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At least 235 records · Page 13

Capillary absorption spectroscopy for high temporal resolution measurements of stable carbon isotopes in soil and plant-based systems

Capillary Absorption Spectroscopy (CAS) is a relatively new analytical technique for performing stable isotope analysis. Here, we demonstrate the utility of CAS by recording and quantifying variation in 13C in controlled and biologically relevant applications. We calibrated CAS system response to increased 13CO2, with an observed ~4‰ increase in measured Δ13C for each 0.03 ppm shift in 13CO2 concentration. We leveraged this calibration to quantify rates of biogeochemical processes using a 13C tracer. For example, we monitored microbial respiration of 13C-glucose within an agricultural soil at 10 s quantification intervals and results demonstrated 8.6% ± 0.4 of added glucose was converted to 13CO2 within 1.5 h of incubation. We expanded the demonstration by adapting a rhizobox to permit continuous monitoring of 13CO2 in a soil (as distinct from plant) headspace to track the timing and quantify respiration rates of fresh plant photosynthate and observed a 3.5 h delay between plant exposure to a13CO2 tracer and the first signs of respiration by soil biota. These experiments highlight CAS is effective in producing high temporal resolution quantification of 13CO2 and demonstrate potential applications.

13CO2↗

Source Attribution of Cyanides using Anionic Impurity Profiling, Stable Isotope Ratios, Trace Elemental Analysis and Chemometrics

Chemical attribution signatures (CAS) for chemical threat agents (CTAs) are being investigated to provide an evidentiary link between CTAs and specific sources to support criminal investigations and prosecutions. In a previous study, anionic impurity profiles developed using high performance ion chromatography (HPIC) were demonstrated as CAS for matching samples from eight potassium cyanide (KCN) stocks to their reported countries of origin. Herein, a larger number of solid KCN stocks (n = 13) and, for the first time, solid sodium cyanide (NaCN) stocks (n = 15) were examined to determine what additional sourcing information can be obtained through anion, carbon stable isotope, and elemental analyses of cyanide stocks by HPIC, isotope ratio mass spectrometry (IRMS), and inductively coupled plasma optical emission spectroscopy (ICP-OES), respectively. The HPIC anion data was evaluated using the variable selection methods of Fisher-ratio (F-ratio), interval partial least squares (iPLS), and genetic algorithm-based partial least squares (GAPLS) and the classification methods of partial least squares discriminate analysis (PLSDA), K nearest neighbors (KNN), and support vector machines discriminate analysis (SVMDA). In summary, hierarchical cluster analysis (HCA) of anion impurity profiles from multiple cyanide stocks from six reported country of origins resulted in cyanide samples clustering into three groups: Czech Republic, Germany, and United States, independent of the associated alkali metal (K or Na). The three country groups were independently corroborated by HCA of cyanide elemental profiles and corresponded to countries with known solid cyanide factories. Both the anion and elemental CAS are believed to originate from the aqueous alkali hydroxides used in cyanide manufacture. Carbon stable isotope measurements resulted in two clusters: Germany and United States (the single Czech stock grouped with United States stocks). The carbon isotope CAS is believed to originate from the carbon source and process used to make the HCN utilized in cyanide synthesis. Classification errors for two validation studies using anion impurity profiles collected over five years on different instruments were as low as zero for KNN and SVMDA, demonstrating the excellent reliability (so far) of using anion impurities for matching a cyanide sample to its country of manufacture (i.e., factory). Variable selection reduced errors for those classification methods having errors greater than zero with iPLS-forward selection, and F-ratio typically providing the lowest errors. Finally, using anion profiles to match cyanides to a specific stock or stock group resulted in cross-validation errors ranging from zero to 5.3%.

chemical forensics↗

High energy resolution five-crystal spectrometer for high quality fluorescence and absorption measurements on an x-ray absorption spectroscopy beamline

