tr [ρ(K – $\langle$K$\rangle$) 2 ] = a $\langle$K$\rangle$ are compatible for any order one constant a. We discuss a simple family of matrix models and projections that can replicate such modular Hamiltonians and the SdS entropy formula.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2019

Cloning, Purification, and Characterization of Recombinant Thermostable β-Xylanase Tnap-0700 from Thermotoga naphthophila

The gene of a β-xylanase (Tnap-0700) was cloned from a hyperthermophilic Thermotoga naphthophila strain ATCC BAA-489 and expressed in Escherichia coli BL21 (DE3) via pET-21a (+) as an expression vector. The growth steps were upgraded for highest β-xylanase expression via several factors, for example, IPTG concentration, time of induction, pH, and temperature. The pH and temperature optima for the extreme expression of β-xylanase were 7.0 pH and 37 °C, correspondingly. Recombinant enzyme purified by heat treatment process, then later by immobilized metal ion affinity chromatography. Molecular mass of the purified β-xylanase was 38 kDa observed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was stable at room temperature for 30 days. It exhibited high stability over wide series of temperature 50–90 °C and pH 4.0–9.0 upon the addition of 1 mM Ca{sup +2} and reduced in the existence of Cu{sup +2} and EDTA. The addition of about 10–30% different organic solvents have no considerable effect on enzyme. However, SDSF and urea acting as an inhibitor leads to decrease in the enzyme activity. The β-xylanase enzyme was active to hydrolyze xylan from beechwood forming xylose. Thermostable β-xylanase causes the breakdown of complex carbohydrates into monosaccharide components. This thermostable β-xylanase revealed remarkable properties, which make it an encouraging candidate for various industrial applications especially in the alteration of renewable biomaterials into ethanol production, and biofuels from lignocellulosics has acknowledged much devotion subsequently in the last decade.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2021

Effects of anionic and nonionic surfactants on the dispersion and stability of nanoSiO2 in aqueous and cement pore solutions

It has been well recognized that the benefits and effectiveness of nanoparticles in cement-based materials could not be maximized if these are not well dispersed. To address this issue, in this study, different anionic (SDS and PCE) and nonionic surfactants (Tweens and Tritons) were used to disperse nanosilica (NS) in aqueous solution and cement pore solution. The results show that the dispersibility of NS in cement pore solution was improved, and the compressive strength of the cement-NS pastes increased linearly with critical micelle concentration (CMC) of nonionic surfactants. Among all surfactants studied, Triton X-405 led the paste to the highest increase in strength (33% at 1-day and 41% at 3-days) since it had the highest CMC. TEM and EDS analysis evidenced that this strength increase might be attributed to the nucleation of outer product CSH gel and its densification with calcite nanocrystals, attributed to Triton X-405 addition.

36 MATERIALS SCIENCE↗
DOE OSTI2023

Stability study of aqueous foams under high-temperature and high-pressure conditions relevant to Enhanced Geothermal Systems (EGS)

