Sample records for waste assay monitor

  1. Applications Where Snap is BPM for Radioactive Waste Assay

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Miller, T.J.

    2008-07-01

    Historically, the Atomic Weapons Establishment (AWE) at Aldermaston in the United Kingdom (UK), has used a variety of assay techniques to measure the radioactive content of a diverse range of waste packages from decommissioning, operational and legacy sources. The regulator, the Environment Agency in the UK, places conditions and limits on AWE through an authorisation within the Radioactive Substances Act (RSA93). The conditions and limits require Best Practical Means (BPM) measurements to be used to demonstrate compliance with the authorisation. Hence, the assay technique employed needs to achieve a balance between risk of exposure, environmental considerations, technological considerations, health andmore » safety considerations and cost effectiveness, without being grossly disproportionate in terms of money, time or trouble. Recently published work has concluded that the Spectral Non-destructive Assay Platform (SNAP) assay system is BPM for Depleted Uranium (DU) waste assay at AWE (1) and low level plutonium in soft drummed waste, HEPA filters and soils (2-4). The purpose of this paper is to highlight other applications where SNAP represents BPM for radioactive waste assay. This has been done by intercomparison studies of SNAP with other assay techniques, such as Segmented Gamma Scanner (SGS) and Passive Neutron Coincidence Counter (PNCC). It has been concluded that, for a large range of waste packages encountered at AWE, SNAP is BPM. (author)« less

  2. Assessment of the Microscreen phage-induction assay for screening hazardous wastes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Houk, V.S.; DeMarini, D.M.

    1987-09-01

    The Microscreen phage-induction assay, which quantitatively measures the induction of prophage lambda in Escherichia coli WP2s(lambda), was used to test 14 crude (unfractionated) hazardous industrial waste samples for genotoxic activity in the presence and absence of metabolic activation. Eleven of the 14 wastes induced prophage, and induction was observed at concentrations as low as 0.4 picograms per ml. Comparisons between the mutagenicity of these waste samples in Salmonella and their ability to induce prophage lambda indicate that the Microscreen phage-induction assay detected genotoxic activity in all but one of the wastes that were mutagenic in Salmonella. Moreover, the Microscreen assaymore » detected as genotoxic 5 additional wastes that were not detected in the Salmonella assay. The applicability of the Microscreen phage-induction assay for screening hazardous wastes for genotoxic activity is discussed along with some of the problems associated with screening highly toxic wastes containing toxic volatile compounds.« less

  3. Assessment of the microscreen phage-induction assay for screening hazardous wastes (1989)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Houk, V.S.; DeMarini, D.M.

    1989-01-01

    The Microscreen phage-induction assay, which quantitatively measures the induction of prophage Lambda in Escherichia coli WP2s(Lambda), was used to test 14 crude (unfractionated) hazardous industrial-waste samples for genotoxic activity in the presence and absence of metabolic activation. Eleven of the 14 wastes induced prophage, and induction was observed at concentrations as low as 0.4 picograms per ml. Comparisons of the mutagenic activity of these waste samples in Salmonella and their ability to induce prophage Lambda indicate that the phage-induction assay was a more-sensitive indicator of genetic damage for this group of wastes. All but one of the wastes that weremore » mutagenic to Salmonella were detected by the phage-induction assay, and 5 wastes not mutagenic to Salmonella were genetically active in the phage assay. The enhanced ability of the phage-induction assay to detect genotoxic activity may be related to the constituents comprising these waste samples. Partial chemical characterizations of the wastes showed high concentrations of carcinogenic metals, solvents, and chlorinated compounds, most of which are detected poorly by the Salmonella assay.« less

  4. Status of the waste assay for nonradioactive disposal (WAND) project

    NASA Astrophysics Data System (ADS)

    Arnone, Gaetano L.; Foster, Lynn A.; Foxx, Charles L.; Hagan, Roland C.; Martin, E. R.; Myers, Steven C.; Parker, Jack L.

    1999-01-01

    The WAND (Waste Assay for Nonradioactive Disposal) system scans thought-to-be-clean, low-density waste (mostly paper and plastics) to verify the absence of radioactive contaminants at very low-levels. Much of the low-density waste generated in radiologically controlled areas, formally considered `suspect' radioactive, is now disposed more cheaply at the Los Alamos County Landfill as opposed to the LANL Radioactive Waste Landfill.

  5. Pathology consultation on anticoagulation monitoring: factor X-related assays.

    PubMed

    Wool, Geoffrey D; Lu, Chuanyi M

    2013-11-01

    To review various anticoagulation therapies and related laboratory monitoring issues, with a focus on factor X-related chromogenic assays. A case-based approach is used to review pertinent published literatures and product inserts of anticoagulation drugs and to look back on clinical use of factor X-related chromogenic assays. The number of anticoagulants available to clinicians has increased greatly in the past decade. Whether and how these anticoagulants should be monitored are areas of uncertainty for clinicians, which can lead to misuse of laboratory assays and suboptimal patient management. Factor X-related assays are of particular concern because of the similar and often confusing test names. Based on a common clinical case scenario and literature review regarding anticoagulant monitoring, an up-to-date discussion and review of the various factor X-related assays are provided, focusing on the differences in test designs and clinical utilities between the chromogenic anti-Xa and chromogenic factor X activity assays. Anticoagulation therapy and related laboratory monitoring are rapidly evolving areas of clinical practices. A good knowledge of relevant laboratory assays and their clinical applications is necessary to help optimize patient care.

  6. Use of the microscreen phage-induction assay to assess the genotoxicity of 14 hazardous industrial wastes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Houk, V.S.; DeMarini, D.M.

    1988-01-01

    The Microscreen phage-induction assay, which quantitatively measures the induction of prophage lambda in Escherichia coli WP2s(lambda), was used to test 14 crude (unfractionated) hazardous industrial waste samples for genotoxic activity in the presence and absence of metabolic activation. Eleven of the 14 wastes induced prophage, and induction was observed at concentrations as low as 0.4 pg per ml. Comparisons between the ability of these waste samples to induce prophage and their mutagenicity in the Salmonella reverse mutation assay indicate that the phage-induction assay detected genotoxic activity in all but one of the wastes that were mutagenic in Salmonella. Moreover, themore » Microscreen assay detected as genotoxic five additional wastes that were not detected in the Salmonella assay. The applicability of the Microscreen phage-induction assay for screening hazardous wastes for genotoxic activity is discussed, as are some of the problems associated with screening highly toxic wastes containing toxic volatile compounds.« less

  7. Biotin-streptavidin enzyme-linked immunosorbent assay for detecting Tetrabromobisphenol A in electronic waste.

    PubMed

    Bu, Dan; Zhuang, Huisheng; Zhou, Xinchu; Yang, Guangxin

    2014-03-01

    Tetrabromobisphenol A (TBBPA) is a widely used brominated flame retardant. A sensitive and selective indirect competitive biotin-streptavidin-amplified enzyme-linked immunosorbent assay (BA-ELISA) was developed for detecting TBBPA. The optimal hapten of TBBPA was 2-(2,6-dibromo-4-(2-(3,5-dibromo-4-hydroxyphenly)propan-2-yl)) acetic acid. Several physiochemical factors that influence assay performance, such as optimal coupling concentration of immunogen and antibody, organic solvent, ionic strength, and pH, were studied and optimized. The limit of detection (IC10) was 0.027 ng/mL and the median inhibitory concentration (IC50) was 0.58 ng/mL. The BA-ELISA was highly selective, with low cross-reactivity with TBBPA analogs. Finally, the assay was used to detect TBBPA in electronic waste samples. The results are consistent with those using liquid chromatography, which proves that the proposed immunoassay is accurate and receptive. This BA-ELISA method is suitable for the rapid and sensitive screening of TBBPA in environmental monitoring. © 2013 Published by Elsevier B.V.

  8. Use of the Microscreen phage-induction assay to assess the genotoxicity of 14 hazardous industrial wastes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Houk, V.S.; DeMarini, D.M.

    1988-01-01

    The Microscreen phage-induction assay, which quantitatively measures the induction of prophage lambda in Escherichia coli WP2s lambda, was used to test 14 crude (unfractionated) hazardous industrial-waste samples for genotoxic activity in the presence and absence of metabolic activation. Eleven of the 14 wastes induced prophage, and induction was observed at concentrations as low as 0.4 picograms per ml. Comparisons between the mutagenicity of these waste samples in Salmonella and their ability to induce prophage lambda indicate that the Microscreen phage-induction assay detected genotoxic activity in all but one of the wastes that were mutagenic in Salmonella. Moreover, the Microscreen assaymore » detected as genotoxic 5 additional wastes that were not detected in the Salmonella assay. The applicability of the Microscreen phage-induction assay for screening hazardous wastes for genotoxic activity is discussed along with some of the problems associated with screening highly toxic wastes containing toxic volatile compounds.« less

  9. A batch assay to measure microbial hydrogen sulfide production from sulfur-containing solid wastes.

    PubMed

    Sun, Mei; Sun, Wenjie; Barlaz, Morton A

    2016-05-01

    Large volumes of sulfur-containing wastes enter municipal solid waste landfills each year. Under the anaerobic conditions that prevail in landfills, oxidized forms of sulfur, primarily sulfate, are converted to sulfide. Hydrogen sulfide (H2S) is corrosive to landfill gas collection and treatment systems, and its presence in landfill gas often necessitates the installation of expensive removal systems. For landfill operators to understand the cost of managing sulfur-containing wastes, an estimate of the H2S production potential is needed. The objective of this study was to develop and demonstrate a biochemical sulfide potential (BSP) test to measure the amount of H2S produced by different types of sulfur-containing wastes in a relatively fast (30days) and inexpensive (125mL serum bottles) batch assay. This study confirmed the toxic effect of H2S on both sulfate reduction and methane production in batch systems, and demonstrated that removing accumulated H2S by base adsorption was effective for mitigating inhibition. H2S production potentials of coal combustion fly ash, flue gas desulfurization residual, municipal solid waste combustion ash, and construction and demolition waste were determined in BSP assays. After 30days of incubation, most of the sulfate in the wastes was converted to gaseous or aqueous phase sulfide, with BSPs ranging from 0.8 to 58.8mLH2S/g waste, depending on the chemical composition of the samples. Selected samples contained solid phase sulfide which contributed to the measured H2S yield. A 60day incubation in selected samples resulted in 39-86% additional sulfide production. H2S production measured in BSP assays was compared with that measured in simulated landfill reactors and that calculated from chemical analyses. H2S production in BSP assays and in reactors was lower than the stoichiometric values calculated from chemical composition for all wastes tested, demonstrating the importance of assays to estimate the microbial sulfide production

  10. Monitoring of subsurface injection of wastes, Florida

    USGS Publications Warehouse

    Vecchioli, John

    1979-01-01

    Injection of waste liquids into Florida's subsurface is physically feasible in many places but should be accompanied by monitoring of the waste-receiving aquifer system in addition to the injection facility. Monitoring of the interaction of factors including hydrogeologic conditions, well construction, waste volumes and characteristics, and potable-water sources is desirable to assure that fresh-water resources are not being adversely affected. An effective aquifer-system monitoring program includes on-site wells located close to an injection well and open to the next-higher permeable stratum, satellite wells located hundreds to several thousands of feet from an injection well and open to the receiving aquifer, and regional wells located miles from individual injection wells and open to the receiving aquifer. An extensive aquifer-system monitoring program associated with two waste-injection facilities near Pensacola, Florida, has provided data which have aided hydrologists to understand the aquifer system's response to the injection and, accordingly, to evaluate the potential for affecting the area's fresh-water resources.