Fluorescence detection is classically achieved with a solid state detector (SSD) on x-ray absorption spectroscopy (XAS) beamlines. This kind of detection however presents some limitations related to the limited energy resolution and saturation. Crystal analyzer spectrometers (CAS) based on a Johann-type geometry have been developed to overcome these limitations. We have tested and installed such a system on the BM30B/CRG-FAME XAS beamline at the ESRF dedicated to the structural investigation of very dilute systems in environmental, material and biological sciences. The spectrometer has been designed to be a mobile device for easy integration in multi-purpose hard x-ray synchrotron beamlines or even with a laboratory x-ray source. The CAS allows to collect x-ray photons from a large solid angle with five spherically bent crystals. It will cover a large energy range allowing to probe fluorescence lines characteristic of all the elements from Ca (Z = 20) to U (Z = 92). It provides an energy resolution of 1-2 eV. XAS spectroscopy is the main application of this device even if other spectroscopic techniques (RIXS, XES, XRS, etc.) can be also achieved with it. The performances of the CAS are illustrated by two experiments that are difficult or impossible to perform with SSD and the complementarity of the CAS vs SSD detectors is discussed.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Downfolding of many-body Hamiltonians using active-space models: Extension of the sub-system embedding sub-algebras approach to unitary coupled cluster formalisms

Here, we discuss the extension of the recently introduced subsystem embedding subalgebra coupled cluster (SES-CC) formalism to unitary CC formalisms. In analogy to the standard single-reference SES-CC formalism, its unitary CC extension allows one to include the dynamical (outside the active space) correlation effects in an SES induced complete active space (CAS) effective Hamiltonian. In contrast to the standard single-reference SES-CC theory, the unitary CC approach results in a Hermitian form of the effective Hamiltonian. Additionally, for the double unitary CC (DUCC) formalism, the corresponding CAS eigenvalue problem provides a rigorous separation of external cluster amplitudes that describe dynamical correlation effects—used to define the effective Hamiltonian—from those corresponding to the internal (inside the active space) excitations that define the components of eigenvectors associated with the energy of the entire system. The proposed formalism can be viewed as an efficient way of downfolding many-electron Hamiltonian to the low-energy model represented by a particular choice of CAS. In principle, this technique can be extended to any type of CAS representing an arbitrary energy window of a quantum system. The Hermitian character of low-dimensional effective Hamiltonians makes them an ideal target for several types of full configuration interaction type eigensolvers. As an example, we also discuss the algebraic form of the perturbative expansions of the effective DUCC Hamiltonians corresponding to composite unitary CC theories and discuss possible algorithms for hybrid classical and quantum computing. Given growing interest in quantum computing, we provide energies for H 2 and Be systems obtained with the quantum phase estimator algorithm available in the Quantum Development Kit for the approximate DUCC Hamiltonians.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Identifying candidate structured RNAs in CRISPR operons

Noncoding RNAs with secondary structures play important roles in CRISPR-Cas systems. Many of these structures likely remain undiscovered. We used a large-scale comparative genomics approach to predict 156 novel candidate structured RNAs from 36,111 CRISPR-Cas systems. A number of these were found to overlap with coding genes, including palindromic candidates that overlapped with a variety of Cas genes in type I and III systems. Among these 156 candidates, we identified 46 new models of CRISPR direct repeats and 1 tracrRNA. This tracrRNA model occasionally overlapped with predicted cas9 coding regions, emphasizing the importance of expanding our search windows for novel structure RNAs in coding regions. We also demonstrated that the antirepeat sequence in this tracrRNA model can be used to accurately assign thousands of predicted CRISPR arrays to type II-C systems. This study highlights the importance of unbiased identification of candidate structured RNAs across CRISPR-Cas systems.

