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At least 73 records · Page 4

Fundamentals of bio-electrochemical sensing

The utilization of bio-functionalities such as biorecognition or catalysis derived them their name biosensors. Bio-electrochemical sensing is a new discipline that combines the advantages of biological detection and electrochemical transduction. Bio-electrochemical sensors are devices that use biological materials such as enzymes, antibodies, DNA, or cells as receptors to detect target analytes in a variety of samples. Electrodes convert biological interactions into electrical impulses, which can be studied using various electrochemical techniques. Bio-electrochemical sensors have demonstrated significant promise for use in clinical diagnostics, environmental monitoring, food safety, and biotechnology. Biosensors have received numerous applications in recent years because they are fast, simple, and inexpensive for practical applications. In this article, we cover the basic principles, design strategies, immobilization and regeneration techniques, along with the advantages and applications of bio-electrochemical sensors. Finally, this article discusses the rationale for developing electrochemical biosensors in the context of the various bio-receptors that can be applied.

36 MATERIALS SCIENCE↗

Isotope Exchange between Mercuric [Hg(II)] Chloride and Hg(II) Bound to Minerals and Thiolate Ligands: Implications for Enriched Isotope Tracer Studies

Enriched mercury (Hg) stable isotopes have been widely used as tracers in field and laboratory investigations of Hg(II) biogeochemical transformations such as methylation and demethylation. Few studies, however, have considered concurrent isotope exchange reactions between newly spiked and pre-existing Hg(II) in environmental matrices, which may alter redistribution and thus transformation of the spiked and pre-existing Hg(II). Using enriched 198 Hg [as mercuric Hg(II) or HgCl n species], this study investigated isotope exchange between 198 Hg and pre-existing Hg(II) bound to metacinnabar (β-HgS), sediments, low-molecular-weight (LMW) thiols, and dissolved organic matter (DOM). The impact of isotope exchange on methylmercury production in the presence of organic ligands was also evaluated with an iron-reducing bacterium Geobacter sulfurreducens PCA in a phosphate buffered solution (pH 7.4). Here, we found that spiked 198 Hg readily exchanged with mineral-bound ambient Hg(II) despite concurrent Hg(II) adsorption and immobilization on the solids. Rapid exchange (<3 min) was also observed between spiked 198 Hg and 200 Hg pre-equilibrated with LMW thiols and DOM in solution. While the exchange did not cause net changes in Hg(II) chemical speciation, it resulted in redistribution of Hg(II) isotopes bound to the ligands and thus an apparently similar methylation rate and magnitude of the spiked 198 Hg and pre-existing 200 Hg by PCA cells when 198 Hg and 200 Hg were added at 1:1 ratio. These observations underscore the importance of isotope exchange when an enriched Hg isotope is applied in environmental matrices, as the exchange could potentially lead to biased rate calculations of Hg(II) transformation and bioaccumulation and thus risk assessments of new Hg(II) input to the natural ecosystems.

58 GEOSCIENCES↗

Comparative transcriptomics provides insights into molecular mechanisms of zinc tolerance in the ectomycorrhizal fungus Suillus luteus

Zinc (Zn) is a major soil contaminant and high Zn levels can disrupt growth, survival, and reproduction of fungi. Some fungal species evolved Zn tolerance through cell processes mitigating Zn toxicity, although the genes and detailed mechanisms underlying mycorrhizal fungal Zn tolerance remain unexplored. To fill this gap in knowledge, we investigated the gene expression of Zn tolerance in the ectomycorrhizal fungus Suillus luteus. We found that Zn tolerance in this species is mainly a constitutive trait that can also be environmentally dependent. Zinc tolerance in S. luteus is associated with differences in the expression of genes involved in metal exclusion and immobilization, as well as recognition and mitigation of metal-induced oxidative stress. Differentially expressed genes were predicted to be involved in transmembrane transport, metal chelation, oxidoreductase activity, and signal transduction. Some of these genes were previously reported as candidates for S. luteus Zn tolerance, while others are reported here for the first time. Our results contribute to understanding the mechanisms of fungal metal tolerance and pave the way for further research on the role of fungal metal tolerance in mycorrhizal associations.