  11. A continuous spectrophotometric assay for monitoring adenosine 5'-monophosphate production.

    PubMed

    First, Eric A

    2015-08-15

    A number of biologically important enzymes release adenosine 5'-monophosphate (AMP) as a product, including aminoacyl-tRNA synthetases, cyclic AMP (cAMP) phosphodiesterases, ubiquitin and ubiquitin-like ligases, DNA ligases, coenzyme A (CoA) ligases, polyA deadenylases, and ribonucleases. In contrast to the abundance of assays available for monitoring the conversion of adenosine 5'-triphosphate (ATP) to ADP, there are relatively few assays for monitoring the conversion of ATP (or cAMP) to AMP. In this article, we describe a homogeneous assay that continuously monitors the production of AMP. Specifically, we have coupled the conversion of AMP to inosine 5'-monophosphate (IMP) (by AMP deaminase) to the oxidation of IMP (by IMP dehydrogenase). This results in the reduction of oxidized nicotine adenine dinucleotide (NAD(+)) to reduced nicotine adenine dinucleotide (NADH), allowing AMP formation to be monitored by the change in the absorbance at 340 nm. Changes in AMP concentrations of 5 μM or more can be reliably detected. The ease of use and relatively low expense make the AMP assay suitable for both high-throughput screening and kinetic analyses. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. Quantitative and Cost Comparison of Ultrasensitive Human Immunodeficiency Virus Type 1 RNA Viral Load Assays: Bayer bDNA Quantiplex Versions 3.0 and 2.0 and Roche PCR Amplicor Monitor Version 1.5

    PubMed Central

    Elbeik, Tarek; Charlebois, Edwin; Nassos, Patricia; Kahn, James; Hecht, Frederick M.; Yajko, David; Ng, Valerie; Hadley, Keith

    2000-01-01

    Quantification of human immunodeficiency virus type 1 (HIV-1) RNA as a measure of viral load has greatly improved the monitoring of therapies for infected individuals. With the significant reductions in viral load now observed in individuals treated with highly active anti-retroviral therapy (HAART), viral load assays have been adapted to achieve greater sensitivity. Two commercially available ultrasensitive assays, the Bayer Quantiplex HIV-1 bDNA version 3.0 (bDNA 3.0) assay and the Roche Amplicor HIV-1 Monitor Ultrasensitive version 1.5 (Amplicor 1.5) assay, are now being used to monitor HIV-1-infected individuals. Both of these ultrasensitive assays have a reported lower limit of 50 HIV-1 RNA copies/ml and were developed from corresponding older generation assays with lower limits of 400 to 500 copies/ml. However, the comparability of viral load data generated by these ultrasensitive assays and the relative costs of labor, disposables, and biohazardous wastes were not determined in most cases. In this study, we used matched clinical plasma samples to compare the quantification of the newer bDNA 3.0 assay with that of the older bDNA 2.0 assay and to compare the quantification and costs of the bDNA 3.0 assay and the Amplicor 1.5 assay. We found that quantification by the bDNA 3.0 assay was approximately twofold higher than that by the bDNA 2.0 assay and was highly correlated to that by the Amplicor 1.5 assay. Moreover, cost analysis based on labor, disposables, and biohazardous wastes showed significant savings with the bDNA 3.0 assay as compared to the costs of the Amplicor 1.5 assay. PMID:10699005

  13. Quantitative and cost comparison of ultrasensitive human immunodeficiency virus type 1 RNA viral load assays: Bayer bDNA quantiplex versions 3.0 and 2.0 and Roche PCR Amplicor monitor version 1.5.

    PubMed

    Elbeik, T; Charlebois, E; Nassos, P; Kahn, J; Hecht, F M; Yajko, D; Ng, V; Hadley, K

    2000-03-01

    Quantification of human immunodeficiency virus type 1 (HIV-1) RNA as a measure of viral load has greatly improved the monitoring of therapies for infected individuals. With the significant reductions in viral load now observed in individuals treated with highly active anti-retroviral therapy (HAART), viral load assays have been adapted to achieve greater sensitivity. Two commercially available ultrasensitive assays, the Bayer Quantiplex HIV-1 bDNA version 3.0 (bDNA 3.0) assay and the Roche Amplicor HIV-1 Monitor Ultrasensitive version 1.5 (Amplicor 1.5) assay, are now being used to monitor HIV-1-infected individuals. Both of these ultrasensitive assays have a reported lower limit of 50 HIV-1 RNA copies/ml and were developed from corresponding older generation assays with lower limits of 400 to 500 copies/ml. However, the comparability of viral load data generated by these ultrasensitive assays and the relative costs of labor, disposables, and biohazardous wastes were not determined in most cases. In this study, we used matched clinical plasma samples to compare the quantification of the newer bDNA 3.0 assay with that of the older bDNA 2.0 assay and to compare the quantification and costs of the bDNA 3.0 assay and the Amplicor 1.5 assay. We found that quantification by the bDNA 3.0 assay was approximately twofold higher than that by the bDNA 2.0 assay and was highly correlated to that by the Amplicor 1.5 assay. Moreover, cost analysis based on labor, disposables, and biohazardous wastes showed significant savings with the bDNA 3.0 assay as compared to the costs of the Amplicor 1.5 assay.

  14. Comparison of an assay using signal amplification of the heat-dissociated p24 antigen with the Roche Monitor human immunodeficiency virus RNA assay.

    PubMed

    Pascual, Alvaro; Cachafeiro, Ada; Funk, Michele L; Fiscus, Susan A

    2002-07-01

    We compared an assay using signal amplification of a heat-dissociated p24 antigen (HDAg) with the Roche Monitor human immunodeficiency virus (HIV) RNA assay. The two assays gave comparable results when 130 specimens from 130 patients were tested (r = 0.60, P < 0.0001). The HDAg assay was almost as sensitive (85%) as the Roche HIV RNA kit (95%), just as specific (25 negative results from 25 HIV seronegative volunteers [100%]), less variable (mean log standard deviation of 0.07 compared to 0.11 when eight specimens were tested three or four times), and less expensive (reagent and labor costs, $8 versus $75). The assay appeared to be useful for monitoring established patients (n = 17) and identifying seroconverters (n = 4). HIV subtypes A to F were all recognized. This assay should be useful for monitoring patients in resource-poor countries and for monitoring vaccine recipients.

  15. Monitoring environmental burden reduction from household waste prevention.

    PubMed

    Matsuda, Takeshi; Hirai, Yasuhiro; Asari, Misuzu; Yano, Junya; Miura, Takahiro; Ii, Ryota; Sakai, Shin-Ichi

    2018-01-01

    In this study, the amount of prevented household waste in Kyoto city was quantified using three methods. Subsequently, the greenhouse gas (GHG) emission reduction by waste prevention was calculated in order to monitor the impact of waste prevention. The methods of quantification were "relative change from baseline year (a)," "absolute change from potential waste generation (b)," and "absolute amount of activities (c)." Method (a) was popular for measuring waste prevention, but method (b) was the original approach to determine the absolute amount of waste prevention by estimating the potential waste generation. Method (c) also provided the absolute value utilizing the information of activities. Methods (b) and (c) enable the evaluation of the waste prevention activities with a similar baseline for recycling. Methods (b) and (c) gave significantly higher GHG reductions than method (a) because of the difference in baseline between them. Therefore, setting a baseline is very important for evaluating waste prevention. In practice, when focusing on the monitoring of a specific policy or campaign, method (a) is an appropriate option. On the other hand, when comparing the total impact of waste prevention to that of recycling, methods (b) and (c) should be applied. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Monitoring Acidophilic Microbes with Real-Time Polymerase Chain Reaction (PCR) Assays

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Frank F. Roberto

    2008-08-01

    Many techniques that are used to characterize and monitor microbial populations associated with sulfide mineral bioleaching require the cultivation of the organisms on solid or liquid media. Chemolithotrophic species, such as Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans, or thermophilic chemolithotrophs, such as Acidianus brierleyi and Sulfolobus solfataricus can grow quite slowly, requiring weeks to complete efforts to identify and quantify these microbes associated with bioleach samples. Real-time PCR (polymerase chain reaction) assays in which DNA targets are amplified in the presence of fluorescent oligonucleotide primers, allowing the monitoring and quantification of the amplification reactions as they progress, provide a means ofmore » rapidly detecting the presence of microbial species of interest, and their relative abundance in a sample. This presentation will describe the design and use of such assays to monitor acidophilic microbes in the environment and in bioleaching operations. These assays provide results within 2-3 hours, and can detect less than 100 individual microbial cells.« less

  17. Monitoring technologies for ocean disposal of radioactive waste

    NASA Astrophysics Data System (ADS)

    Triplett, M. B.; Solomon, K. A.; Bishop, C. B.; Tyce, R. C.

    1982-01-01

    The feasibility of using carefully selected subseabed locations to permanently isolate high level radioactive wastes at ocean depths greater than 4000 meters is discussed. Disposal at several candidate subseabed areas is being studied because of the long term geologic stability of the sediments, remoteness from human activity, and lack of useful natural resources. While the deep sea environment is remote, it also poses some significant challenges for the technology required to survey and monitor these sites, to identify and pinpoint container leakage should it occur, and to provide the environmental information and data base essential to determining the probable impacts of any such occurrence. Objectives and technical approaches to aid in the selective development of advanced technologies for the future monitoring of nuclear low level and high level waste disposal in the deep seabed are presented. Detailed recommendations for measurement and sampling technology development needed for deep seabed nuclear waste monitoring are also presented.

  18. Apparatus and method for quantitative assay of samples of transuranic waste contained in barrels in the presence of matrix material

    DOEpatents

    Caldwell, J.T.; Herrera, G.C.; Hastings, R.D.; Shunk, E.R.; Kunz, W.E.

    1987-08-28

    Apparatus and method for performing corrections for matrix material effects on the neutron measurements generated from analysis of transuranic waste drums using the differential-dieaway technique. By measuring the absorption index and the moderator index for a particular drum, correction factors can be determined for the effects of matrix materials on the ''observed'' quantity of fissile and fertile material present therein in order to determine the actual assays thereof. A barrel flux monitor is introduced into the measurement chamber to accomplish these measurements as a new contribution to the differential-dieaway technology. 9 figs.

  19. Development of an enzyme-linked immunosorbent assay and a beta-1 adrenergic receptor-based assay for monitoring the drug atenolol.

    PubMed

    Sapir, A; Shalev, A Hariton; Skalka, N; Bronshtein, A; Altstein, M

    2013-03-01

    Two approaches for monitoring atenolol (ATL) were applied: an immunochemical assay and a competitive-binding assay, based on the interaction between ATL and its target receptor, β1 adrenergic receptor (β1AR). Polyclonal antibodies (Abs) for ATL were generated, and a highly specific microplate immunochemical assay, that is, an enzyme-linked immunosorbent assay (ELISA), for its detection was developed. The ATL ELISA exhibited I50 and limit of detection (I20) values of 0.15 ± 0.048 and 0.032 ± 0.016 ng/ml, respectively, and the Abs did not cross-react with any of the tested beta-blocker drugs. Furthermore, a human β1AR (h-β1AR) was stably expressed in Spodoptera frugiperda cells (Sf9). The receptor was employed to develop a competitive-binding assay that monitored binding of ATL in the presence of isoproteranol by quantification of secondary messenger, cyclic adenosine monophosphate (cAMP), levels in the transfected cells. The assay showed that the recombinant h-β1AR was functional, could bind the agonistic ligand isoproterenol as well as the antagonist ATL, as indicated by a dose-dependent elevation of cAMP in the presence of isoproteranol, and decrease after ATL addition. The highly efficient and sensitive ELISA and the receptor assay represent two methods suitable for efficient and cost-effective large-scale, high-throughput monitoring of ATL in environmental, agricultural, and biological samples. Copyright © 2012 SETAC.

  20. A bioluminescent caspase-1 activity assay rapidly monitors inflammasome activation in cells.

    PubMed

    O'Brien, Martha; Moehring, Danielle; Muñoz-Planillo, Raúl; Núñez, Gabriel; Callaway, Justin; Ting, Jenny; Scurria, Mike; Ugo, Tim; Bernad, Laurent; Cali, James; Lazar, Dan

    2017-08-01

    Inflammasomes are protein complexes induced by diverse inflammatory stimuli that activate caspase-1, resulting in the processing and release of cytokines, IL-1β and IL-18, and pyroptosis, an immunogenic form of cell death. To provide a homogeneous method for detecting caspase-1 activity, we developed a bioluminescent, plate-based assay that combines a substrate, Z-WEHD-aminoluciferin, with a thermostable luciferase in an optimized lytic reagent added directly to cultured cells. Assay specificity for caspase-1 is conferred by inclusion of a proteasome inhibitor in the lytic reagent and by use of a caspase-1 inhibitor to confirm activity. This approach enables a specific and rapid determination of caspase-1 activation. Caspase-1 activity is stable in the reagent thereby providing assay convenience and flexibility. Using this assay system, caspase-1 activation has been determined in THP-1 cells following treatment with α-hemolysin, LPS, nigericin, gramicidin, MSU, R848, Pam3CSK4, and flagellin. Caspase-1 activation has also been demonstrated in treated J774A.1 mouse macrophages, bone marrow-derived macrophages (BMDMs) from mice, as well as in human primary monocytes. Caspase-1 activity was not detected in treated BMDMs derived from Casp1 -/- mice, further confirming the specificity of the assay. Caspase-1 activity can be measured directly in cultured cells using the lytic reagent, or caspase-1 activity released into medium can be monitored by assay of transferred supernatant. The caspase-1 assay can be multiplexed with other assays to monitor additional parameters from the same cells, such as IL-1β release or cell death. The caspase-1 assay in combination with a sensitive real-time monitor of cell death allows one to accurately establish pyroptosis. This assay system provides a rapid, convenient, and flexible method to specifically and quantitatively monitor caspase-1 activation in cells in a plate-based format. This will allow a more efficient and effective

  1. Developing and Evaluating Prototype of Waste Volume Monitoring Using Internet of Things

    NASA Astrophysics Data System (ADS)

    Fathhan Arief, Mohamad; Lumban Gaol, Ford

    2017-06-01

    In Indonesia, especially Jakarta have a lot of garbage strewn that can be an eyesore and also cause pollution that can carry diseases. Garbage strewn can cause many things, one of her dues is bins are overflowing due to the full so it can not accommodate the waste dumped from other people. Thus, the author created a new method for waste disposal more systematic. In creating new method requires a technology to supports, then the author makes a prototype for waste volume monitoring. By using the internet of things prototype of waste volume monitoring may give notification to the sanitary agency that waste in the trash bin needs to be disposal. In this study, conducted the design and manufactured of prototype waste volume monitoring using LinkItONE board based by Arduino and an ultrasonic sensor for appliance senses. Once the prototype is completed, evaluation in order to determine whether the prototype will function properly. The result showed that the expected function of a prototype waste volume monitoring can work well.