In Enhanced Geothermal System (EGS) hydraulic fracturing is carried out by injecting cold water into deep Hot Dry Rocks (HDR) under carefully controlled conditions to create new or reopen existing fractures. Water-based fracturing fluids demonstrate some challenges including immense quantity of water usage, water sensitivity of the formations, water blocking, and lack of proppant carrying capacity and transportation. Thus, an alternative is to use foam-based fracturing fluid which offers potential advantage over conventional water-based fracturing fluid such as minimum water usage, reduced wellbore damage, high proppant carrying capacity, and less environmental damage. However, foams are complex mixture of gaseous phase and liquid phase which are thermodynamically unstable at downhole conditions, and their stability can decrease over time due to liquid drainage, bubble coarsening, and coalescence. This paper shows laboratory experiments executed to study foam stability at high temperature (200°C) and high pressure (6.9MPa) conditions which simulates the geothermal environment. Foam stability was characterized by half-life of foam, which is defined as the time taken by the foam to decreases by 50% of its original height due to drainage. In this paper, two types of gaseous phases, nitrogen (N 2 ) and carbon dioxide (CO 2 ) were investigated. Also, based on successful practice of foam-based fracturing fluid in oil and gas industries, four surfactants, including Alpha olefin sulfonate (AOS), Sodium dodecyl sulfonate (SDS), Tergitol™ (NP – 40), and Cetyltrimethylammonium chloride (CTAC) at optimum concentration of 1wt.% were tested for best stability performance. In addition, different stabilizing agents including guar gum, bentonite clay, crosslinker, silicon dioxide nanoparticles (SiO 2 ), graphene oxide (GO) were also studied. Experimental results showed that N 2 foams were more stable than CO 2 foams. It was observed that foam half-life decreased with the increase in temperature. Among all the surfactants, AOS foams showed the most promising thermal stability at high temperatures. Moreover, with the addition of stabilizing agents, foam's half-life was enhanced. Stabilizing agents such as crosslinker and GO dispersion showed the most stable foams with half-life recorded at 20min and 17min, respectively, at 200°C and 6.9MPa. Finally, pressure also showed a positive effect on foam stability; with increased pressure, foam half-life was increased. Based on the experimental data, analytical models for the effect of temperature and pressure were developed, considering foam degradation is a first-order kinetic reaction that linearly depends on the foam drainage mechanism. The effect of temperature on foam half-life was studied as an exponential decay model. In this model, foam half-life is a function of drainage rate constant (D A ) and activation energy (E a ) of the foam system. The effect of pressure on foam half-life was found to obey a power-law model where an increase in pressure showed an increase in foam half-life. Furthermore, a linear relation was studied for the effect of pressure on foam activation energy and drainage rate. Then the combined effects of temperature and pressure were studied, which yielded an analytical model to predict the foam stabilities in terms of half-life for different foam compositions. In conclusion, this research indicates that with an appropriate selection of surfactants and stabilizing agents, it is possible to obtain stable foams, which could replace conventional water fracturing fluid under EGS conditions.

15 GEOTHERMAL ENERGY↗
DOE OSTI2025

Characterizing the pore-scale impacts of enhanced-solubilization solutions on organic-liquid morphology and fluid-fluid interfacial area with microtomography

Synchrotron X-ray microtomography was used to directly quantify the impact of enhanced-solubilization solutions on mobilization of a non-wetting fluid in a natural porous medium. Polyoxyethylene sorbitan monooleate (tween 80), hydroxypropyl-β-cyclodextrin (HPCD), sodium dodecyl sulfate (SDS), and ethanol were used as the solubilization reagents. Tetrachloroethene (PCE) was used as the entrapped organic immiscible liquid. Microtomography images were collected prior to and after successive floods with each reagent solution. The total trapping number was varied by more than three orders of magnitude by changing the velocity and viscosity of the aqueous phase as well as the interfacial tension between fluid phases. The results were compared to those obtained from equivalent experiments conducted with water flooding. Inspection of images indicated that mobilization caused a change in PCE morphology and distribution for all reagents with the exception of HPCD, for which a lower trapping number was determined compared to the other experiments. Mobilization of organic liquid was observed at total trapping numbers that were consistent with prior work. The enhanced-solubilization solutions had minimal impact on the configuration or magnitude of fluid-fluid interfacial area.

Interfacial area↗
DOE OSTI2021

Simplification of the purification of heat stable recombinant low molecular weight proteins and peptides from GST-fusion products

The synthesis and purification of peptides of importance in the fields of research and medicine continue to be a challenging task. Chemical synthesis of oligopeptides, especially those greater than 25 amino acids, is cost prohibitive. On the other hand, several bottlenecks exist in the production of recombinant short peptides in heterologous expression hosts such as Escherichia coli (E. coli). In this paper, a rapid, cost-effective, and reliable method for the production and singlestep-purification of peptides and small proteins was developed. Five peptides and small proteins were overexpressed in E. coli as GST-fusion products in high yields. The recombinant peptides or proteins were successfully purified after enzymatic cleavage with selective heat-induced precipitation of the GST-affinity tag. Qualitative and quantitative analysis using SDS-PAGE and mass spectrometric methods suggest that the recombinant peptides/ proteins were purified to >95% homogeneity. Results of biophysical experiments, including multi-dimensional NMR spectroscopy, show that the purified proteins/ peptides retain their native conformation. Isothermal titration calorimetry studies indicate no significant change in the binding affinity of the heattreated purified product to their interacting partner(s) compared to the recombinant peptides purified by conventional chromatographic procedures without subjecting to heat treatment. In our opinion, the results reported render the purification of recombinant proteins/ peptides of biomedical relevance using our proposed method easy and reliable.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Construction of extracellular matrix-based 3D hydrogel and its effects on cardiomyocytes