59 BASIC BIOLOGICAL SCIENCES↗

A DETAILED KINEMATIC MAP OF CASSIOPEIA A'S OPTICAL MAIN SHELL AND OUTER HIGH-VELOCITY EJECTA

We present three-dimensional (3D) kinematic reconstructions of optically emitting material in the young Galactic supernova remnant Cassiopeia A (Cas A). These Doppler maps have the highest spectral and spatial resolutions of any previous survey of Cas A and represent the most complete catalog of its optically emitting material to date. We confirm that the bulk of Cas A's optically bright ejecta populate a torus-like geometry tilted approximately 30 Degree-Sign with respect to the plane of the sky with a -4000 to +6000 km s{sup -1} radial velocity asymmetry. Near-tangent viewing angle effects and an inhomogeneous surrounding circumstellar material/interstellar medium environment suggest that this geometry and velocity asymmetry may not be faithfully representative of the remnant's true 3D structure or the kinematic properties of the original explosion. The majority of the optical ejecta are arranged in several well-defined and nearly circular ring-like structures with diameters between approximately 30'' (0.5 pc) and 2' (2 pc). These ejecta rings appear to be a common phenomenon of young core-collapse remnants and may be associated with post-explosion input of energy from plumes of radioactive {sup 56}Ni-rich ejecta that rise, expand, and compress non-radioactive material. Our optical survey encompasses Cas A's faint outlying ejecta knots and exceptionally high-velocity NE and SW streams of S-rich debris often referred to as ''jets''. These outer knots, which exhibit a chemical make-up suggestive of an origin deep within the progenitor star, appear to be arranged in opposing and wide-angle outflows with opening half-angles of Almost-Equal-To 40 Degree-Sign.

79 ASTRONOMY AND ASTROPHYSICS↗

A LIGHT CURVE ANALYSIS OF CLASSICAL NOVAE: FREE-FREE EMISSION VERSUS PHOTOSPHERIC EMISSION

We analyzed light curves of seven relatively slower novae, PW Vul, V705 Cas, GQ Mus, RR Pic, V5558 Sgr, HR Del, and V723 Cas, based on an optically thick wind theory of nova outbursts. For fast novae, free-free emission dominates the spectrum in optical bands rather than photospheric emission, and nova optical light curves follow the universal decline law. Faster novae blow stronger winds with larger mass-loss rates. Because the brightness of free-free emission depends directly on the wind mass-loss rate, faster novae show brighter optical maxima. In slower novae, however, we must take into account photospheric emission because of their lower wind mass-loss rates. We calculated three model light curves of free-free emission, photospheric emission, and their sum for various white dwarf (WD) masses with various chemical compositions of their envelopes and fitted reasonably with observational data of optical, near-IR (NIR), and UV bands. From light curve fittings of the seven novae, we estimated their absolute magnitudes, distances, and WD masses. In PW Vul and V705 Cas, free-free emission still dominates the spectrum in the optical and NIR bands. In the very slow novae, RR Pic, V5558 Sgr, HR Del, and V723 Cas, photospheric emission dominates the spectrum rather than free-free emission, which makes a deviation from the universal decline law. We have confirmed that the absolute brightnesses of our model light curves are consistent with the distance moduli of four classical novae with known distances (GK Per, V603 Aql, RR Pic, and DQ Her). We also discussed the reason why the very slow novae are about ∼1 mag brighter than the proposed maximum magnitude versus rate of decline relation.

79 ASTRONOMY AND ASTROPHYSICS↗

HIGH-ENERGY PROPERTIES OF THE ENIGMATIC Be STAR γ CASSIOPEIAE

We present the results of a broadband X-ray study of the enigmatic Be star Gamma Cassiopeiae (herein γ Cas) based on observations made with both the Suzaku and INTEGRAL observatories. γ Cas has long been recognized as the prototypical example of a small subclass of Be stars with moderately strong X-ray emission dominated by a hot thermal component in the 0.5-12 keV energy range (L {sub x} ≈ 10{sup 32}-10{sup 33} erg s{sup –1}). This places them at the high end of the known luminosity distribution for stellar emission, but several orders of magnitude below typical accretion-powered Be X-ray binaries. The INTEGRAL observations spanned an eight-year baseline and represent the deepest measurement to date at energies above ∼50 keV. We find that the INTEGRAL data are consistent within statistics to a constant intensity source above 20 keV, with emission extending up to ∼100 keV, and that searches for all of the previously reported periodicities of the system at lower energies led to null results. We further find that our combined Suzaku and INTEGRAL spectrum, which we suggest is the most accurate broadband X-ray measurement of γ Cas to date, is fitted extremely well with a thermal plasma emission model with a single absorption component. We found no compelling need for an additional non-thermal high-energy component. We discuss these results in the context of a currently favored models for γ Cas and its analogs.