  2. Comparison of solid-phase and eluate assays to gauge the ecotoxicological risk of organic wastes on soil organisms.

    PubMed

    Domene, Xavier; Alcañiz, Josep M; Andrés, Pilar

    2008-02-01

    Development of methodologies to assess the safety of reusing polluted organic wastes in soil is a priority in Europe. In this study, and coupled with chemical analysis, seven organic wastes were subjected to different aquatic and soil bioassays. Tests were carried out with solid-phase waste and three different waste eluates (water, methanol, and dichloromethane). Solid-phase assays were indicated as the most suitable for waste testing not only in terms of relevance for real situations, but also because toxicity in eluates was generally not representative of the chronic effects in solid-phase. No general correlations were found between toxicity and waste pollutant burden, neither in solid-phase nor in eluate assays, showing the inability of chemical methods to predict the ecotoxicological risks of wastes. On the contrary, several physicochemical parameters reflecting the degree of low organic matter stability in wastes were the main contributors to the acute toxicity seen in collembolans and daphnids.

  3. 10 CFR 60.143 - Monitoring and testing waste packages.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... package monitoring program shall include laboratory experiments which focus on the internal condition of... the laboratory experiments. (d) The waste package monitoring program shall continue as long as...

  4. 10 CFR 60.143 - Monitoring and testing waste packages.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... package monitoring program shall include laboratory experiments which focus on the internal condition of... the laboratory experiments. (d) The waste package monitoring program shall continue as long as...

  5. 10 CFR 60.143 - Monitoring and testing waste packages.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... package monitoring program shall include laboratory experiments which focus on the internal condition of... the laboratory experiments. (d) The waste package monitoring program shall continue as long as...

  6. 10 CFR 60.143 - Monitoring and testing waste packages.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... package monitoring program shall include laboratory experiments which focus on the internal condition of... the laboratory experiments. (d) The waste package monitoring program shall continue as long as...

  7. 10 CFR 60.143 - Monitoring and testing waste packages.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... package monitoring program shall include laboratory experiments which focus on the internal condition of... the laboratory experiments. (d) The waste package monitoring program shall continue as long as...

  8. A simple assay system to monitor the potential for contamination during different stages of cryopreservation.

    PubMed

    Morris, G J

    2009-01-01

    A simple assay to monitor the potential for contamination during different steps of cryopreservation is described. The assay is based on the contamination of liquid nitrogen using crystals of sucrose hemi-heptahydrate, these are stable in liquid nitrogen, nitrogen vapour and ambient air and can be monitored by a simple assay which allows contamination risks to be evaluated in a direct, rapid manner.

  9. Screening complex hazardous wastes for mutagenic activity using a modified version of the TLC/Salmonella assay.

    PubMed

    Houk, V S; Claxton, L D

    1986-03-01

    10 complex hazardous wastes were tested for mutagenic activity using a modified version of the TLC/Salmonella assay developed by Bjørseth et al. (1982). This fractionation/bioassay scheme couples thin-layer chromatography (TLC) with the Salmonella/mammalian-microsome (Ames) assay for the detection of mutagenic constituents in complex mixtures. Crude (unadulterated) hazardous wastes and selected hazardous waste extracts were fractionated on commercially available cellulose TLC plates. Mutagenicity testing was performed in situ by applying a single overlay of minimal growth agar, tester strain TA98 or TA100, and the optional metabolic activation system directly onto the developed chromatogram. A mutagenic effect was indicated either by the appearance of localized clusters of revertant colonies or by an increase in total revertant growth vis-à-vis control plates. 7 of 10 hazardous wastes (including tars, emulsions, sludges, and spent acids and caustics) demonstrated mutagenic activity when tested by this method. To assess the sensitivity of the modified TLC/Salmonella assay, 14 Salmonella mutagens from a wide range of chemical classes and polarities were tested. Selected compounds included heterocyclics, aromatic amines, alkylating agents, antitumor agents, a nitrosamine and a nitroaromatic. 11 of the 14 mutagens were positive in this test system. The 3 compounds refractory to analysis included a polycyclic aromatic hydrocarbon and two volatiles.

  10. Are Fish and Standardized FETAX Assays Protective Enough for Amphibians? A Case Study on Xenopus laevis Larvae Assay with Biologically Active Substances Present in Livestock Wastes

    PubMed Central

    Martini, Federica; Tarazona, José V.; Pablos, M. Victoria

    2012-01-01

    Biologically active substances could reach the aquatic compartment when livestock wastes are considered for recycling. Recently, the standardized FETAX assay has been questioned, and some researchers have considered that the risk assessment performed on fish could not be protective enough to cover amphibians. In the present study a Xenopus laevis acute assay was developed in order to compare the sensitivity of larvae relative to fish or FETAX assays; veterinary medicines (ivermectin, oxytetracycline, tetracycline, sulfamethoxazole, and trimethoprim) and essential metals (zinc, copper, manganese, and selenium) that may be found in livestock wastes were used for the larvae exposure. Lethal (LC50) and sublethal effects were estimated. Available data in both, fish and FETAX studies, were in general more protective than values found out in the current study, but not in all cases. Moreover, the presence of nonlethal effects, caused by ivermectin, zinc, and copper, suggested that several physiological mechanisms could be affected. Thus, this kind of effects should be deeply investigated. The results obtained in the present study could expand the information about micropollutants from livestock wastes on amphibians. PMID:22629159

  11. Use of theoretical waste inventories in planning and monitoring of hazardous waste management systems.

    PubMed

    Yilmaz, Ozge; Can, Zehra S; Toroz, Ismail; Dogan, Ozgur; Oncel, Salim; Alp, Emre; Dilek, Filiz B; Karanfil, Tanju; Yetis, Ulku

    2014-08-01

    Hazardous waste (HW) generation information is an absolute necessity for ensuring the proper planning, implementation, and monitoring of any waste management system. Unfortunately, environmental agencies in developing countries face difficulties in gathering data directly from the creators of such wastes. It is possible, however, to construct theoretical HW inventories using the waste generation factors (WGFs). The objective of this study was to develop a complete nationwide HW inventory of Turkey that relies on nation-specific WGFs to support management activities of the Turkish Ministry of Environment and Urbanization (MoEU). Inventory studies relied on WGFs from: (a) the literature and (b) field studies and analysis of waste declarations reflecting country-specific industrial practices. Moreover, new tools were introduced to the monitoring infrastructure of MoEU to obtain a comprehensive waste generation data set. Through field studies and a consideration of country specific conditions, it was possible to more thoroughly elucidate HW generation trends in Turkey, a method that was deemed superior to other alternatives. Declaration and literature based WGFs also proved most helpful in supplementing field observations that could not always be conducted. It was determined that these theoretical inventories could become valuable assets in supporting regulating agencies in developing countries for a more thorough implementation of HW management systems. © The Author(s) 2014.

  12. Recovering lead from cupel waste generated in gold analysis by Pb-Fire assay.

    PubMed

    Cerceau, Cristiane Isaac; Carvalho, Cornélio de Freitas; Rabelo, Ana Carolina Silveira; Dos Santos, Cláudio Gouvea; Gonçalves, Sabrina Mayra Dias; Varejão, Eduardo Vinícius Vieira

    2016-12-01

    Because of its precision and accuracy, Pb-Fire assay is the most employed method for gold analysis in geological materials. At the second stage of the method, namely cupellation, lead is oxidized to PbO which is absorbed by the cupel, leading to metallic gold as a tiny bend at the bottom of the recipient. After cupellation, cupel becomes highly contaminated with lead, making its disposal a serious risk of environmental contamination. In the present work, a leaching process for removing lead from cupel waste is proposed, which allowed for removing 96% of PbO by weight. After a precipitation step, 92.0% of lead was recovered from leachates in the form of PbSO 4 . Lead in the solid wastes left by the extraction was above the limit established by Brazilian legislation and these were classified as non-hazardous. Finally, secondary effluents generated after the precipitation step presented lead content more than twenty times lower than that of leachates from cupel waste. Tons of cupel waste are annually generated from gold analysis by Pb-Fire assay. Thus, the proposed method can contribute to prevent the discharge of high amounts of lead into the environment. Also, recovery of lead can help to partially meet the industrial demand for lead compounds. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. Quantitative CrAssphage PCR Assays for Human Fecal ...

    EPA Pesticide Factsheets

    Environmental waters are monitored for fecal pollution to protect public health and water resources. Traditionally, general fecal indicator bacteria are used; however, they cannot distinguish human fecal waste from pollution from other animals. Recently, a novel bacteriophage, crAssphage, was discovered by metagenomic data mining and reported to be abundant in and closely associated with human fecal waste. To confirm bioinformatic predictions, 384 primer sets were designed along the length of the crAssphage genome. Based upon initial screening, two novel crAssphage qPCR assays (CPQ_056 and CPQ_064) were designed and evaluated in reference fecal samples and water matrices. The assays exhibited high specificities (98.6%) when tested against a large animal fecal reference library and were highly abundant in raw sewage and sewage impacted water samples. In addition, CPQ_056 and CPQ_064 assay performance was compared to HF183/BacR287 and HumM2 methods in paired experiments. Findings confirm viral crAssphage qPCR assays perform at a similar level to well established bacterial human-associated fecal source identification technologies. These new viral based assays could become important water quality management and research tools. To inform the public.

  14. Real-time alpha monitoring of a radioactive liquid waste stream at Los Alamos National Laboratory

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Johnson, J.D.; Whitley, C.R.; Rawool-Sullivan, M.

    1995-12-31

    This poster display concerns the development, installation, and testing of a real-time radioactive liquid waste monitor at Los Alamos National Laboratory (LANL). The detector system was designed for the LANL Radioactive Liquid Waste Treatment Facility so that influent to the plant could be monitored in real time. By knowing the activity of the influent, plant operators can better monitor treatment, better segregate waste (potentially), and monitor the regulatory compliance of users of the LANL Radioactive Liquid Waste Collection System. The detector system uses long-range alpha detection technology, which is a nonintrusive method of characterization that determines alpha activity on themore » liquid surface by measuring the ionization of ambient air. Extensive testing has been performed to ensure long-term use with a minimal amount of maintenance. The final design was a simple cost-effective alpha monitor that could be modified for monitoring influent waste streams at various points in the LANL Radioactive Liquid Waste Collection System.« less

  15. Development of a Simple Dipstick Assay for Operational Monitoring of DDT.

    PubMed

    Ismail, Hanafy M; Kumar, Vijay; Singh, Rudra P; Williams, Christopher; Shivam, Pushkar; Ghosh, Ayan; Deb, Rinki; Foster, Geraldine M; Hemingway, Janet; Coleman, Michael; Coleman, Marlize; Das, Pradeep; Paine, Mark J I

    2016-01-01

    Indoor residual spraying (IRS) of DDT is used to control visceral leishmaniasis (VL) in India. However, the quality of spraying is severely compromised by a lack of affordable field assays to monitor target doses of insecticide. Our aim was to develop a simple DDT insecticide quantification kit (IQK) for monitoring DDT levels in an operational setting. DDT quantification was based on the stoichiometric release of chloride from DDT by alkaline hydrolysis and detection of the released ion using Quantab chloride detection strips. The assay was specific for insecticidal p,p`-DDT (LoQ = 0.082 g/m2). Bostik discs were effective in post spray wall sampling, extracting 25-70% of active ingredient depending on surface. Residual DDT was sampled from walls in Bihar state in India using Bostik adhesive discs and DDT concentrations (g p,p`-DDT/m2) were determined using IQK and HPLC (n = 1964 field samples). Analysis of 161 Bostik samples (pooled sample pairs) by IQK and HPLC produced excellent correlation (R2 = 0.96; Bland-Altman bias = -0.0038). IQK analysis of the remaining field samples matched HPLC data in identifying households that had been under sprayed, in range or over sprayed. A simple dipstick assay has been developed for monitoring DDT spraying that gives comparable results to HPLC. By making laboratory-based analysis of DDT dosing accessible to field operatives, routine monitoring of DDT levels can be promoted in low- and middle- income countries to maximise the effectiveness of IRS.