59 BASIC BIOLOGICAL SCIENCES↗

Understanding the structure and composition of recalcitrant oligosaccharides in hydrolysate using high-throughput biotin-based glycome profiling and mass spectrometry

Novel Immunological and Mass Spectrometry Methods for Comprehensive Analysis of Recalcitrant Oligosaccharides in AFEX Pretreated Corn Stover. Lignocellulosic biomass is a sustainable alternative to fossil fuel and is extensively used for developing bio-based technologies to produce products such as food, feed, fuel, and chemicals. The key to these technologies is to develop cost competitive processes to convert complex carbohydrates present in plant cell wall to simple sugars such as glucose, xylose, and arabinose. Since lignocellulosic biomass is highly recalcitrant, it must undergo a combination of thermochemical treatment such as Ammonia Fiber Expansion (AFEX), dilute acid (DA), Ionic Liquid (IL) and biological treatment such as enzyme hydrolysis and microbial fermentation to produce desired products. However, when using commercial fungal enzymes during hydrolysis, only 75–85% of the soluble sugars generated are monomeric sugars, while the remaining 15–25% are soluble recalcitrant oligosaccharides that cannot be easily utilized by microorganisms. Previously, we successfully separated and purified the soluble recalcitrant oligosaccharides using a combination of charcoal and celite-based separation followed by size exclusion chromatography and studies their inhibitory properties on enzymes. We discovered that the oligosaccharides with higher degree of polymerization (DP) containing methylated uronic acid substitutions were more recalcitrant towards commercial enzyme mixtures than lower DP and neutral oligosaccharides. Here, we report the use of several complementary techniques that include glycome profiling using plant biomass glycan specific monoclonal antibodies (mAbs) to characterize sugar linkages in plant cell walls and enzymatic hydrolysate, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) using structurally-informative diagnostic peaks offered by negative ion post-secondary decay spectra, gas chromatography followed by mass spectrometry (GC–MS) to characterize oligosaccharide sugar linkages with and without derivatization. Since oligosaccharides (DP 4–20) are small, it is challenging to mobilize these molecules for mAbs binding and characterization. To overcome this problem, we have applied a new biotin-coupling based oligosaccharide immobilization method that successfully tagged most of the low DP soluble oligosaccharides on to a micro-plate surface followed by specific linkage analysis using mAbs in a high-throughput system. This new approach will help develop more advanced versions of future high throughput glycome profiling methods that can be used to separate and characterize oligosaccharides present in biomarkers for diagnostic applications.

59 BASIC BIOLOGICAL SCIENCES↗

Dimolybdenum Paddlewheel Complexes with Cation Binding Sites as Electrolyte Additives to Manipulate the Solid-Electrolyte Interphase at Lithium Metal Anodes

The use of electrolyte additives at millimolar loadings to control the surface chemistry of lithium metal anodes (LMAs) is a leading strategy to improve lithium metal batteries and promote electrosynthetic reactions. Whereas previous studies employed either inorganic or organic additives, in this study, we report the first organometallic additive, Mo 2 (mea) 4 [1, mea = 2-(2-methoxyethoxy)acetate], a dimolybdenum paddlewheel complex that is stable under Li plating conditions and features cation binding sites in the second coordination sphere that promote reversible Li + coordination. Binding of Li + ions to 1 induces immobilization of cationically charged aggregates (or products thereof) into the solid electrolyte interphase (SEI), imparting multiple beneficial functions. The modified SEI was found to protect the LMA against parasitic side reactions, produce modest but measurable improvements to Li plating properties (e.g., overpotential, surface structure, and Coulombic efficiency), and improve interfacial charge transport properties. Furthermore, the most notable benefit to battery cycling performance appears in calendar aging tests, which show that the presence of the additive protects the LMA from parasitic side reactions that would otherwise decrease overall cell cycling efficiency over time. Collectively, these data disclose a tactic for designing electrolyte additives using principles of organometallic synthesis.

