Far-Field Approximation in the Generalized Geometry Holdup (GGH) Model

Quantitative gamma spectrometry measurements of uranium frequently require corrections for attenuation by an equipment or container layer and by the uranium bearing material itself. It is common to correct for attenuation using the ''far-field approximation''. Under this approximation, the minimum thickness of equipment or material is used for the correction rather than an average thickness over the detector field-of-view. In reality this aspect of the far-field approximation is really a narrow field-of-view approximation. The price of this simplification is the introduction of a bias. This bias will be investigated in this paper. In addition, there is a distance dependence of the radial response of a detector. This dependence will also be investigated

The Self Attenuation Correction for Holdup Measurements, a Historical Perspective

Self attenuation has historically caused both conceptual as well as measurement problems. The purpose of this paper is to eliminate some of the historical confusion by reviewing the mathematical basis and by comparing several methods of correcting for self attenuation focusing on transmission as a central concept

Filter Paper: Solution to High Self-Attenuation Corrections in HEPA Filter Measurements

An 8 by 8 by 6 inch High Efficiency Particulate Air (HEPA) filter was measured as part of a uranium holdup survey in June of 2005 as it has been routinely measured every two months since 1998. Although the survey relies on gross gamma count measurements, this was one of a few measurements that had been converted to a quantitative measurement in 1998. The measurement was analyzed using the traditional Generalized Geometry Holdup (GGH) approach, using HMS3 software, with an area calibration and self-attenuation corrected with an empirical correction factor of 1.06. A result of 172 grams of {sup 235}U was reported. The actual quantity of {sup 235}U in the filter was approximately 1700g. Because of this unusually large discrepancy, the measurement of HEPA filters will be discussed. Various techniques for measuring HEPA filters will be described using the measurement of a 24 by 24 by 12 inch HEPA filter as an example. A new method to correct for self attenuation will be proposed for this measurement Following the discussion of the 24 by 24 by 12 inch HEPA filter, the measurement of the 8 by 8 by 6 inch will be discussed in detail

Comparison of the NDA of HEU Oxide between the AWCC and the HPGe Detector

This paper compares the performance of the Active Well Coincidence Counter (AWCC) with the performance of high resolution gamma spectrometry using an HPGe detector to nondestructively assay highly enriched (HEU) oxide. Traditionally the AWCC was considered to be the more appropriate instrument for this measurement. Although the AWCC had a high degree of precision, the HPGe provided the more accurate measurement of this material. The AWCC determines mass of U-235 from the coincident pairs of neutron detections, or doubles rate. The HPGe determines the mass of both U-235 and U238, the enrichment, and the quantity of other radioisotopes. The Tl-208 gamma rays were used to verify the amount of attenuation for the HPGe analysis. Fifty-four cans of enriched U3O8 were shipped to the Y-12 National Security Complex from Los Alamos National Laboratory (LANL) under Scrap Declaration LANL-45. The declared values for net weight, mass of uranium, mass of U-235, and enrichment (percent mass of U-235 to total uranium) are shown in Table A-1. The masses of U-235 range from 104g to 2404g and the enrichment varies from 20% to 98%

Rational Design and Characterization of D-Phe-Pro-D-Arg-Derived Direct Thrombin Inhibitors

The tremendous social and economic impact of thrombotic disorders, together with the considerable risks associated to the currently available therapies, prompt for the development of more efficient and safer anticoagulants. Novel peptide-based thrombin inhibitors were identified using in silico structure-based design and further validated in vitro. The best candidate compounds contained both l- and d-amino acids, with the general sequence d-Phe(P3)-Pro(P2)-d-Arg(P1)-P1′-CONH2. The P1′ position was scanned with l- and d-isomers of natural or unnatural amino acids, covering the major chemical classes. The most potent non-covalent and proteolysis-resistant inhibitors contain small hydrophobic or polar amino acids (Gly, Ala, Ser, Cys, Thr) at the P1′ position. The lead tetrapeptide, d-Phe-Pro-d-Arg-d-Thr-CONH2, competitively inhibits α-thrombin's cleavage of the S2238 chromogenic substrate with a Ki of 0.92 µM. In order to understand the molecular details of their inhibitory action, the three-dimensional structure of three peptides (with P1′ l-isoleucine (fPrI), l-cysteine (fPrC) or d-threonine (fPrt)) in complex with human α-thrombin were determined by X-ray crystallography. All the inhibitors bind in a substrate-like orientation to the active site of the enzyme. The contacts established between the d-Arg residue in position P1 and thrombin are similar to those observed for the l-isomer in other substrates and inhibitors. However, fPrC and fPrt disrupt the active site His57-Ser195 hydrogen bond, while the combination of a P1 d-Arg and a bulkier P1′ residue in fPrI induce an unfavorable geometry for the nucleophilic attack of the scissile bond by the catalytic serine. The experimental models explain the observed relative potency of the inhibitors, as well as their stability to proteolysis. Moreover, the newly identified direct thrombin inhibitors provide a novel pharmacophore platform for developing antithrombotic agents by exploring the conformational constrains imposed by the d-stereochemistry of the residues at positions P1 and P1′