  16. Shuttle era waste management and biowaste monitoring

    NASA Technical Reports Server (NTRS)

    Sauer, R. L.; Fogal, G. L.

    1976-01-01

    The acquisition of crew biomedical data has been an important task on manned space missions. The monitoring of biowastes from the crew to support water and mineral balance studies and endocrine studies has been a valuable part of this activity. This paper will present a review of waste management systems used in past programs. This past experience will be cited as to its influence on the Shuttle design. Finally, the Shuttle baseline waste management system and the proposed Shuttle biomedical measurement and sampling systems will be presented.

  17. In-line assay monitor for uranium hexafluoride

    DOEpatents

    Wallace, S.A.

    1980-03-21

    An in-line assay monitor for determining the content of uranium-235 in a uranium hexafluoride gas isotopic separation system is provided which removes the necessity of complete access to the operating parameters of the system for determining the uranium-235 content. The method and monitor for carrying out the method involve cooling of a radiation pervious chamber connected in fluid communication with the selected point in the system to withdraw a specimen and solidify the specimen in the chamber. The specimen is irradiated by means of an ionizing radiation source of energy different from that of the 185 keV gamma emissions from uranium-235. The uranium-235 content of the specimen is determined from comparison of the accumulated 185 keV energy counts and reference energy counts. The latter is used to measure the total uranium isotopic content of the specimen.

  18. Development of a Simple Dipstick Assay for Operational Monitoring of DDT

    PubMed Central

    Ismail, Hanafy M.; Kumar, Vijay; Singh, Rudra P.; Williams, Christopher; Shivam, Pushkar; Ghosh, Ayan; Deb, Rinki; Foster, Geraldine M.; Hemingway, Janet; Coleman, Michael; Coleman, Marlize; Das, Pradeep; Paine, Mark J. I.

    2016-01-01

    Background Indoor residual spraying (IRS) of DDT is used to control visceral leishmaniasis (VL) in India. However, the quality of spraying is severely compromised by a lack of affordable field assays to monitor target doses of insecticide. Our aim was to develop a simple DDT insecticide quantification kit (IQK) for monitoring DDT levels in an operational setting. Methodology/ principle findings DDT quantification was based on the stoichiometric release of chloride from DDT by alkaline hydrolysis and detection of the released ion using Quantab chloride detection strips. The assay was specific for insecticidal p,p`-DDT (LoQ = 0.082 g/m2). Bostik discs were effective in post spray wall sampling, extracting 25–70% of active ingredient depending on surface. Residual DDT was sampled from walls in Bihar state in India using Bostik adhesive discs and DDT concentrations (g p,p`-DDT/m2) were determined using IQK and HPLC (n = 1964 field samples). Analysis of 161 Bostik samples (pooled sample pairs) by IQK and HPLC produced excellent correlation (R2 = 0.96; Bland-Altman bias = −0.0038). IQK analysis of the remaining field samples matched HPLC data in identifying households that had been under sprayed, in range or over sprayed. Interpretation A simple dipstick assay has been developed for monitoring DDT spraying that gives comparable results to HPLC. By making laboratory-based analysis of DDT dosing accessible to field operatives, routine monitoring of DDT levels can be promoted in low- and middle- income countries to maximise the effectiveness of IRS. PMID:26760773

  19. Design and application of a fluorogenic assay for monitoring inflammatory caspase activity.

    PubMed

    Ranganathan, Raj; Lenti, Gena; Tassone, Nicholas M; Scannell, Brian J; Southern, Cathrine A; Karver, Caitlin E

    2018-02-15

    Various fluorogenic assays exist for monitoring the activity of inflammatory caspases. However, there are no continuous assays that provide C-terminal substrate sequence specificity for inflammatory caspases. As a first step towards this, we have developed a continuous in vitro assay that relies on monitoring emission from tryptophan after cleavage of a quenching coumarin chromophore. The coumarin can be attached as an amino acid side chain or capping the C-terminus of the peptide. When the coumarin is a side chain, it allows for C-terminal and N-terminal sequence specificities to be explored. Using this assay, we obtained Michaelis-Menten kinetic data for four proof-of-principle peptides: WEHD-AMC (K M  = 15 ± 2 μM), WEHD-MCA (K M  = 93 ± 19 μM), WEHDG-MCA (K M  = 21 ± 6 μM) and WEHDA-MCA (K M  = 151 ± 37 μM), where AMC is 7-amino-4-methylcoumarin and MCA is β-(7-methoxy-coumarin-4-yl)-Ala. The results indicate the viability of this new assay approach in the design of effective fluorogenic substrates for inflammatory caspases. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Kinetic Tetrazolium Microtiter Assay

    NASA Technical Reports Server (NTRS)

    Pierson, Duane L.; Stowe, Raymond; Koenig, David

    1993-01-01

    Kinetic tetrazolium microtiter assay (KTMA) involves use of tetrazolium salts and Triton X-100 (or equivalent), nontoxic, in vitro color developer solubilizing colored metabolite formazan without injuring or killing metabolizing cells. Provides for continuous measurement of metabolism and makes possible to determine rate of action of antimicrobial agent in real time as well as determines effective inhibitory concentrations. Used to monitor growth after addition of stimulatory compounds. Provides for kinetic determination of efficacy of biocide, greatly increasing reliability and precision of results. Also used to determine relative effectiveness of antimicrobial agent as function of time. Capability of generating results on day of test extremely important in treatment of water and waste, disinfection of hospital rooms, and in pharmaceutical, agricultural, and food-processing industries. Assay also used in many aspects of cell biology.

  1. An eDNA Assay to Monitor a Globally Invasive Fish Species from Flowing Freshwater.

    PubMed

    Adrian-Kalchhauser, Irene; Burkhardt-Holm, Patricia

    2016-01-01

    Ponto-Caspian gobies are a flock of five invasive fish species that have colonized freshwaters and brackish waters in Europe and North America. One of them, the round goby Neogobius melanostomus, figures among the 100 worst invaders in Europe. Current methods to detect the presence of Ponto-Caspian gobies involve catching or sighting the fish. These approaches are labor intense and not very sensitive. Consequently, populations are usually detected only when they have reached high densities and when management or containment efforts are futile. To improve monitoring, we developed an assay based on the detection of DNA traces (environmental DNA, or eDNA) of Ponto-Caspian gobies in river water. The assay specifically detects invasive goby DNA and does not react to any native fish species. We apply the assay to environmental samples and demonstrate that parameters such as sampling depth, sampling location, extraction protocol, PCR protocol and PCR inhibition greatly impact detection. We further successfully outline the invasion front of Ponto-Caspian gobies in a large river, the High Rhine in Switzerland, and thus demonstrate the applicability of the assay to lotic environments. The eDNA assay requires less time, equipment, manpower, skills, and financial resources than the conventional monitoring methods such as electrofishing, angling or diving. Samples can be taken by untrained individuals, and the assay can be performed by any molecular biologist on a conventional PCR machine. Therefore, this assay enables environment managers to map invaded areas independently of fishermen's' reports and fish community monitorings.

  2. An eDNA Assay to Monitor a Globally Invasive Fish Species from Flowing Freshwater

    PubMed Central

    Adrian-Kalchhauser, Irene; Burkhardt-Holm, Patricia

    2016-01-01

    Ponto-Caspian gobies are a flock of five invasive fish species that have colonized freshwaters and brackish waters in Europe and North America. One of them, the round goby Neogobius melanostomus, figures among the 100 worst invaders in Europe. Current methods to detect the presence of Ponto-Caspian gobies involve catching or sighting the fish. These approaches are labor intense and not very sensitive. Consequently, populations are usually detected only when they have reached high densities and when management or containment efforts are futile. To improve monitoring, we developed an assay based on the detection of DNA traces (environmental DNA, or eDNA) of Ponto-Caspian gobies in river water. The assay specifically detects invasive goby DNA and does not react to any native fish species. We apply the assay to environmental samples and demonstrate that parameters such as sampling depth, sampling location, extraction protocol, PCR protocol and PCR inhibition greatly impact detection. We further successfully outline the invasion front of Ponto-Caspian gobies in a large river, the High Rhine in Switzerland, and thus demonstrate the applicability of the assay to lotic environments. The eDNA assay requires less time, equipment, manpower, skills, and financial resources than the conventional monitoring methods such as electrofishing, angling or diving. Samples can be taken by untrained individuals, and the assay can be performed by any molecular biologist on a conventional PCR machine. Therefore, this assay enables environment managers to map invaded areas independently of fishermen’s’ reports and fish community monitorings. PMID:26814998

  3. The help of simulation codes in designing waste assay systems using neutron measurement methods: Application to the alpha low level waste assay system PROMETHEE 6

    NASA Astrophysics Data System (ADS)

    Mariani, A.; Passard, C.; Jallu, F.; Toubon, H.

    2003-11-01

    The design of a specific nuclear assay system for a dedicated application begins with a phase of development, which relies on information from the literature or on knowledge resulting from experience, and on specific experimental verifications. The latter ones may require experimental devices which can be restricting in terms of deadline, cost and safety. One way generally chosen to bypass these difficulties is to use simulation codes to study particular aspects. This paper deals with the potentialities offered by the simulation in the case of a passive-active neutron (PAN) assay system for alpha low level waste characterization; this system has been carried out at the Nuclear Measurements Development Laboratory of the French Atomic Energy Commission. Due to the high number of parameters to be taken into account for its development, this is a particularly sophisticated example. Since the PAN assay system, called PROMETHEE (prompt epithermal and thermal interrogation experiment), must have a detection efficiency of more than 20% and preserve a high level of modularity for various applications, an improved version has been studied using the MCNP4 (Monte Carlo N-Particle) transport code. Parameters such as the dimensions of the assay system, of the cavity and of the detection blocks, and the thicknesses of the nuclear materials of neutronic interest have been optimised. Therefore, the number of necessary experiments was reduced.

  4. Characterization of temporal variations in landfill gas components inside an open solid waste dump site in Sri Lanka.

    PubMed

    Nagamori, Masanao; Mowjood, M I M; Watanabe, Youichi; Isobe, Yugo; Ishigaki, Tomonori; Kawamoto, Ken

    2016-12-01

    A long-term monitoring of composition of landfill gases in the region with high rainfall was conducted using an argon assay in order to discuss air intrusion into the dump site. Gas samples were taken from vertical gas monitoring pipes installed along transects at two sections (called new and old) of an abandoned waste dump site in Sri Lanka. N 2 O concentrations varied especially widely, by more than three orders of magnitude (0.046-140 ppmv). The nitrogen/argon ratio of landfill gas was normally higher than that of fresh air, implying that denitrification occurred in the dump site. Argon assays indicate that both N 2 and N 2 O production occurred inside waste and more significantly in the old section. The Ar assay would help for evaluations of N 2 O emission in developing countries. A long-term monitoring of composition of landfill gases in the region with high rainfall was conducted using an argon assay in order to discuss air intrusion into the dump site. Argon assays indicate that both N 2 and N 2 O production occurred inside waste and more significantly in the old section.

  5. NRC Perspectives on Waste Incidental to Reprocessing Consultations and Monitoring - 13398

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    McKenney, Christepher A.; Suber, Gregory F.; Felsher, Harry D.