Additives↗

Molecularly tunable heterogeneous catalysts by edge functionalization of graphitic carbons

Disclosed are surface immobilized (electro)catalysts that may be prepared by a condensation reaction that generates an aromatic unit that is robust to acid and base and elevated temperatures. Among their many desirable characteristics, the catalysts are far less prone to the bimolecular deactivation pathways commonly observed for homogeneous catalysts, and may be used in solvents with a range of polarities and dielectric strengths. The catalysts are suitable for a wide array of thermal catalytic reactions (polymerization, oxidation, hydrogenation, cross-coupling etc.) and as anodes and/or cathodes in fuel cells, electrolyzers, and in batteries and supercapacitors.

Surendranath, Yogesh↗

Mechanically and Thermally Robust Gel Electrolytes Built from A Charged Double Helical Polymer

Polymer electrolytes have received tremendous interest in the development of solid-state batteries, but often fall short in one or more key properties required for practical applications. Herein, a rigid gel polymer electrolyte prepared by immobilizing a liquid mixture of a lithium salt and poly(ethylene glycol) dimethyl ether with only 8 wt% poly(2,2′-disulfonyl-4,4′-benzidine terephthalamide) (PBDT) is reported. The high charge density and rigid double helical structure of PBDT lead to formation of a nanofibrillar structure that endows this electrolyte with stronger mechanical properties, wider temperature window, and higher battery rate capability compared to all other poly(ethylene oxide) (PEO)-based electrolytes. The ion transport mechanism in this rigid polymer electrolyte is systematically studied using multiple complementary techniques. Li/LiFePO 4 cells show excellent capacity retention over long-term cycling, with thermal cycling reversibility between ambient temperature and elevated temperatures, demonstrating compelling potential for solid-state batteries targeting fast charging at high temperatures and slower discharging at ambient temperature.

36 MATERIALS SCIENCE↗

Thermal Process Technology for Nuclear Applications - 20379

ANSTO's Synroc technology has been developed to provide a safe, secure matrix for the immobilization and final disposal of radioactive waste. Synroc technology will be used to manage radioactive wastes from the production of the radioisotope Molybdenum-99 (Mo-99). This paper shall outline various stages of the process development with specific reference to the thermal treatment technology of calcination. Calcination is a key step in the Synroc process [1-2].The rotary thermal processing system includes: an advanced heating element design for increased robustness and ease of remote operation and maintenance, an enhanced modular design of components for ease of remote maintenance in a hot cell and in compliance with hot cell radioactive environment requirements for safety, reliability and maintainability. In addition to thermal treatment of waste from nuclear medicine production, this technology provides solutions for a variety of nuclear materials processing applications including sintering UO{sub 2} pellets for reactor fuel rods, oxidation of UO{sub 2} pellets, swarf, and powder to U{sub 3}O{sub 8}, de-nitration of Uranyl nitrate and hydrofluorination of UO{sub 2} pellets. The paper will also discuss thermal processing solutions for a range of nuclear applications. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Solutal-buoyancy-driven intertwining and rotation of patterned elastic sheets