79 ASTRONOMY AND ASTROPHYSICS↗

THE ORIGIN OF THE PUZZLING HARD X-RAY EMISSION OF γ CASSIOPEIAE

Massive B and Be stars produce X-rays from shocks in high-velocity winds with temperatures of a few million degrees and maximum X-ray luminosities of ≈10{sup 31} erg s{sup −1}. Surprisingly, a sub-group of early Be stars exhibits ≥20 times hotter X-ray temperatures and ≥10 times higher X-ray luminosities than normal. This group of Be stars, dubbed γ-Cas analogs, contains about 10 known objects. The origin of this bizarre behavior has been extensively debated in the past decades. Two mechanisms have been put forward: accretion of circumstellar disk matter onto an orbiting white dwarf, or magnetic field interaction between the star and the circumstellar disk. We show here that the X-ray and optical emissions of the prototype of the class, γ-Cas, are very well correlated on year timescales with no significant time delay. Since the expected migration time from internal disk regions that emit most of the optical flux to the orbit of the companion star is several years, the simultaneity of the high energy and optical flux variations indicates that X-ray emission arises from close to the star. The systematic lack of magnetic field detection reported in recent spectro-polarimetric surveys of Be stars is consistent with the absence of strong magnetic wind braking in these fast spinning stars but place strong constraints on the possible origin of the magnetic field. We propose that in γ-Cas, the magnetic field emerges from equatorially condensed subsurface convecting layers, the thickness of which steeply increases with rotation rate, and that γ-Cas and its analogs are the most massive and closest to critical rotation Be stars.

79 ASTRONOMY AND ASTROPHYSICS↗

The novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity

To overcome CRISPR-Cas defense systems, many phages and mobile genetic elements (MGEs) encode CRISPR-Cas inhibitors called anti-CRISPRs (Acrs). Nearly all characterized Acrs directly bind Cas proteins to inactivate CRISPR immunity. Here, using functional metagenomic selection, we describe AcrIIA22, an unconventional Acr found in hypervariable genomic regions of clostridial bacteria and their prophages from human gut microbiomes. AcrIIA22 does not bind strongly to SpyCas9 but nonetheless potently inhibits its activity against plasmids. To gain insight into its mechanism, we obtained an X-ray crystal structure of AcrIIA22, which revealed homology to PC4-like nucleic acid–binding proteins. Based on mutational analyses and functional assays, we deduced that acrIIA22 encodes a DNA nickase that relieves torsional stress in supercoiled plasmids. This may render them less susceptible to SpyCas9, which uses free energy from negative supercoils to form stable R-loops. Modifying DNA topology may provide an additional route to CRISPR-Cas resistance in phages and MGEs.

59 BASIC BIOLOGICAL SCIENCES↗

Phage-Induced Expression of CRISPR-Associated Proteins is Revealed by Shotgun Proteomics in Streptococcus thermophilus