    2013-07-01

    Section 3116 of the Ronald W. Reagan National Defense Authorization Act for Fiscal Year 2005 (NDAA) requires the U.S. Department of Energy (DOE) to consult with the U.S. Nuclear Regulatory Commission (NRC) for certain non-high level waste (HLW) determinations. The NDAA also requires NRC to monitor DOE's disposal actions related to those determinations to assess compliance with NRC regulations in 10 CFR Part 61, Subpart C. The NDAA applies to DOE activities that will remain within the States of South Carolina and Idaho. DOE has chosen to, under DOE Order 435.1, engage in consultation with NRC for similar activities inmore » the State of Washington and New York, however, the NRC has no monitoring responsibilities. In 2007, the NRC developed a draft Final Report for Interim Use entitled, NUREG-1854: NRC Staff Guidance for Activities Related to U.S. Department of Energy Waste Determinations. Since the law was enacted, the DOE and NRC have consulted on three waste determinations within the affected States: (1) the Saltstone Disposal Facility at the Savannah River Site (SRS) within the State of South Carolina in 2005, (2) the INTEC Tank Farm at the Idaho National Laboratory within the State of Idaho in 2006, and (3) the F Tank Farm at SRS in 2011. After the end of consultation and issuance by DOE of the final waste determination, monitoring began at each of these sites, including the development of monitoring plans. In addition to the NDAA sites, DOE has requested NRC consultation support on both individual tanks and the entire C Tank Farm at the Hanford Nuclear Reservation in the State of Washington. DOE also requested consultation of waste determinations performed on the melter and related feed tanks at the West Valley site in New York that would be disposed offsite. In the next few years, NRC and DOE will consult on the last of the NDAA waste determinations for a while, the H Tank Farm waste determination at SRS. DOE may identify other activities in the future but

  6. Iraq liquid radioactive waste tanks maintenance and monitoring program plan.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dennis, Matthew L.; Cochran, John Russell; Sol Shamsaldin, Emad

    2011-10-01

    The purpose of this report is to develop a project management plan for maintaining and monitoring liquid radioactive waste tanks at Iraq's Al-Tuwaitha Nuclear Research Center. Based on information from several sources, the Al-Tuwaitha site has approximately 30 waste tanks that contain varying amounts of liquid or sludge radioactive waste. All of the tanks have been non-operational for over 20 years and most have limited characterization. The program plan embodied in this document provides guidance on conducting radiological surveys, posting radiation control areas and controlling access, performing tank hazard assessments to remove debris and gain access, and conducting routine tankmore » inspections. This program plan provides general advice on how to sample and characterize tank contents, and how to prioritize tanks for soil sampling and borehole monitoring.« less

  7. Bremsstrahlung-Based Imaging and Assays of Radioactive, Mixed and Hazardous Waste

    NASA Astrophysics Data System (ADS)

    Kwofie, J.; Wells, D. P.; Selim, F. A.; Harmon, F.; Duttagupta, S. P.; Jones, J. L.; White, T.; Roney, T.

    2003-08-01

    A new nondestructive accelerator based x-ray fluorescence (AXRF) approach has been developed to identify heavy metals in large-volume samples. Such samples are an important part of the process and waste streams of U.S Department of Energy sites, as well as other industries such as mining and milling. Distributions of heavy metal impurities in these process and waste samples can range from homogeneous to highly inhomogeneous, and non-destructive assays and imaging that can address both are urgently needed. Our approach is based on using high-energy, pulsed bremsstrahlung beams (3-6.5 MeV) from small electron accelerators to produce K-shell atomic fluorescence x-rays. In addition we exploit pair-production, Compton scattering and x-ray transmission measurements from these beams to probe locations of high density and high atomic number. The excellent penetrability of these beams allows assays and images for soil-like samples at least 15 g/cm2 thick, with elemental impurities of atomic number greater than approximately 50. Fluorescence yield of a variety of targets was measured as a function of impurity atomic number, impurity homogeneity, and sample thickness. We report on actual and potential detection limits of heavy metal impurities in a soil matrix for a variety of samples, and on the potential for imaging, using AXRF and these related probes.

  8. Integrated technologies for solid waste bin monitoring system.

    PubMed

    Arebey, Maher; Hannan, M A; Basri, Hassan; Begum, R A; Abdullah, Huda

    2011-06-01

    The integration of communication technologies such as radio frequency identification (RFID), global positioning system (GPS), general packet radio system (GPRS), and geographic information system (GIS) with a camera are constructed for solid waste monitoring system. The aim is to improve the way of responding to customer's inquiry and emergency cases and estimate the solid waste amount without any involvement of the truck driver. The proposed system consists of RFID tag mounted on the bin, RFID reader as in truck, GPRS/GSM as web server, and GIS as map server, database server, and control server. The tracking devices mounted in the trucks collect location information in real time via the GPS. This information is transferred continuously through GPRS to a central database. The users are able to view the current location of each truck in the collection stage via a web-based application and thereby manage the fleet. The trucks positions and trash bin information are displayed on a digital map, which is made available by a map server. Thus, the solid waste of the bin and the truck are being monitored using the developed system.

  9. Assessing and monitoring soil quality at agricultural waste disposal areas-Soil Indicators

    NASA Astrophysics Data System (ADS)

    Doula, Maria; Kavvadias, Victor; Sarris, Apostolos; Lolos, Polykarpos; Liakopoulou, Nektaria; Hliaoutakis, Aggelos; Kydonakis, Aris

    2014-05-01

    The necessity of elaborating indicators is one of the priorities identified by the United Nations Convention to Combat Desertification (UNCCD). The establishment of an indicator monitoring system for environmental purposes is dependent on the geographical scale. Some indicators such as rain seasonality or drainage density are useful over large areas, but others such as soil depth, vegetation cover type, and land ownership are only applicable locally. In order to practically enhance the sustainability of land management, research on using indicators for assessing land degradation risk must initially focus at local level because management decisions by individual land users are taken at this level. Soils that accept wastes disposal, apart from progressive degradation, may cause serious problems to the surrounding environment (humans, animals, plants, water systems, etc.), and thus, soil quality should be necessarily monitored. Therefore, quality indicators, representative of the specific waste type, should be established and monitored periodically. Since waste composition is dependent on their origin, specific indicators for each waste type should be established. Considering agricultural wastes, such a specification, however, could be difficult, since almost all agricultural wastes are characterized by increased concentrations of the same elements, namely, phosphorous, nitrogen, potassium, sulfur, etc.; contain large amounts of organic matter; and have very high values of chemical oxygen demand (COD), biochemical oxygen demand (BOD), and electrical conductivity. Two LIFE projects, namely AgroStrat and PROSODOL are focused on the identification of soil indicators for the assessment of soil quality at areas where pistachio wastes and olive mill wastes are disposed, respectively. Many soil samples were collected periodically for 2 years during PROSODOL and one year during AgroStrat (this project is in progress) from waste disposal areas and analyzed for 23 parameters

  10. 2014 Radiological Monitoring Results Associated with the Advanced Test Reactor Complex Cold Waste Pond

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lewis, Mike

    2015-02-01

    This report summarizes radiological monitoring performed of the Idaho National Laboratory Site’s Advanced Test Reactor Complex Cold Waste wastewater prior to discharge into the Cold Waste Pond and of specific groundwater monitoring wells associated with the Industrial Wastewater Reuse Permit (#LA-000161-01, Modification B). All radiological monitoring is performed to fulfill Department of Energy requirements under the Atomic Energy Act.

  11. 40 CFR 60.1320 - How do I monitor the load of my municipal waste combustion unit?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... municipal waste combustion unit? 60.1320 Section 60.1320 Protection of Environment ENVIRONMENTAL PROTECTION... of Performance for Small Municipal Waste Combustion Units for Which Construction is Commenced After... Monitoring Requirements § 60.1320 How do I monitor the load of my municipal waste combustion unit? (a) If...

  12. 40 CFR 60.1320 - How do I monitor the load of my municipal waste combustion unit?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... municipal waste combustion unit? 60.1320 Section 60.1320 Protection of Environment ENVIRONMENTAL PROTECTION... of Performance for Small Municipal Waste Combustion Units for Which Construction is Commenced After... Monitoring Requirements § 60.1320 How do I monitor the load of my municipal waste combustion unit? (a) If...

  13. 40 CFR 60.1320 - How do I monitor the load of my municipal waste combustion unit?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... municipal waste combustion unit? 60.1320 Section 60.1320 Protection of Environment ENVIRONMENTAL PROTECTION... of Performance for Small Municipal Waste Combustion Units for Which Construction is Commenced After... Monitoring Requirements § 60.1320 How do I monitor the load of my municipal waste combustion unit? (a) If...

  14. 40 CFR 60.1320 - How do I monitor the load of my municipal waste combustion unit?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... municipal waste combustion unit? 60.1320 Section 60.1320 Protection of Environment ENVIRONMENTAL PROTECTION... of Performance for Small Municipal Waste Combustion Units for Which Construction is Commenced After... Monitoring Requirements § 60.1320 How do I monitor the load of my municipal waste combustion unit? (a) If...

  15. 40 CFR 60.1320 - How do I monitor the load of my municipal waste combustion unit?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... municipal waste combustion unit? 60.1320 Section 60.1320 Protection of Environment ENVIRONMENTAL PROTECTION... of Performance for Small Municipal Waste Combustion Units for Which Construction is Commenced After... Monitoring Requirements § 60.1320 How do I monitor the load of my municipal waste combustion unit? (a) If...

  16. Ground Water Monitoring Requirements for Hazardous Waste Treatment, Storage and Disposal Facilities

    EPA Pesticide Factsheets

    The groundwater monitoring requirements for hazardous waste treatment, storage and disposal facilities (TSDFs) are just one aspect of the Resource Conservation and Recovery Act (RCRA) hazardous waste management strategy for protecting human health and the

  17. Early detection and evaluation of waste through sensorized containers for a collection monitoring application.

    PubMed

    Rovetta, Alberto; Xiumin, Fan; Vicentini, Federico; Minghua, Zhu; Giusti, Alessandro; Qichang, He

    2009-12-01

    The present study describes a novel application for use in the monitoring of municipal solid waste, based on distributed sensor technology and geographical information systems. Original field testing and evaluation of the application were carried out in Pudong, Shanghai (PR China). The local waste management system in Pudong features particular requirements related to the rapidly increasing rate of waste production. In view of the fact that collected waste is currently deployed to landfills or to incineration plants within the context investigated, the key aspects to be taken into account in waste collection procedures include monitoring of the overall amount of waste produced, quantitative measurement of the waste present at each collection point and identification of classes of material present in the collected waste. The case study described herein focuses particularly on the above mentioned aspects, proposing the implementation of a network of sensorized waste containers linked to a data management system. Containers used were equipped with a set of sensors mounted onto standard waste bins. The design, implementation and validation procedures applied are subsequently described. The main aim to be achieved by data collection and evaluation was to provide for feasibility analysis of the final device. Data pertaining to the content of waste containers, sampled and processed by means of devices validated on two purpose-designed prototypes, were therefore uploaded to a central monitoring server using GPRS connection. The data monitoring and management modules are integrated into an existing application used by local municipal authorities. A field test campaign was performed in the Pudong area. The system was evaluated in terms of real data flow from the network nodes (containers) as well as in terms of optimization functions, such as collection vehicle routing and scheduling. The most important outcomes obtained were related to calculations of waste weight and

  18. A Database of Reaction Monitoring Mass Spectrometry Assays for Elucidating Therapeutic Response in Cancer

    PubMed Central

    Remily-Wood, Elizabeth R.; Liu, Richard Z.; Xiang, Yun; Chen, Yi; Thomas, C. Eric; Rajyaguru, Neal; Kaufman, Laura M.; Ochoa, Joana E.; Hazlehurst, Lori; Pinilla-Ibarz, Javier; Lancet, Jeffrey; Zhang, Guolin; Haura, Eric; Shibata, David; Yeatman, Timothy; Smalley, Keiran S.M.; Dalton, William S.; Huang, Emina; Scott, Ed; Bloom, Gregory C.; Eschrich, Steven A.; Koomen, John M.