Abstract The intertwining of strands into 3D spirals is ubiquitous in biology, enabling functions from information storage to maintenance of cell structure and directed locomotion. In synthetic systems, entwined fibers can provide superior mechanical properties and act as artificial muscle or structural reinforcements. Unlike structures in nature, the entwinement of synthetic materials typically requires application of an external stimulus, such as mechanical actuation, light, or a magnetic field. Herein, we use computational modeling to design microscale sheets that mimic biology by transducing chemical energy into mechanical action, and thereby self-organize and interlink into 3D spirals, which spontaneously rotate. These flexible sheets are immersed in a fluid-filled microchamber that encompasses an immobilized patch of catalysts on the bottom wall. The sheets themselves can be passive or active (coated with catalyst). Catalytic reactions in the solution generate products that occupy different volumes than the reactants. The resulting density variations exert a force on the fluid (solutal buoyancy force) that causes motion, which in turn drives the interlinking and collective swirling of the sheets. The individual sheets do not rotate; rotation only occurs when the sheets are interlinked. This level of autonomous, coordinated 3D structural organization, intertwining, and rotation is unexpected in synthetic materials systems operating without external controls. Using physical arguments, we identify dimensionless ratios that are useful in scaling these ideas to other systems. These findings are valuable for creating materials that act as “machines”, and directing soft matter to undergo self-sustained, multistep assembly that is governed by intrinsic chemical reactions.

Manna, Raj Kumar (ORCID:0000000322729487)↗

Journal Pre-proof3D printed polylactic acid and acrylonitrile butadiene styrene fluidicstructures for biological applications: tailoring bio-material interfaceviasurface modification

Three-dimensional (3D) printing is a rapidly growing technology that best fits for cell culture and other biological applications due to its potential to build immensely complex structures from customized designs. However, use of 3D printed structures for cell adhesion, proliferation, and activation requires tailoring of surface characteristics. In this context, this work investigates the use of two biocompatible 3D printable polymer materials, Poly-L-lactic Acid (PLA) and Acrylonitrile Butadiene Styrene (ABS) for bio applications, and the effect of their post-printing surface modification processes to achieve desired bio functionality. Three post-printing surface modification techniques, alkaline hydrolysis, ultraviolet ozone plasma irradiation and gold thin film deposition are performed with the objective of introducing useful levels of surface functionalities. Poly-L-lysine (PLL) labelled with FITC chromophore is immobilized on the surface modified samples following standard protocols. The effect of surface roughness and porosity of the 3D printed structures on the polypeptide immobilization is compared on as-printed versus mechanically polished surfaces. Different characterization methods, viz., Fluorescence microscopy, Raman Spectroscopy and UV–vis spectroscopy, are used to study the efficacy of the surface modification and PLL immobilization techniques. Our results demonstrated denser PLL attachment on polished hydrolyzed PLA and ABS surfaces which enables the use of these printable polymers for a range of applications such as tissue scaffolds and microfluidics.

3D Printing, PLA, ABS, Biodegradable Polymer, Surf↗

Accelerating Biomimetic Solar - Energy Harvesting: Mapping the Interaction Landscape of Plasmonic-Excitonic Hybrid Nanosystems (Final Report)