Development of a versatile laboratory experiment to teach the metabolic transformation of hydrolysis

In this paper we describe an easy, reliable, versatile and inexpensive laboratory experiment to teach the metabolic transformation of hydrolysis to Pharmacy students. The experiment does not require the sacrifice of any experimental animal, or any work with organs or tissues, and so can be implemented in a typical university chemistry laboratory. We used acetylsalicylic acid (ASA), hexyl salicylate (HS) and two enzymes, a lipase and an esterase. Since both ASS and HS liberate salicylic acid (SA) upon hydrolysis, students can evaluate the different enzymatic transformations by monitoring the amount of SA liberated. The learning outcomes are an enhanced student understanding of: (1) the process of hydrolysis; (2) the application of enzymatic transformations of molecules from food to xenobiotics; (3) the differences between the general specificity of substrate of both enzymes; (4) the concepts of the lipophilic pocket; (5) the catalytic triad and its regioselectivity in relation to the ester bond. A questionnaire was administered to participating students at three points in time: at the beginning of the module, after enzymatic hydrolysis was taught in class, and after the laboratory experiment. From an analysis of the questionnaire data we conclude that this practical helped Pharmacy students to understand these concepts

Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli

Triglycerides (TAGs), the major storage molecules of metabolic energy and source of fatty acids, are produced as single cell oil by some oleogenic microorganisms. However, these microorganisms require strict culture conditions, show low carbon source flexibilities, lack efficient genetic modification tools and in some cases pose safety concerns. TAGs have essential applications such as behaving as a source for added-value fatty acids or giving rise to the production of biodiesel. Hence, new alternative methods are urgently required for obtaining these oils. In this work we describe TAG accumulation in the industrially appropriate microorganism Escherichia coli expressing the heterologous enzyme tDGAT, a wax ester synthase/triacylglycerol:acylCoA acyltranferase (WS/DGAT). With this purpose, we introduce a codon-optimized gene from the thermophilic actinomycete Thermomonospora curvata coding for a WS/DGAT into different E. coli strains, describe the metabolic effects associated to the expression of this protein and evaluate neutral lipid accumulation. We observe a direct relation between the expression of this WS/DGAT and TAG production within a wide range of culture conditions. More than 30% TAGs were detected within the bacterial neutral lipids in 90 minutes after induction. TAGs were observed to be associated with the hydrophobic enzyme while forming round intracytoplasmic bodies, which could represent a bottleneck for lipid accumulation in E. coli. We detected an increase of almost 3- fold in the monounsaturated fatty acids (MUFA) occurring in the recombinant strains. These MUFA were predominant in the accumulated TAGs achieving 46% of the TAG fatty acids. These results set the basis for further research on the achievement of a suitable method towards the sustainable production of these neutral lipids

Structure and catalytic regulatory function of ubiquitin specific protease 11 N-terminal and ubiquitin-like domains

The ubiquitin specific protease 11 (USP11) is implicated in DNA repair, viral RNA replication, and TGFβ signaling. We report the first characterization of the USP11 domain architecture and its role in regulating the enzymatic activity. USP11 consists of an N-terminal "domain present in USPs" (DUSP) and "ubiquitin-like" (UBL) domain, together referred to as DU domains, and the catalytic domain harboring a second UBL domain. Crystal structures of the DU domains show a tandem arrangement with a shortened β-hairpin at the two-domain interface and altered surface characteristics compared to the homologues USP4 and USP15. A conserved VEVY motif is a signature feature at the two-domain interface that shapes a potential protein interaction site. Small angle X-ray scattering and gel filtration experiments are consistent with the USP11DU domains and full-length USP11 being monomeric. Unexpectedly, we reveal, through kinetic assays of a series of deletion mutants, that the catalytic activity of USP11 is not regulated through intramolecular autoinhibition or activation by the N-terminal DU or UBL domains. Moreover, ubiquitin chain cleavage assays with all eight linkages reveal a preference for Lys(63)-, Lys(6)-, Lys(33)-, and Lys(11)-linked chains over Lys(27)-, Lys(29)-, and Lys(48)-linked and linear chains consistent with USP11's function in DNA repair pathways that is mediated by the protease domain. Our data support a model whereby USP11 domains outside the catalytic core domain serve as protein interaction or trafficking modules rather than a direct regulatory function of the proteolytic activity. This highlights the diversity of USPs in substrate recognition and regulation of ubiquitin deconjugation