    2012-01-01

    Purpose The Quantitative Assay Database (QuAD), http://proteome.moffitt.org/QUAD/, facilitates widespread implementation of quantitative mass spectrometry in cancer biology and clinical research through sharing of methods and reagents for monitoring protein expression and modification. Experimental Design Liquid chromatography coupled to multiple reaction monitoring mass spectrometry (LC-MRM) assays are developed using SDS-PAGE fractionated lysates from cancer cell lines. Pathway maps created using GeneGO Metacore provide the biological relationships between proteins and illustrate concepts for multiplexed analysis; each protein can be selected to examine assay development at the protein and peptide level. Results The coupling of SDS-PAGE and LC-MRM screening has been used to detect 876 peptides from 218 cancer-related proteins in model systems including colon, lung, melanoma, leukemias, and myeloma, which has led to the development of 95 quantitative assays including stable-isotope labeled peptide standards. Methods are published online and peptide standards are made available to the research community. Protein expression measurements for heat shock proteins, including a comparison with ELISA and monitoring response to the HSP90 inhibitor, 17-DMAG, are used to illustrate the components of the QuAD and its potential utility. Conclusions and Clinical Relevance This resource enables quantitative assessment of protein components of signaling pathways and biological processes and holds promise for systematic investigation of treatment responses in cancer. PMID:21656910

  19. Comparative evaluation of hepatitis C virus RNA quantitation by branched DNA, NASBA, and monitor assays.

    PubMed

    Lunel, F; Cresta, P; Vitour, D; Payan, C; Dumont, B; Frangeul, L; Reboul, D; Brault, C; Piette, J C; Huraux, J M

    1999-02-01

    Several studies have shown a relationship between pretreatment hepatitis C virus (HCV) viral load and the response to interferon (IFN) therapy, creating a need for quantitative HCV-RNA assays. Here, we compared three commercial methods: nucleic acid sequence-based amplification NASBA (Organon), branched DNA 2.0 (bDNA) (Chiron), and Monitor (Roche), with reverse-transcription polymerase chain reaction (RT-PCR) as the reference. We assessed sensitivity and reproducibility on a well-characterized panel of sera (EUROHEP), a Chimp Rodney plasma pool, and samples from IFN-treated and -untreated patients with chronic hepatitis C caused by different HCV genotypes. The reproducibility of the NASBA and bDNA methods was slightly better than that of Monitor, especially for genotypes 2 and 4. NASBA had the highest sensitivity (99% vs. 94% and 88% with Monitor and bDNA, respectively), especially for the follow-up of patients on IFN. NASBA gave the highest HCV-RNA concentrations, which were approximately 10-fold more than with the bDNA assay and 100-fold more than with the Monitor kit. The linearity, tested on the chimp Rodney plasma pool, was better with bDNA for high viral load than with NASBA and Monitor, although for low concentration of HCV RNA, bDNA was negative. Pretreatment viral load was lower in patients who had a sustained virological response to IFN, although the bDNA method was not sensitive enough to quantify all pretreatment samples. This study indicates that gene amplification methods (NASBA or Monitor) have better sensitivity than bDNA assays for quantification of HCV RNA in patients with chronic HCV infection, although the bDNA and NASBA methods are more likely to quantify all genotypes. Prospective studies are needed to demonstrate the usefulness of quantitative assays for the follow-up of patients with chronic hepatitis C.

  20. In-line assay monitor for uranium hexafluoride

    DOEpatents

    Wallace, Steven A.

    1981-01-01

    An in-line assay monitor for determining the content of uranium-235 in a uranium hexafluoride gas isotopic separation system is provided which removes the necessity of complete access to the operating parameters of the system for determining the uranium-235 content. The monitor is intended for uses such as safeguard applications to assure that weapons grade uranium is not being produced in an enrichment cascade. The method and monitor for carrying out the method involve cooling of a radiation pervious chamber connected in fluid communication with the selected point in the system to withdraw a specimen and solidify the specimen in the chamber. The specimen is irradiated by means of an ionizing radiation source of energy different from that of the 185 keV gamma emissions from the uranium-235 present in the specimen. Simultaneously, the gamma emissions from the uranium-235 of the specimen and the source emissions transmitted through the sample are counted and stored in a multiple channel analyzer. The uranium-235 content of the specimen is determined from the comparison of the accumulated 185 keV energy counts and the reference energy counts. The latter is used to measure the total uranium isotopic content of the specimen. The process eliminates the necessity of knowing the system operating conditions and yet obtains the necessary data without need for large scintillation crystals and sophisticated mechanical designs.

  1. Radio Frequency Identification (RFID) and communication technologies for solid waste bin and truck monitoring system.

    PubMed

    Hannan, M A; Arebey, Maher; Begum, R A; Basri, Hassan

    2011-12-01

    This paper deals with a system of integration of Radio Frequency Identification (RFID) and communication technologies for solid waste bin and truck monitoring system. RFID, GPS, GPRS and GIS along with camera technologies have been integrated and developed the bin and truck intelligent monitoring system. A new kind of integrated theoretical framework, hardware architecture and interface algorithm has been introduced between the technologies for the successful implementation of the proposed system. In this system, bin and truck database have been developed such a way that the information of bin and truck ID, date and time of waste collection, bin status, amount of waste and bin and truck GPS coordinates etc. are complied and stored for monitoring and management activities. The results showed that the real-time image processing, histogram analysis, waste estimation and other bin information have been displayed in the GUI of the monitoring system. The real-time test and experimental results showed that the performance of the developed system was stable and satisfied the monitoring system with high practicability and validity. Copyright © 2011 Elsevier Ltd. All rights reserved.