The CRISPR/Cas system, comprised of clustered regularly interspaced short palindromic repeats along with their associated (Cas) proteins, protects bacteria and archaea from viral predation and invading nucleic acids. While the mechanism of action for this acquired immunity is currently under investigation, the response of Cas protein expression to phage infection has yet to be elucidated. In this study, we employed shotgun proteomics to measure the global proteome expression in a model system for studying the CRISPR/Cas response: infection of S. thermophilus DGCC7710 with phage 2972. Host and viral proteins were simultaneously measured following inoculation at two different multiplicities of infection and across various time points using two-dimensional liquid chromatography tandem mass spectroscopy. Thirty-seven out of forty predicted viral proteins were detected, including all proteins of the structural virome and viral effector proteins. In total, 1,013 of 2,079 predicted S. thermophilus proteins were detected, facilitating the monitoring of host protein synthesis changes in response to virus infection. Importantly, Cas proteins from all four CRISPR loci in the S. thermophilus DGCC7710 genome were detected, including loci previously thought to be inactive. Many Cas proteins were found to be constitutively expressed, but several demonstrated increased abundance during peak infection, including the Cas9 proteins from the CRISPR1 and CRISPR3 loci, which are key players in the interference phase of the CRISPR/Cas response. Altogether, these results provide novel insights into the proteomic response of S. thermophilus, specifically CRISPR-associated proteins, upon phage 2972 infection.

59 BASIC BIOLOGICAL SCIENCES↗

Closure Report for Corrective Action Unit 566: EMAD Compound, Nevada National Security Site, Nevada with ROTCs 1-2, Revision 0

This Closure Report (CR) presents information supporting the closure of Corrective Action Unit (CAU) 566: EMAD Compound, Nevada National Security Site, Nevada. Corrective Action Unit 566 comprises Corrective Action Site (CAS) 25-99-20, EMAD Compound, located within Area 25 of the Nevada National Security Site. The purpose of this CR is to provide documentation supporting the completed corrective actions and provide data confirming that the closure objectives for CAU 566 were met. To achieve this, the following actions were performed: • Review the current site conditions, including the concentration and extent of contamination. • Implement any corrective actions necessary to protect human health and the environment. • Properly dispose of corrective action and investigation wastes. • Document Notice of Completion and closure of CAU 566 issued by the Nevada Division of Environmental Protection. From October 2010 through May 2011, closure activities were performed as set forth in the Streamlined Approach for Environmental Restoration Plan for CAU 566: EMAD Compound, Nevada National Security Site, Nevada. The purposes of the activities as defined during the data quality objectives process were as follows: • Determine whether contaminants of concern (COCs) are present. • If COCs are present, determine their nature and extent, implement appropriate corrective actions, and properly dispose of wastes. Analytes detected during the closure activities were evaluated against final action levels (FALs) to determine COCs for CAU 566. Assessment of the data from collected soil samples, and from radiological and visual surveys of the site, indicates the FALs were exceeded for polychlorinated biphenyls (PCBs), semivolatile organic compounds (SVOCs), and radioactivity. Corrective actions were implemented to remove the following: • Radiologically contaminated soil assumed greater than FAL at two locations • Radiologically contaminated soil assumed greater than FAL with lead shot • PCB-contaminated soil • Radiologically contaminated filters and equipment • Fuels, lubricants, engine coolants, and oils • Lead debris • Electrical and lighting components assumed to be potential source materials, including - fluorescent light bulbs - mercury switches (thermostats) - circuit boards - PCB-containing ballasts Closure of CAU 566 was achieved through a combination of removal activities and closure in place. Corrective actions to remove COCs, and known and assumed potential source materials, were implemented as was practical. The PCBs remaining at the site are bounded laterally, but not vertically, within CAS 25-99-20 based upon step-out sampling; the sources (e.g., PCB transformer oils, diesel fuel from locomotive reservoirs) have been removed; the practice of the application of PCB-containing oils for soil stabilization has ceased; and the COCs are not readily mobile in the environment. Closure in place is necessary, and future land use of the site will be restricted from intrusive activities. This will effectively eliminate inadvertent contact by humans with the contaminated media. The DOE, National Nuclear Security Administration Nevada Site Office, provides the following recommendations: • No further corrective action is required at CAS 25-99-20. • Closure in place of CAS 25-99-20. • A use restriction is required at CAU 566. • A Notice of Completion to the DOE, National Nuclear Security Administration Nevada Site Office, is requested from the Nevada Division of Environmental Protection for closure of CAU 566. • Corrective Action Unit 566 should be moved from Appendix III to Appendix IV of the Federal Facility Agreement and Consent Order.