In general, excitonic and plasmonic nanoscale materials in close proximity show high potential for significant breakthroughs in energy related materials research. The interactions between these two kinds of materials result in coupled optical transitions (plexcitons), distinct from those of both the individual exciton and plasmon as well as from those of the sum of their constituents (synergistic effects). By linking together materials-research and physical-research approaches, this project contributes to a concerted approach on nanomaterials energy research. The project’s overall goal is to accelerate the development of well-defined plexcitonic model systems consisting of carefully engineered plasmonic and excitonic nanomaterial— essential for both gaining a fundamental understanding of plexcitonic nanomaterials and the development of novel design principles for biomimetic solar energy harvesting. During the 3-year project period and the terminal renewal with limited support for a 12-month period, we successfully synthesized and characterized (1) a robust excitonic nanomaterial and (2) a library of plasmonic nanoparticles as well as developed (3) a microfluidic platform for homogenous nanosynthesis as summarized below: (1) Robust Excitonic Nanomaterial. Supramolecular assemblies are Nature’s most successful material system for solar energy harvesting. However, photovoltaic devices based on artificial supramolecular assemblies continue to be stymied by disappointing efficiencies and poor stability. The conceptual failure may lie in current solar cell architectures, which rely on solidifying supramolecular assemblies as an ensemble into a solid matrix, neglecting the intrinsic fragility of the assemblies’ internal structure, thus disrupting or even destroying their delicate optoelectronic properties, that is, delicate Frenkel excitonic properties. Supramolecular assemblies may finally serve as usable light harvesting material systems for solar energy conversion technologies, only if they meet the following criteria: (a) Stability, that is, the fragile structure including its delicate Frenkel excitonic character needs to be stable, (b) Robustness, that is, resistant against elevated and fluctuating temperatures, and (c) Viability for device integration, that is, capable of being immobilized onto solid substrates. Here, by developing a nanocomposite via a tunable, cage-like scaffold design, we successfully provided stable supramolecular nanocomposites, that inhabit robust Frenkel excitons despite harming environmental conditions such as extreme heat stress. (2) Library of Plasmonic Nanoparticles. Naturally, current models describing plasmonic hybrid quantum states—plasmonic hybridizations—parallel those developed for molecular orbitals, equating individual plasmonic nanostructures with “atoms” and the plasmonic nanoassemblies with “molecules.” In analogy to organic synthesis, a suitably robust fabrication method would allow for “atom-like” manipulation of “molecule-like” plasmonic nanoassemblies; of high value for next-generation energy nanotechnologies. Despite this frequent comparison, current plasmonic nanoassembly fabrication methods favor top-down templating over wet-chemical synthesis, however, achieving precise control over nanostructure’s geometry and surface characteristics remain an art and a scientific challenge. The conceptual failure may lie in the current wet-chemical synthesis paradigm, as it relies on the accessibility of a multi-dimensional synthesis parameter space through limited, rather one-dimensional synthesis procedures by employing step-by-step approaches. Solution-based nanoarchitectonics for rational design of precisely built plasmonic nanoassemblies via solution-based fabrication may finally be possible only if multi-dimensional syntheses approaches are available that allow for comprehensive control over the plasmonic nanomaterials’ (a) Structural Properties and (b) Surface Properties. Here, by developing an innovative multidimensional 1,3-propanediol based polyol synthesis, we successfully provided control over the plasmonic building-block’s geometry (size and shape) together with its surface characteristics. Our results present a critical step toward the vision of a “periodic table-like” system for plasmonic materials based on straightforward wet-chemical syntheses for energy nanotechnologies. Developing deliberate modifications on this synthesis, we generated a library of plasmonic nanostructures covering the vast parameter space—opening the door for fundamental investigation of plexcitonic model systems. (3) Microfluidic Platform for Homogenous Nanosynthesis. Control over structural properties of plexcitonic nanocomposites remains a challenge due to current limitations in nanosynthesis techniques. Slight variations in nanostructure’s geometry impact their optoelectronic properties, demanding precise synthesis beyond the capabilities of solution-based (batch) synthesis processes. In contrast, the small, confined liquid volumes used in microfluidics—a reaction technique where the manipulation of fluids takes place in channels with dimensions of tens of micrometers—allows for homogenous synthesis conditions, providing excellent control of the reaction and, as a result, of the materials’ geopmetry and composition. However, thus far, the majority of plexcitonic systems has been developed via batch synthesis. Here, by successfully developing a two-channel microreactor, our microfluidic-supported synthesis approach combines the advantages of both microfluidics and batch platforms, allowing for precise spatio-temporal control over all synthesis parameters opening the possibility for homogenous nanosythnesis of well-defined plexcitonic model systems.