  2. Guidelines for the use and interpretation of assays for monitoring autophagy.

    PubMed

    Klionsky, Daniel J; Abdalla, Fabio C; Abeliovich, Hagai; Abraham, Robert T; Acevedo-Arozena, Abraham; Adeli, Khosrow; Agholme, Lotta; Agnello, Maria; Agostinis, Patrizia; Aguirre-Ghiso, Julio A; Ahn, Hyung Jun; Ait-Mohamed, Ouardia; Ait-Si-Ali, Slimane; Akematsu, Takahiko; Akira, Shizuo; Al-Younes, Hesham M; Al-Zeer, Munir A; Albert, Matthew L; Albin, Roger L; Alegre-Abarrategui, Javier; Aleo, Maria Francesca; Alirezaei, Mehrdad; Almasan, Alexandru; Almonte-Becerril, Maylin; Amano, Atsuo; Amaravadi, Ravi; Amarnath, Shoba; Amer, Amal O; Andrieu-Abadie, Nathalie; Anantharam, Vellareddy; Ann, David K; Anoopkumar-Dukie, Shailendra; Aoki, Hiroshi; Apostolova, Nadezda; Arancia, Giuseppe; Aris, John P; Asanuma, Katsuhiko; Asare, Nana Y O; Ashida, Hisashi; Askanas, Valerie; Askew, David S; Auberger, Patrick; Baba, Misuzu; Backues, Steven K; Baehrecke, Eric H; Bahr, Ben A; Bai, Xue-Yuan; Bailly, Yannick; Baiocchi, Robert; Baldini, Giulia; Balduini, Walter; Ballabio, Andrea; Bamber, Bruce A; Bampton, Edward T W; Bánhegyi, Gábor; Bartholomew, Clinton R; Bassham, Diane C; Bast, Robert C; Batoko, Henri; Bay, Boon-Huat; Beau, Isabelle; Béchet, Daniel M; Begley, Thomas J; Behl, Christian; Behrends, Christian; Bekri, Soumeya; Bellaire, Bryan; Bendall, Linda J; Benetti, Luca; Berliocchi, Laura; Bernardi, Henri; Bernassola, Francesca; Besteiro, Sébastien; Bhatia-Kissova, Ingrid; Bi, Xiaoning; Biard-Piechaczyk, Martine; Blum, Janice S; Boise, Lawrence H; Bonaldo, Paolo; Boone, David L; Bornhauser, Beat C; Bortoluci, Karina R; Bossis, Ioannis; Bost, Frédéric; Bourquin, Jean-Pierre; Boya, Patricia; Boyer-Guittaut, Michaël; Bozhkov, Peter V; Brady, Nathan R; Brancolini, Claudio; Brech, Andreas; Brenman, Jay E; Brennand, Ana; Bresnick, Emery H; Brest, Patrick; Bridges, Dave; Bristol, Molly L; Brookes, Paul S; Brown, Eric J; Brumell, John H; Brunetti-Pierri, Nicola; Brunk, Ulf T; Bulman, Dennis E; Bultman, Scott J; Bultynck, Geert; Burbulla, Lena F; Bursch, Wilfried; Butchar, Jonathan P; Buzgariu, Wanda; Bydlowski, Sergio P; Cadwell, Ken; Cahová, Monika; Cai, Dongsheng; Cai, Jiyang; Cai, Qian; Calabretta, Bruno; Calvo-Garrido, Javier; Camougrand, Nadine; Campanella, Michelangelo; Campos-Salinas, Jenny; Candi, Eleonora; Cao, Lizhi; Caplan, Allan B; Carding, Simon R; Cardoso, Sandra M; Carew, Jennifer S; Carlin, Cathleen R; Carmignac, Virginie; Carneiro, Leticia A M; Carra, Serena; Caruso, Rosario A; Casari, Giorgio; Casas, Caty; Castino, Roberta; Cebollero, Eduardo; Cecconi, Francesco; Celli, Jean; Chaachouay, Hassan; Chae, Han-Jung; Chai, Chee-Yin; Chan, David C; Chan, Edmond Y; Chang, Raymond Chuen-Chung; Che, Chi-Ming; Chen, Ching-Chow; Chen, Guang-Chao; Chen, Guo-Qiang; Chen, Min; Chen, Quan; Chen, Steve S-L; Chen, WenLi; Chen, Xi; Chen, Xiangmei; Chen, Xiequn; Chen, Ye-Guang; Chen, Yingyu; Chen, Yongqiang; Chen, Yu-Jen; Chen, Zhixiang; Cheng, Alan; Cheng, Christopher H K; Cheng, Yan; Cheong, Heesun; Cheong, Jae-Ho; Cherry, Sara; Chess-Williams, Russ; Cheung, Zelda H; Chevet, Eric; Chiang, Hui-Ling; Chiarelli, Roberto; Chiba, Tomoki; Chin, Lih-Shen; Chiou, Shih-Hwa; Chisari, Francis V; Cho, Chi Hin; Cho, Dong-Hyung; Choi, Augustine M K; Choi, DooSeok; Choi, Kyeong Sook; Choi, Mary E; Chouaib, Salem; Choubey, Divaker; Choubey, Vinay; Chu, Charleen T; Chuang, Tsung-Hsien; Chueh, Sheau-Huei; Chun, Taehoon; Chwae, Yong-Joon; Chye, Mee-Len; Ciarcia, Roberto; Ciriolo, Maria R; Clague, Michael J; Clark, Robert S B; Clarke, Peter G H; Clarke, Robert; Codogno, Patrice; Coller, Hilary A; Colombo, María I; Comincini, Sergio; Condello, Maria; Condorelli, Fabrizio; Cookson, Mark R; Coombs, Graham H; Coppens, Isabelle; Corbalan, Ramon; Cossart, Pascale; Costelli, Paola; Costes, Safia; Coto-Montes, Ana; Couve, Eduardo; Coxon, Fraser P; Cregg, James M; Crespo, José L; Cronjé, Marianne J; Cuervo, Ana Maria; Cullen, Joseph J; Czaja, Mark J; D'Amelio, Marcello; Darfeuille-Michaud, Arlette; Davids, Lester M; Davies, Faith E; De Felici, Massimo; de Groot, John F; de Haan, Cornelis A M; De Martino, Luisa; De Milito, Angelo; De Tata, Vincenzo; Debnath, Jayanta; Degterev, Alexei; Dehay, Benjamin; Delbridge, Lea M D; Demarchi, Francesca; Deng, Yi Zhen; Dengjel, Jörn; Dent, Paul; Denton, Donna; Deretic, Vojo; Desai, Shyamal D; Devenish, Rodney J; Di Gioacchino, Mario; Di Paolo, Gilbert; Di Pietro, Chiara; Díaz-Araya, Guillermo; Díaz-Laviada, Inés; Diaz-Meco, Maria T; Diaz-Nido, Javier; Dikic, Ivan; Dinesh-Kumar, Savithramma P; Ding, Wen-Xing; Distelhorst, Clark W; Diwan, Abhinav; Djavaheri-Mergny, Mojgan; Dokudovskaya, Svetlana; Dong, Zheng; Dorsey, Frank C; Dosenko, Victor; Dowling, James J; Doxsey, Stephen; Dreux, Marlène; Drew, Mark E; Duan, Qiuhong; Duchosal, Michel A; Duff, Karen; Dugail, Isabelle; Durbeej, Madeleine; Duszenko, Michael; Edelstein, Charles L; Edinger, Aimee L; Egea, Gustavo; Eichinger, Ludwig; Eissa, N Tony; Ekmekcioglu, Suhendan; El-Deiry, Wafik S; Elazar, Zvulun; Elgendy, Mohamed; Ellerby, Lisa M; Eng, Kai Er; Engelbrecht, Anna-Mart; Engelender, Simone; Erenpreisa, Jekaterina; Escalante, Ricardo; Esclatine, Audrey; Eskelinen, Eeva-Liisa; Espert, Lucile; Espina, Virginia; Fan, Huizhou; Fan, Jia; Fan, Qi-Wen; Fan, Zhen; Fang, Shengyun; Fang, Yongqi; Fanto, Manolis; Fanzani, Alessandro; Farkas, Thomas; Farré, Jean-Claude; Faure, Mathias; Fechheimer, Marcus; Feng, Carl G; Feng, Jian; Feng, Qili; Feng, Youji; Fésüs, László; Feuer, Ralph; Figueiredo-Pereira, Maria E; Fimia, Gian Maria; Fingar, Diane C; Finkbeiner, Steven; Finkel, Toren; Finley, Kim D; Fiorito, Filomena; Fisher, Edward A; Fisher, Paul B; Flajolet, Marc; Florez-McClure, Maria L; Florio, Salvatore; Fon, Edward A; Fornai, Francesco; Fortunato, Franco; Fotedar, Rati; Fowler, Daniel H; Fox, Howard S; Franco, Rodrigo; Frankel, Lisa B; Fransen, Marc; Fuentes, José M; Fueyo, Juan; Fujii, Jun; Fujisaki, Kozo; Fujita, Eriko; Fukuda, Mitsunori; Furukawa, Ruth H; Gaestel, Matthias; Gailly, Philippe; Gajewska, Malgorzata; Galliot, Brigitte; Galy, Vincent; Ganesh, Subramaniam; Ganetzky, Barry; Ganley, Ian G; Gao, Fen-Biao; Gao, George F; Gao, Jinming; Garcia, Lorena; Garcia-Manero, Guillermo; Garcia-Marcos, Mikel; Garmyn, Marjan; Gartel, Andrei L; Gatti, Evelina; Gautel, Mathias; Gawriluk, Thomas R; Gegg, Matthew E; Geng, Jiefei; Germain, Marc; Gestwicki, Jason E; Gewirtz, David A; Ghavami, Saeid; Ghosh, Pradipta; Giammarioli, Anna M; Giatromanolaki, Alexandra N; Gibson, Spencer B; Gilkerson, Robert W; Ginger, Michael L; Ginsberg, Henry N; Golab, Jakub; Goligorsky, Michael S; Golstein, Pierre; Gomez-Manzano, Candelaria; Goncu, Ebru; Gongora, Céline; Gonzalez, Claudio D; Gonzalez, Ramon; González-Estévez, Cristina; González-Polo, Rosa Ana; Gonzalez-Rey, Elena; Gorbunov, Nikolai V; Gorski, Sharon; Goruppi, Sandro; Gottlieb, Roberta A; Gozuacik, Devrim; Granato, Giovanna Elvira; Grant, Gary D; Green, Kim N; Gregorc, Aleš; Gros, Frédéric; Grose, Charles; Grunt, Thomas W; Gual, Philippe; Guan, Jun-Lin; Guan, Kun-Liang; Guichard, Sylvie M; Gukovskaya, Anna S; Gukovsky, Ilya; Gunst, Jan; Gustafsson, Asa B; Halayko, Andrew J; Hale, Amber N; Halonen, Sandra K; Hamasaki, Maho; Han, Feng; Han, Ting; Hancock, Michael K; Hansen, Malene; Harada, Hisashi; Harada, Masaru; Hardt, Stefan E; Harper, J Wade; Harris, Adrian L; Harris, James; Harris, Steven D; Hashimoto, Makoto; Haspel, Jeffrey A; Hayashi, Shin-ichiro; Hazelhurst, Lori A; He, Congcong; He, You-Wen; Hébert, Marie-Joseé; Heidenreich, Kim A; Helfrich, Miep H; Helgason, Gudmundur V; Henske, Elizabeth P; Herman, Brian; Herman, Paul K; Hetz, Claudio; Hilfiker, Sabine; Hill, Joseph A; Hocking, Lynne J; Hofman, Paul; Hofmann, Thomas G; Höhfeld, Jörg; Holyoake, Tessa L; Hong, Ming-Huang; Hood, David A; Hotamisligil, Gökhan S; Houwerzijl, Ewout J; Høyer-Hansen, Maria; Hu, Bingren; Hu, Chien-An A; Hu, Hong-Ming; Hua, Ya; Huang, Canhua; Huang, Ju; Huang, Shengbing; Huang, Wei-Pang; Huber, Tobias B; Huh, Won-Ki; Hung, Tai-Ho; Hupp, Ted R; Hur, Gang Min; Hurley, James B; Hussain, Sabah N A; Hussey, Patrick J; Hwang, Jung Jin; Hwang, Seungmin; Ichihara, Atsuhiro; Ilkhanizadeh, Shirin; Inoki, Ken; Into, Takeshi; Iovane, Valentina; Iovanna, Juan L; Ip, Nancy Y; Isaka, Yoshitaka; Ishida, Hiroyuki; Isidoro, Ciro; Isobe, Ken-ichi; Iwasaki, Akiko; Izquierdo, Marta; Izumi, Yotaro; Jaakkola, Panu M; Jäättelä, Marja; Jackson, George R; Jackson, William T; Janji, Bassam; Jendrach, Marina; Jeon, Ju-Hong; Jeung, Eui-Bae; Jiang, Hong; Jiang, Hongchi; Jiang, Jean X; Jiang, Ming; Jiang, Qing; Jiang, Xuejun; Jiang, Xuejun; Jiménez, Alberto; Jin, Meiyan; Jin, Shengkan; Joe, Cheol O; Johansen, Terje; Johnson, Daniel E; Johnson, Gail V W; Jones, Nicola L; Joseph, Bertrand; Joseph, Suresh K; Joubert, Annie M; Juhász, Gábor; Juillerat-Jeanneret, Lucienne; Jung, Chang Hwa; Jung, Yong-Keun; Kaarniranta, Kai; Kaasik, Allen; Kabuta, Tomohiro; Kadowaki, Motoni; Kagedal, Katarina; Kamada, Yoshiaki; Kaminskyy, Vitaliy O; Kampinga, Harm H; Kanamori, Hiromitsu; Kang, Chanhee; Kang, Khong Bee; Kang, Kwang Il; Kang, Rui; Kang, Yoon-A; Kanki, Tomotake; Kanneganti, Thirumala-Devi; Kanno, Haruo; Kanthasamy, Anumantha G; Kanthasamy, Arthi; Karantza, Vassiliki; Kaushal, Gur P; Kaushik, Susmita; Kawazoe, Yoshinori; Ke, Po-Yuan; Kehrl, John H; Kelekar, Ameeta; Kerkhoff, Claus; Kessel, David H; Khalil, Hany; Kiel, Jan A K W; Kiger, Amy A; Kihara, Akio; Kim, Deok Ryong; Kim, Do-Hyung; Kim, Dong-Hou; Kim, Eun-Kyoung; Kim, Hyung-Ryong; Kim, Jae-Sung; Kim, Jeong Hun; Kim, Jin Cheon; Kim, John K; Kim, Peter K; Kim, Seong Who; Kim, Yong-Sun; Kim, Yonghyun; Kimchi, Adi; Kimmelman, Alec C; King, Jason S; Kinsella, Timothy J; Kirkin, Vladimir; Kirshenbaum, Lorrie A; Kitamoto, Katsuhiko; Kitazato, Kaio; Klein, Ludger; Klimecki, Walter T; Klucken, Jochen; Knecht, Erwin; Ko, Ben C B; Koch, Jan C; Koga, Hiroshi; Koh, Jae-Young; Koh, Young Ho; Koike, Masato; Komatsu, Masaaki; Kominami, Eiki; Kong, Hee Jeong; Kong, Wei-Jia; Korolchuk, Viktor I; Kotake, Yaichiro; Koukourakis, Michael I; Kouri Flores, Juan B; Kovács, Attila L; Kraft, Claudine; Krainc, Dimitri; Krämer, Helmut; Kretz-Remy, Carole; Krichevsky, Anna M; Kroemer, Guido; Krüger, Rejko; Krut, Oleg; Ktistakis, Nicholas T; Kuan, Chia-Yi; Kucharczyk, Roza; Kumar, Ashok; Kumar, Raj; Kumar, Sharad; Kundu, Mondira; Kung, Hsing-Jien; Kurz, Tino; Kwon, Ho Jeong; La Spada, Albert R; Lafont, Frank; Lamark, Trond; Landry, Jacques; Lane, Jon D; Lapaquette, Pierre; Laporte, Jocelyn F; László, Lajos; Lavandero, Sergio; Lavoie, Josée N; Layfield, Robert; Lazo, Pedro A; Le, Weidong; Le Cam, Laurent; Ledbetter, Daniel J; Lee, Alvin J X; Lee, Byung-Wan; Lee, Gyun Min; Lee, Jongdae; Lee, Ju-Hyun; Lee, Michael; Lee, Myung-Shik; Lee, Sug Hyung; Leeuwenburgh, Christiaan; Legembre, Patrick; Legouis, Renaud; Lehmann, Michael; Lei, Huan-Yao; Lei, Qun-Ying; Leib, David A; Leiro, José; Lemasters, John J; Lemoine, Antoinette; Lesniak, Maciej