Highlights: • SOD and Triton X-100 could efficiently decellularize cells to get cardiac ECM. • Main proteins of cECM were fibronectin, collagen, elastin, fibrillin, laminin, etc. • 75% cECM-containing 3D hydrogel promoted cell proliferation and anti-apoptosis. • cECM3DH facilitated the differentiation of ESCs into CMs. • APcECM3DH was more conducive to NCM and ESC-derived CM maturation. Some discoveries resulted from 2-dimensional (2D) cultured cardiac cells have been disqualified in animal testing and later clinical trials. Extracellular matrix (ECM) plays a vital role in cardiac homeostasis, cardiac ECM (cECM)-based 3D cell cultures can mimics the physiological and pathological conditions in vivo closely, it is hopeful of addressing this challenge. Construction of cECM-based 3-dimensional (3D) hydrogel (cECM3DH) and its effects on cell behaviors were studied here. The results indicated that cellular compartments could be efficiently removed from heart tissue via sodium dodecyl sulfonate (SDS)- and Triton X-100-mediated decellularization, remaining the natural fibrous network structure and major proteins. 3D hydrogel consisted of 1 × 10{sup 7} cells/mL cells and 75% cECM could promote the proliferation and anti-apoptosis ability of human embryonic kidney (HEK)-293T cells. 0.25% trypsin or 0.20% collagenase was suitable to retrieve these cells from 3D hydrogel for further researches. Compared with 2D culture system, cECM3DH could significantly increase the proportion of GATA 4{sup +} cardiomyocytes (CMs) derived from heart tissue of neonatal mouse or induced differentiation of embryonic stem cells (ESCs) (P < 0.05) The expression levels of mature genes including cTnT, JCN, CaV1.2, MYL2, CASQ2, NCX1, and Cx43 of these CMs in adult pig cECM-based 3D hydrogel (APcECM3DH) were significantly higher than that in 2D culture system and in newborn piglet cECM-based 3D hydrogel (NPcECM3DH), respectively (P < 0.05). Therefore, cECM3DH supports the generation of primary CMs and ESC-derived CMs, APcECM3DH was more conducive to promoting CM maturation, which contributes to building 3D model for pathogenesis exploration, drug screening, and regenerative medicine of heart diseases.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2023

Antibodies Raised Against an Aβ Oligomer Mimic Recognize Pathological Features in Alzheimer’s Disease and Associated Amyloid-Disease Brain Tissue

Antibodies that target the β-amyloid peptide (Aβ) and its associated assemblies are important tools in Alzheimer’s disease research and have emerged as promising Alzheimer’s disease therapies. This paper reports the creation and characterization of a triangular Aβ trimer mimic composed of Aβ 17–36 β-hairpins and the generation and study of polyclonal antibodies raised against the Aβ trimer mimic. The Aβ trimer mimic is covalently stabilized by three disulfide bonds at the corners of the triangular trimer to create a homogeneous oligomer. Structural, biophysical, and cell-based studies demonstrate that the Aβ trimer mimic shares characteristics with oligomers of full-length Aβ. X-ray crystallography elucidates the structure of the trimer and reveals that four copies of the trimer assemble to form a dodecamer. SDS-PAGE, size exclusion chromatography, and dynamic light scattering reveal that the trimer also forms higher-order assemblies in solution. Cell-based toxicity assays show that the trimer elicits LDH release, decreases ATP levels, and activates caspase-3/7 mediated apoptosis. Immunostaining studies on brain slices from people who lived with Alzheimer’s disease and people who lived with Down syndrome reveal that the polyclonal antibodies raised against the Aβ trimer mimic recognize pathological features including different types of Aβ plaques and cerebral amyloid angiopathy.

60 APPLIED LIFE SCIENCES↗