14 SOLAR ENERGY↗

Adsorption and intracellular uptake of mercuric mercury and methylmercury by methanotrophs and methylating bacteria

The cell surface adsorption and intracellular uptake of mercuric mercury Hg(II) and methylmercury (MeHg) are important in determining the fate and transformation of Hg in the environment. However, current information is limited about their interactions with two important groups of microorganisms, i.e., methanotrophs and Hg(II)-methylating bacteria, in aquatic systems. This study investigated the adsorption and uptake dynamics of Hg(II) and MeHg by three strains of methanotrophs, Methylomonas sp. strain EFPC3, Methylosinus trichosporium OB3b, and Methylococcus capsulatus Bath, and two Hg(II)-methylating bacteria, Pseudodesulfovibrio mercurii ND132 and Geobacter sulfurreducens PCA. Distinctive behaviors of these microorganisms towards Hg(II) and MeHg adsorption and intracellular uptake were observed. The methanotrophs took up 55–80% of inorganic Hg(II) inside cells after 24 h incubation, lower than methylating bacteria (>90%). Approximately 80–95% of MeHg was rapidly taken up by all the tested methanotrophs within 24 h. In contrast, after the same time, G. sulfurreducens PCA adsorbed 70% but took up <20% of MeHg, while P. mercurii ND132 adsorbed <20% but took up negligible amounts of MeHg. These results suggest that microbial surface adsorption and intracellular uptake of Hg(II) and MeHg depend on the specific types of microbes and appear to be related to microbial physiology that requires further detailed investigation. Despite being incapable of methylating Hg(II), methanotrophs play important roles in immobilizing both Hg(II) and MeHg, potentially influencing their bioavailability and trophic transfer. Furthermore, methanotrophs are not only important sinks for methane but also for Hg(II) and MeHg and can influence the global cycling of C and Hg.

59 BASIC BIOLOGICAL SCIENCES↗

Design of a Single-Ion Conducting Polymer Electrolyte for Sodium-Ion Batteries

Here, a sodium bis(fluoroallyl)malonato borate salt (NaBFMB) is synthesized. Using a Click thiol-ene reaction, NaBFMB can be photo-crosslinked with a tri-thiol (trimethylolpropane tris(3-mercapto propionate), TMPT) to create a single-ion conducting electrolyte (NaSIE), with all negative charges residing on the borate moieties and anions immobilized through the 3-D crosslinked network. The NaSIE can be prepared either as a free-standing film or through a drop-cast method followed by a photo crosslinking method for an in-situ formation on top of the electrodes. The free-standing film of NaSIE has a high ionic conductivity of 2 × 10 –3 S cm –1 at 30 °C, and a high transference number (t Na + ) of 0.91 as measured through the Bruce-Vincent method. The electrochemical stability of NaSIE polymer electrolyte is demonstrated via cyclic voltammetry (CV) to be stable up to 5 V vs Na/Na + . When tested inside a symmetrical Na//Na cell, the NaSIE shows a critical current density (CCD) of 0.4 mA cm –2 . The stability of NaSIE is further demonstrated via a long cycling of the stripping/plating test with a current density of 0.1 mA cm –2 at five-minute intervals for over 10,000 min. Using the in-situ method, NaSIE is used as the electrolyte for a sodium metal battery using P2 (Na resides at prismatic sites with with ABBAAB stacking)-cathode of Na 0.67 Ni 0.33 Mn 0.67 O 2 (NNMO) and is cycled between the cut-off voltages of 2.0–4.0 V. A high initial specific capacity (85.7 mAh g –1 ) with a capacity retention of 86.79% after 150 cycles is obtained.

25 ENERGY STORAGE↗

A Versatile Method for Creating Ultrathin Films of Polyzwitterions with Antifouling Properties