54 ENVIRONMENTAL SCIENCES↗

Closure Report for Corrective Action Unit 116: Area 25 Test Cell C Facility, Nevada National Security Site, Nevada

This Closure Report (CR) presents information supporting closure of Corrective Action Unit (CAU) 116, Area 25 Test Cell C Facility. This CR complies with the requirements of the Federal Facility Agreement and Consent Order (FFACO) that was agreed to by the State of Nevada; the U.S. Department of Energy (DOE), Environmental Management; the U.S. Department of Defense; and DOE, Legacy Management (FFACO, 1996 [as amended March 2010]). CAU 116 consists of the following two Corrective Action Sites (CASs), located in Area 25 of the Nevada National Security Site: (1) CAS 25-23-20, Nuclear Furnace Piping and (2) CAS 25-41-05, Test Cell C Facility. CAS 25-41-05 consisted of Building 3210 and the attached concrete shield wall. CAS 25-23-20 consisted of the nuclear furnace piping and tanks. Closure activities began in January 2007 and were completed in August 2011. Activities were conducted according to Revision 1 of the Streamlined Approach for Environmental Restoration Plan for CAU 116 (U.S. Department of Energy, National Nuclear Security Administration Nevada Site Office [NNSA/NSO], 2008). This CR provides documentation supporting the completed corrective actions and provides data confirming that closure objectives for CAU 116 were met. Site characterization data and process knowledge indicated that surface areas were radiologically contaminated above release limits and that regulated and/or hazardous wastes were present in the facility.

54 ENVIRONMENTAL SCIENCES↗

Corrective Action Investigation Plan for Corrective Action Unit 366: Area 11 Plutonium Valley Dispersion Sites, Nevada National Security Site, Nevada, Revision 0

Corrective Action Unit 366 comprises the six corrective action sites (CASs) listed below: (1) 11-08-01, Contaminated Waste Dump No.1; (2) 11-08-02, Contaminated Waste Dump No.2; (3) 11-23-01, Radioactively Contaminated Area A; (4) 11-23-02, Radioactively Contaminated Area B; (5) 11-23-03, Radioactively Contaminated Area C; and (6) 11-23-04, Radioactively Contaminated Area D. These sites are being investigated because existing information on the nature and extent of potential contamination is insufficient to evaluate and recommend corrective action alternatives (CAAs). Additional information will be obtained by conducting a corrective action investigation before evaluating CAAs and selecting the appropriate corrective action for each CAS. The results of the field investigation will support a defensible evaluation of CAAs that will be presented in the Corrective Action Decision Document. The sites will be investigated based on the data quality objectives (DQOs) developed July 6, 2011, by representatives of the Nevada Division of Environmental Protection and the U.S. Department of Energy (DOE), National Nuclear Security Administration Nevada Site Office. The DQO process was used to identify and define the type, amount, and quality of data needed to develop and evaluate appropriate corrective actions for CAU 366. The presence and nature of contamination at CAU 366 will be evaluated based on information collected from a field investigation. Radiological contamination will be evaluated based on a comparison of the total effective dose (TED) at sample locations to the dose-based final action level (FAL). The TED will be calculated by summing the estimates of internal and external dose. Results from the analysis of soil samples collected from sample plots will be used to calculate internal radiological dose. Thermoluminescent dosimeters placed at each sample location will be used to measure external radiological dose. Based on historical documentation of the releases associated with the nuclear tests, it was determined that CASs 11-23-02, 11-23-03, and 11-23-04 will be investigated as one release site. The three test areas associated with these CASs are in close proximity; the devices tested were all composed of plutonium and enriched uranium; and the ground zeroes are all posted high contamination areas (HCAs). Because the device tested at CAS 11-23-01 was composed primarily of enriched uranium and the ground zero is not a posted HCA, the CAS will be investigated as a separate release. The DQO process also resulted in an assumption that TED within the HCAs and contaminated waste dumps exceeds the FAL and requires corrective action. A field investigation will be performed to define where TED exceeds the FAL and to determine whether other contaminants of concern are present at the site associated with other activities that took place at the site or from spills or waste discovered during the investigation. The presence and nature of contamination from other types of releases (such as migration and any potential releases discovered during the investigation) will be evaluated using soil samples collected from the locations most likely containing contamination, if present. Appendix A provides a detailed discussion of the DQO methodology and the DQOs specific to each CAS.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Closure Report for Corrective Action Unit 573: Alpha Contaminated Sites Nevada National Security Site, Nevada with ROTC 1