Protein repellent (i.e., antifouling) properties of ultrathin polymeric films have been ascribed to the formation of a dense layer of adsorbed polymers, which causes steric hindrance to protein adsorption on underlying substrates. The antifouling properties of the films are hypothesized to be independent of polymer hydrophilicity based on previous studies involving uncharged polymers. Here, to test the hypothesis and extend it to the realm of charged polymers, we have developed a grafting method for creating ultrathin layers of polyzwitterions on solid surfaces. To demonstrate the efficacy of this method, poly[1-(3-sulfopropyl-1)-2-vinylpyridinium betaine] (P2VPPS) was synthesized via free-radical polymerization and immobilized onto surfaces by using a photoreactive benzophenone derivative as the cross-linker, which was covalently bonded to either quartz or Si/SiOx substrates through a silane anchor. We found that ultrathin films with thicknesses of less than 10 nm can be obtained in dry states using 365 nm ultraviolet (UV) cured/cross-linked surface coatings. These films were found to be hydrophilic and undergo structural rearrangements in the presence of water. The antifouling performance of the films was evaluated using Pseudomonas aeruginosa strain PAO1 bacterium as the fouling agent and exhibited antifouling properties with a kinetic control via the coating method (i.e., spin-coating and drop-casting). In particular, the surface area coverages due to the PAO1 cell attachment on the quartz substrates coated with P2VPPS were found to be ∼5% and ∼0% for the spin-coated and drop-casted films, respectively, after 4 h. These results show that the grafting method can be used to generate surfaces with antifouling properties based on ultrathin films of polyzwitterions.

Pseudomonas aeruginosa↗

Assessment of the Corrosion Risks to a Double Shell Tank During the Selective Removal of a Cesium-rich Supernate Layer - 20227

The Direct Feed Low Activity Waste (DF-LAW) activity will provide the initial feed to the Waste Treatment and Immobilization Plant at the Hanford Tank Farm facility. Feed preparation for DF-LAW will be carried out in multiple double shell waste tanks (DST) utilizing the Tank Side Cesium Removal (TSCR) process. Tank AP-106 will be an integral part of the TSCR process and hence key to the DFLAW activity. Prior to being utilized for this purpose, the Cs-137 concentration levels in the AP-106 supernate must be reduced significantly. The AP-106 re-purposing process manages normal waste tank addition procedures such that waste acceptance criteria objectives for Cs-137 removal are achieved. As this process was being developed, several concerns regarding tank integrity during and after repurposing of Tank AP-106, and the associated tanks (i.e., AP-102, AN-101, and AP-108), were addressed. The potential corrosion degradation mechanisms identified based on process knowledge included: general and pitting corrosion, caustic stress corrosion cracking, concentration cell corrosion, and microbiologically induced corrosion (MIC). A systematic approach was undertaken to assess the risk of corrosion during the AP-106 re-purposing process. The assessment considered the initial and present condition of the tank, the service history of the tank, the environmental conditions created in each tank during each stage of the process, process timing, and corrosion testing and modeling that approximated tank conditions. Based on this information, recommendations were provided to minimize the exposure of the tank to corrosive conditions. These recommendations included time limits for each stage of the process, staging of corrosion inhibitor solutions should process delays occur, and waste sampling with potential chemistry adjustments at defined steps in the process. Additionally, recommendations for post-process supernate chemistry adjustments and tank inspections were put forth. The greatest risk for this process occurs during the manipulation of large quantities of Columbia River water (i.e., greater than 450,000 gallons) within AP-106, the associated tanks and the supporting infrastructure (i.e., transfer line piping). This risk becomes more pronounced if there is a process upset and the tank walls remain exposed to a corrosive condition longer than intended. As long as the river water is blended into the supernate in a timely fashion, and the waste is adjusted appropriately with corrosion inhibitors, corrosion may be minimized and the final condition of the supernate in AP-106 and the associated tanks is expected to be in compliance with corrosion chemistry requirements. Contingency plans will be made to minimize the impact of a process upset. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Self-Assembling Metabolon Enables the Cell Free Conversion of Glycerol to 1,3-Propanediol