This Closure Report (CR) presents information supporting the closure of Corrective Action Unit (CAU) 573: Alpha Contaminated Sites, Nevada National Security Site, Nevada. CAU 573 comprises the two corrective action sites (CASs): 05-23-02-GMX Alpha Contaminated Are-Closure in Place and 05-45-01-Atmospheric Test Site - Hamilton- Clean Closure. The purpose of this CR is to provide justification and documentation supporting the recommendation that no further corrective action is needed for CAU 573 based on the implementation of the corrective actions. Corrective action activities were performed at Hamilton from May 25 through June 30, 2016; and at GMX from May 25 to October 27, 2016, as set forth in the Corrective Action Decision Document (CADD)/Corrective Action Plan (CAP) for Corrective Action Unit 573: Alpha Contaminated Sites; and in accordance with the Soils Activity Quality Assurance Plan, which establishes requirements, technical planning, and general quality practices. Verification sample results were evaluated against data quality objective criteria developed by stakeholders that included representatives from the Nevada Division of Environmental Protection and the DOE, National Nuclear Security Administration Nevada Field Office (NNSA/NFO) during the corrective action alternative (CAA) meeting held on November 24, 2015. Radiological doses exceeding the final action level were assumed to be present within the high contamination areas associated with CAS 05-23-02, thus requiring corrective action. It was also assumed that radionuclides were present at levels that require corrective action within the soil/debris pile associated with CAS 05-45-01. During the CAU 573 CAA meeting, the CAA of closure in place with a use restriction (UR) was selected by the stakeholders as the preferred corrective action of the high contamination areas at CAS 05-23-02 (GMX), which contain high levels of removable contamination; and the CAA of clean closure was selected by the stakeholders as preferred corrective action for the debris pile at CAS 05-45-01 (Hamilton). The closure in place was accomplished by posting signs containing a warning label on the existing contamination area fence line; and recording the FFACO UR and administrative UR in the FFACO database, the NNSA/NFO CAU/CAS files, and the management and operating contractor Geographic Information Systems. The clean closure was accomplished by excavating the soil/debris pile, disposing of the contents at the Area 5 Radioactive Waste Management Complex, and collecting verification samples. The corrective actions were implemented as stipulated in the CADD/CAP, and verification sample results confirm that the criteria for the completion of corrective actions have been met. Based on the implementation of these corrective actions, NNSA/NFO provides the following recommendations: No further corrective actions are necessary for CAU 573; The Nevada Division of Environmental Protection should issue a Notice of Completion to NNSA/NFO for closure of CAU 573; CAU 573 should be moved from Appendix III to Appendix IV of the FFACO.