Cell free biocatalysis is showing promise as a replacement or complement to conventional microbial biocatalysts due to the potential for achieving high yields, titers, and productivities. However, there exist several challenges that need to be addressed before its broader industrial adoption is achieved. New paradigms and innovative solutions are needed to overcome these challenges. In this study we demonstrate high levels of glycerol conversion to 1,3-propanediol using a self-assembling metabolic pathway leveraging the arraying strategy (protein scaffolds) used by thermophilic cellulolytic bacteria to assemble their biomass degrading enzymes. These synthetic metabolons were capable of producing 1,3-PDO at a yield more than 95% at lower glycerol concentration and close to 70% at higher concentrations at a higher productivity rate than the equivalent microbial strain. One of the benefits of our approach is the fact that no enzyme purification is required, and that the assembly of the complex is accomplished in vivo before immobilization, while product formation is conducted in vitro. We also report the recovery of enzymatic activity upon fusion enzymes binding to these protein scaffolds, which could have broader applications when assembling arrayed protein complexes.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Resolving SPARC–HSA binding kinetics with an ultrasensitive photonic sensor based on bound states in the continuum

Secreted protein acidic and rich in cysteine (SPARC) is critical in cell-matrix interactions and tissue remodeling. It influences tumor progression through its affinity for human serum albumin (HSA) - the most abundant plasma protein, which also plays a crucial role in drug delivery. Strong molecular binding leads to a dissociation constant KD in the nanomolar range. Thus, determining KD requires detecting sub-nanomolar concentrations with ultrasensitive methods. This may be crucial for elucidating the nature of SPARC-HSA binding, as their interaction remains a subject of debate. Capturing these interactions accurately requires a platform capable of resolving rapid binding kinetics at extremely low analyte concentrations. In this work, we report on a microfluidics-integrated photonic nanostructure that supports bound states in the continuum (BICs) and is optimized for studying the fast kinetics of high-affinity protein-protein interactions. The unprecedented capability of detecting sub-nanomolar concentrations allows quantifying KD between SPARC and HSA beyond the state of the art. We leverage an all-dielectric photonic crystal slab (PhCS) sustaining two BIC branches arising from gapped Dirac cone dispersion. HSA is covalently immobilized on the PhCS bonded to a PDMS microfluidic chamber. SPARC dissociation is carried out using PBS buffer (pH 7.4), ensuring complete protein release through precise control of the flow rate and continuous spectral monitoring of the BICs. The measured KD=8.2±0.8 nM confirms the strong affinity of SPARC for HSA. This study highlights the potential of BIC-based sensing as a versatile tool for investigating protein interactions. These results also have implications for the optimization of drug delivery systems and cancer treatment strategies.

Albumin↗

Analysis of a macrophage carbamylated proteome reveals a function in post-translational modification crosstalk

Background. Lysine carbamylation is a biomarker of rheumatoid arthritis and kidney diseases. However, its cellular function is understudied due to the lack of tools for systematic analysis of this post-translational modification (PTM). Methods. We adapted a method to analyze carbamylated peptides by co-affinity purification with acetylated peptides based on the cross-reactivity of anti-acetyllysine antibodies. We also performed immobilized-metal affinity chromatography to enrich for phosphopeptides, which allowed us to obtain multi-PTM information from the same samples. Results. By testing the pipeline with RAW 264.7 macrophages treated with bacterial lipopolysaccharide, 7,299, 8,923 and 47,637 acetylated, carbamylated, and phosphorylated peptides were identified, respectively. Our analysis showed that carbamylation occurs on proteins from a variety of functions on sites with similar as well as distinct motifs compared to acetylation. To investigate possible PTM crosstalk, we integrated the carbamylation data with acetylation and phosphorylation data, leading to the identification 1,183 proteins that were modified by all 3 PTMs. Among these proteins, 54 had all 3 PTMs regulated by lipopolysaccharide and were enriched in immune signaling pathways, and in particular, the ubiquitin-proteasome pathway. We found that carbamylation of linear diubiquitin blocks the activity of the anti-inflammatory deubiquitinase OTULIN. Conclusions. Overall, our data show that anti-acetyllysine antibodies can be used for effective enrichment of carbamylated peptides. Moreover, carbamylation may play a role in PTM crosstalk with acetylation and phosphorylation, and that it is involved in regulating ubiquitination in vitro.

59 BASIC BIOLOGICAL SCIENCES↗