54 ENVIRONMENTAL SCIENCES↗

2015 Annual Summary Report for the Area 3 and Area 5 RWMs

This annual summary report presents results and conclusions of the Fiscal Year (FY) 2015 review to assess the adequacy of the performance assessments (PAs) and composite analyses (CAs) in support of the Disposal Authorization Statements for the Area 3 and Area 5 Radioactive Waste Management Sites at the Nevada National Security Site (NNSS). The report presents results and conclusions of the FY 2015 review and determines the adequacy of the PAs and CAs. Operational factors (e.g., waste forms and containers, facility design, and waste receipts), closure plans, monitoring results, and research and development (R&D) activities were reviewed to determine the adequacy of the PAs. Likewise, the environmental restoration activities at the NNSS relevant to the sources of residual radioactive material that are considered in the CAs, the land-use planning, and the results of the environmental monitoring and R&D activities were reviewed to determine the adequacy of the CAs.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Closure Report for Corrective Action Unit 116: Area 25 Test Cell C Facility, Nevada National Security Site, Nevada with ROTC-1

CR loaded to this OSTI record. Just adding new file which includes CR plus new ROTC 1 and update the metadata to the following: This Closure Report (CR) presents information supporting closure of Corrective Action Unit (CAU) 116, Area 25 Test Cell C Facility. This CR complies with the requirements of the Federal Facility Agreement and Consent Order (FFACO) that was agreed to by the State of Nevada; the U.S. Department of Energy (DOE), Environmental Management; the U.S. Department of Defense; and DOE, Legacy Management (FFACO, 1996 [as amended March 2010]). CAU 116 consists of the following two Corrective Action Sites (CASs), located in Area 25 of the Nevada National Security Site: (1) CAS 25-23-20, Nuclear Furnace Piping and (2) CAS 25-41-05, Test Cell C Facility. CAS 25-41-05 consisted of Building 3210 and the attached concrete shield wall. CAS 25-23-20 consisted of the nuclear furnace piping and tanks. Closure activities began in January 2007 and were completed in August 2011. Activities were conducted according to Revision 1 of the Streamlined Approach for Environmental Restoration Plan for CAU 116 (U.S. Department of Energy, National Nuclear Security Administration Nevada Site Office [NNSA/NSO], 2008). This CR provides documentation supporting the completed corrective actions and provides data confirming that closure objectives for CAU 116 were met. Site characterization data and process knowledge indicated that surface areas were radiologically contaminated above release limits and that regulated and/or hazardous wastes were present in the facility. The Record of Technical Change 1 updated the use restriction information.

54 ENVIRONMENTAL SCIENCES↗

A Rapid Method for Detecting Normal or Modified Plant and Algal Carbonic Anhydrase Activity Using Saccharomyces cerevisiae

In recent years, researchers have attempted to improve photosynthesis by introducing components from cyanobacterial and algal CO2-concentrating mechanisms (CCMs) into terrestrial C3 plants. For these attempts to succeed, we need to understand the CCM components in more detail, especially carbonic anhydrase (CA) and bicarbonate (HCO3−) transporters. Heterologous complementation systems capable of detecting carbonic anhydrase activity (i.e., catalysis of the pH-dependent interconversion between CO2 and HCO3−) or active HCO3− transport can be of great value in the process of introducing CCM components into terrestrial C3 plants. In this study, we generated a Saccharomyces cerevisiae CA knock-out (ΔNCE103 or ΔCA) that has a high-CO2-dependent phenotype (5% (v/v) CO2 in air). CAs produce HCO3− for anaplerotic pathways in S. cerevisiae; therefore, the unavailability of HCO3− for neutral lipid biosynthesis is a limitation for the growth of ΔCA in ambient levels of CO2 (0.04% (v/v) CO2 in air). ΔCA can be complemented for growth at ambient levels of CO2 by expressing a CA from human red blood cells. ΔCA was also successfully complemented for growth at ambient levels of CO2 through the expression of CAs from Chlamydomonas reinhardtii and Arabidopsis thaliana. The ΔCA strain is also useful for investigating the activity of modified CAs, allowing for quick screening of modified CAs before putting them into the plants. CA activity in the complemented ΔCA strains can be probed using the Wilbur–Anderson assay and by isotope exchange membrane-inlet mass spectrometry (MIMS). Other potential uses for this new ΔCA-based screening system are also discussed.

59 BASIC BIOLOGICAL SCIENCES↗