S; Lev, Dina; Levenson, Victor V; Levine, Beth; Levy, Efrat; Li, Faqiang; Li, Jun-Lin; Li, Lian; Li, Sheng; Li, Weijie; Li, Xue-Jun; Li, Yan-bo; Li, Yi-Ping; Liang, Chengyu; Liang, Qiangrong; Liao, Yung-Feng; Liberski, Pawel P; Lieberman, Andrew; Lim, Hyunjung J; Lim, Kah-Leong; Lim, Kyu; Lin, Chiou-Feng; Lin, Fu-Cheng; Lin, Jian; Lin, Jiandie D; Lin, Kui; Lin, Wan-Wan; Lin, Weei-Chin; Lin, Yi-Ling; Linden, Rafael; Lingor, Paul; Lippincott-Schwartz, Jennifer; Lisanti, Michael P; Liton, Paloma B; Liu, Bo; Liu, Chun-Feng; Liu, Kaiyu; Liu, Leyuan; Liu, Qiong A; Liu, Wei; Liu, Young-Chau; Liu, Yule; Lockshin, Richard A; Lok, Chun-Nam; Lonial, Sagar; Loos, Benjamin; Lopez-Berestein, Gabriel; López-Otín, Carlos; Lossi, Laura; Lotze, Michael T; Lőw, Peter; Lu, Binfeng; Lu, Bingwei; Lu, Bo; Lu, Zhen; Luciano, Frédéric; Lukacs, Nicholas W; Lund, Anders H; Lynch-Day, Melinda A; Ma, Yong; Macian, Fernando; MacKeigan, Jeff P; Macleod, Kay F; Madeo, Frank; Maiuri, Luigi; Maiuri, Maria Chiara; Malagoli, Davide; Malicdan, May Christine V; Malorni, Walter; Man, Na; Mandelkow, Eva-Maria; Manon, Stéphen; Manov, Irena; Mao, Kai; Mao, Xiang; Mao, Zixu; Marambaud, Philippe; Marazziti, Daniela; Marcel, Yves L; Marchbank, Katie; Marchetti, Piero; Marciniak, Stefan J; Marcondes, Mateus; Mardi, Mohsen; Marfe, Gabriella; Mariño, Guillermo; Markaki, Maria; Marten, Mark R; Martin, Seamus J; Martinand-Mari, Camille; Martinet, Wim; Martinez-Vicente, Marta; Masini, Matilde; Matarrese, Paola; Matsuo, Saburo; Matteoni, Raffaele; Mayer, Andreas; Mazure, Nathalie M; McConkey, David J; McConnell, Melanie J; McDermott, Catherine; McDonald, Christine; McInerney, Gerald M; McKenna, Sharon L; McLaughlin, BethAnn; McLean, Pamela J; McMaster, Christopher R; McQuibban, G Angus; Meijer, Alfred J; Meisler, Miriam H; Meléndez, Alicia; Melia, Thomas J; Melino, Gerry; Mena, Maria A; Menendez, Javier A; Menna-Barreto, Rubem F S; Menon, Manoj B; Menzies, Fiona M; Mercer, Carol A; Merighi, Adalberto; Merry, Diane E; Meschini, Stefania; Meyer, Christian G; Meyer, Thomas F; Miao, Chao-Yu; Miao, Jun-Ying; Michels, Paul A M; Michiels, Carine; Mijaljica, Dalibor; Milojkovic, Ana; Minucci, Saverio; Miracco, Clelia; Miranti, Cindy K; Mitroulis, Ioannis; Miyazawa, Keisuke; Mizushima, Noboru; Mograbi, Baharia; Mohseni, Simin; Molero, Xavier; Mollereau, Bertrand; Mollinedo, Faustino; Momoi, Takashi; Monastyrska, Iryna; Monick, Martha M; Monteiro, Mervyn J; Moore, Michael N; Mora, Rodrigo; Moreau, Kevin; Moreira, Paula I; Moriyasu, Yuji; Moscat, Jorge; Mostowy, Serge; Mottram, Jeremy C; Motyl, Tomasz; Moussa, Charbel E-H; Müller, Sylke; Muller, Sylviane; Münger, Karl; Münz, Christian; Murphy, Leon O; Murphy, Maureen E; Musarò, Antonio; Mysorekar, Indira; Nagata, Eiichiro; Nagata, Kazuhiro; Nahimana, Aimable; Nair, Usha; Nakagawa, Toshiyuki; Nakahira, Kiichi; Nakano, Hiroyasu; Nakatogawa, Hitoshi; Nanjundan, Meera; Naqvi, Naweed I; Narendra, Derek P; Narita, Masashi; Navarro, Miguel; Nawrocki, Steffan T; Nazarko, Taras Y; Nemchenko, Andriy; Netea, Mihai G; Neufeld, Thomas P; Ney, Paul A; Nezis, Ioannis P; Nguyen, Huu Phuc; Nie, Daotai; Nishino, Ichizo; Nislow, Corey; Nixon, Ralph A; Noda, Takeshi; Noegel, Angelika A; Nogalska, Anna; Noguchi, Satoru; Notterpek, Lucia; Novak, Ivana; Nozaki, Tomoyoshi; Nukina, Nobuyuki; Nürnberger, Thorsten; Nyfeler, Beat; Obara, Keisuke; Oberley, Terry D; Oddo, Salvatore; Ogawa, Michinaga; Ohashi, Toya; Okamoto, Koji; Oleinick, Nancy L; Oliver, F Javier; Olsen, Laura J; Olsson, Stefan; Opota, Onya; Osborne, Timothy F; Ostrander, Gary K; Otsu, Kinya; Ou, Jing-hsiung James; Ouimet, Mireille; Overholtzer, Michael; Ozpolat, Bulent; Paganetti, Paolo; Pagnini, Ugo; Pallet, Nicolas; Palmer, Glen E; Palumbo, Camilla; Pan, Tianhong; Panaretakis, Theocharis; Pandey, Udai Bhan; Papackova, Zuzana; Papassideri, Issidora; Paris, Irmgard; Park, Junsoo; Park, Ohkmae K; Parys, Jan B; Parzych, Katherine R; Patschan, Susann; Patterson, Cam; Pattingre, Sophie; Pawelek, John M; Peng, Jianxin; Perlmutter, David H; Perrotta, Ida; Perry, George; Pervaiz, Shazib; Peter, Matthias; Peters, Godefridus J; Petersen, Morten; Petrovski, Goran; Phang, James M; Piacentini, Mauro; Pierre, Philippe; Pierrefite-Carle, Valérie; Pierron, Gérard; Pinkas-Kramarski, Ronit; Piras, Antonio; Piri, Natik; Platanias, Leonidas C; Pöggeler, Stefanie; Poirot, Marc; Poletti, Angelo; Poüs, Christian; Pozuelo-Rubio, Mercedes; Prætorius-Ibba, Mette; Prasad, Anil; Prescott, Mark; Priault, Muriel; Produit-Zengaffinen, Nathalie; Progulske-Fox, Ann; Proikas-Cezanne, Tassula; Przedborski, Serge; Przyklenk, Karin; Puertollano, Rosa; Puyal, Julien; Qian, Shu-Bing; Qin, Liang; Qin, Zheng-Hong; Quaggin, Susan E; Raben, Nina; Rabinowich, Hannah; Rabkin, Simon W; Rahman, Irfan; Rami, Abdelhaq; Ramm, Georg; Randall, Glenn; Randow, Felix; Rao, V Ashutosh; Rathmell, Jeffrey C; Ravikumar, Brinda; Ray, Swapan K; Reed, Bruce H; Reed, John C; Reggiori, Fulvio; Régnier-Vigouroux, Anne; Reichert, Andreas S; Reiners, John J; Reiter, Russel J; Ren, Jun; Revuelta, José L; Rhodes, Christopher J; Ritis, Konstantinos; Rizzo, Elizete; Robbins, Jeffrey; Roberge, Michel; Roca, Hernan; Roccheri, Maria C; Rocchi, Stephane; Rodemann, H Peter; Rodríguez de Córdoba, Santiago; Rohrer, Bärbel; Roninson, Igor B; Rosen, Kirill; Rost-Roszkowska, Magdalena M; Rouis, Mustapha; Rouschop, Kasper M A; Rovetta, Francesca; Rubin, Brian P; Rubinsztein, David C; Ruckdeschel, Klaus; Rucker, Edmund B; Rudich, Assaf; Rudolf, Emil; Ruiz-Opazo, Nelson; Russo, Rossella; Rusten, Tor Erik; Ryan, Kevin M; Ryter, Stefan W; Sabatini, David M; Sadoshima, Junichi; Saha, Tapas; Saitoh, Tatsuya; Sakagami, Hiroshi; Sakai, Yasuyoshi; Salekdeh, Ghasem Hoseini; Salomoni, Paolo; Salvaterra, Paul M; Salvesen, Guy; Salvioli, Rosa; Sanchez, Anthony M J; Sánchez-Alcázar, José A; Sánchez-Prieto, Ricardo; Sandri, Marco; Sankar, Uma; Sansanwal, Poonam; Santambrogio, Laura; Saran, Shweta; Sarkar, Sovan; Sarwal, Minnie; Sasakawa, Chihiro; Sasnauskiene, Ausra; Sass, Miklós; Sato, Ken; Sato, Miyuki; Schapira, Anthony H V; Scharl, Michael; Schätzl, Hermann M; Scheper, Wiep; Schiaffino, Stefano; Schneider, Claudio; Schneider, Marion E; Schneider-Stock, Regine; Schoenlein, Patricia V; Schorderet, Daniel F; Schüller, Christoph; Schwartz, Gary K; Scorrano, Luca; Sealy, Linda; Seglen, Per O; Segura-Aguilar, Juan; Seiliez, Iban; Seleverstov, Oleksandr; Sell, Christian; Seo, Jong Bok; Separovic, Duska; Setaluri, Vijayasaradhi; Setoguchi, Takao; Settembre, Carmine; Shacka, John J; Shanmugam, Mala; Shapiro, Irving M; Shaulian, Eitan; Shaw, Reuben J; Shelhamer, James H; Shen, Han-Ming; Shen, Wei-Chiang; Sheng, Zu-Hang; Shi, Yang; Shibuya, Kenichi; Shidoji, Yoshihiro; Shieh, Jeng-Jer; Shih, Chwen-Ming; Shimada, Yohta; Shimizu, Shigeomi; Shintani, Takahiro; Shirihai, Orian S; Shore, Gordon C; Sibirny, Andriy A; Sidhu, Stan B; Sikorska, Beata; Silva-Zacarin, Elaine C M; Simmons, Alison; Simon, Anna Katharina; Simon, Hans-Uwe; Simone, Cristiano; Simonsen, Anne; Sinclair, David A; Singh, Rajat; Sinha, Debasish; Sinicrope, Frank A; Sirko, Agnieszka; Siu, Parco M; Sivridis, Efthimios; Skop, Vojtech; Skulachev, Vladimir P; Slack, Ruth S; Smaili, Soraya S; Smith, Duncan R; Soengas, Maria S; Soldati, Thierry; Song, Xueqin; Sood, Anil K; Soong, Tuck Wah; Sotgia, Federica; Spector, Stephen A; Spies, Claudia D; Springer, Wolfdieter; Srinivasula, Srinivasa M; Stefanis, Leonidas; Steffan, Joan S; Stendel, Ruediger; Stenmark, Harald; Stephanou, Anastasis; Stern, Stephan T; Sternberg, Cinthya; Stork, Björn; Strålfors, Peter; Subauste, Carlos S; Sui, Xinbing; Sulzer, David; Sun, Jiaren; Sun, Shi-Yong; Sun, Zhi-Jun; Sung, Joseph J Y; Suzuki, Kuninori; Suzuki, Toshihiko; Swanson, Michele S; Swanton, Charles; Sweeney, Sean T; Sy, Lai-King; Szabadkai, Gyorgy; Tabas, Ira; Taegtmeyer, Heinrich; Tafani, Marco; Takács-Vellai, Krisztina; Takano, Yoshitaka; Takegawa, Kaoru; Takemura, Genzou; Takeshita, Fumihiko; Talbot, Nicholas J; Tan, Kevin S W; Tanaka, Keiji; Tanaka, Kozo; Tang, Daolin; Tang, Dingzhong; Tanida, Isei; Tannous, Bakhos A; Tavernarakis, Nektarios; Taylor, Graham S; Taylor, Gregory A; Taylor, J Paul; Terada, Lance S; Terman, Alexei; Tettamanti, Gianluca; Thevissen, Karin; Thompson, Craig B; Thorburn, Andrew; Thumm, Michael; Tian, FengFeng; Tian, Yuan; Tocchini-Valentini, Glauco; Tolkovsky, Aviva M; Tomino, Yasuhiko; Tönges, Lars; Tooze, Sharon A; Tournier, Cathy; Tower, John; Towns, Roberto; Trajkovic, Vladimir; Travassos, Leonardo H; Tsai, Ting-Fen; Tschan, Mario P; Tsubata, Takeshi; Tsung, Allan; Turk, Boris; Turner, Lorianne S; Tyagi, Suresh C; Uchiyama, Yasuo; Ueno, Takashi; Umekawa, Midori; Umemiya-Shirafuji, Rika; Unni, Vivek K; Vaccaro, Maria I; Valente, Enza Maria; Van den Berghe, Greet; van der Klei, Ida J; van Doorn, Wouter; van Dyk, Linda F; van Egmond, Marjolein; van Grunsven, Leo A; Vandenabeele, Peter; Vandenberghe, Wim P; Vanhorebeek, Ilse; Vaquero, Eva C; Velasco, Guillermo; Vellai, Tibor; Vicencio, Jose Miguel; Vierstra, Richard D; Vila, Miquel; Vindis, Cécile; Viola, Giampietro; Viscomi, Maria Teresa; Voitsekhovskaja, Olga V; von Haefen, Clarissa; Votruba, Marcela; Wada, Keiji; Wade-Martins, Richard; Walker, Cheryl L; Walsh, Craig M; Walter, Jochen; Wan, Xiang-Bo; Wang, Aimin; Wang, Chenguang; Wang, Dawei; Wang, Fan; Wang, Fen; Wang, Guanghui; Wang, Haichao; Wang, Hong-Gang; Wang, Horng-Dar; Wang, Jin; Wang, Ke; Wang, Mei; Wang, Richard C; Wang, Xinglong; Wang, Xuejun; Wang, Ying-Jan; Wang, Yipeng; Wang, Zhen; Wang, Zhigang Charles; Wang, Zhinong; Wansink, Derick G; Ward, Diane M; Watada, Hirotaka; Waters, Sarah L; Webster, Paul; Wei, Lixin; Weihl, Conrad C; Weiss, William A; Welford, Scott M; Wen, Long-Ping; Whitehouse, Caroline A; Whitton, J Lindsay; Whitworth, Alexander J; Wileman, Tom; Wiley, John W; Wilkinson, Simon; Willbold, Dieter; Williams, Roger L; Williamson, Peter R; Wouters, Bradly G; Wu, Chenghan; Wu, Dao-Cheng; Wu, William K K; Wyttenbach, Andreas; Xavier, Ramnik J; Xi, Zhijun; Xia, Pu; Xiao, Gengfu; Xie, Zhiping; Xie, Zhonglin; Xu, Da-zhi; Xu, Jianzhen; Xu, Liang; Xu, Xiaolei; Yamamoto, Ai; Yamamoto, Akitsugu; Yamashina, Shunhei; Yamashita, Michiaki; Yan, Xianghua; Yanagida, Mitsuhiro; Yang, Dun-Sheng; Yang, Elizabeth; Yang, Jin-Ming; Yang, Shi Yu; Yang, Wannian; Yang, Wei Yuan; Yang, Zhifen; Yao, Meng-Chao; Yao, Tso-Pang; Yeganeh, Behzad; Yen, Wei-Lien; Yin, Jia-jing; Yin, Xiao-Ming; Yoo, Ook-Joon; Yoon, Gyesoon; Yoon, Seung-Yong; Yorimitsu, Tomohiro; Yoshikawa, Yuko; Yoshimori, Tamotsu; Yoshimoto, Kohki; You, Ho Jin; Youle, Richard J; Younes, Anas; Yu, Li; Yu, Long; Yu, Seong-Woon; Yu, Wai Haung; Yuan, Zhi-Min; Yue, Zhenyu; Yun, Cheol-Heui; Yuzaki, Michisuke; Zabirnyk, Olga; Silva-Zacarin, Elaine; Zacks, David; Zacksenhaus, Eldad; Zaffaroni, Nadia; Zakeri, Zahra; Zeh, Herbert J; Zeitlin, Scott O; Zhang, Hong; Zhang, Hui-Ling; Zhang, Jianhua; Zhang, Jing-Pu; Zhang, Lin; Zhang, Long; Zhang, Ming-Yong; Zhang, Xu Dong; Zhao, Mantong; Zhao, Yi-Fang; Zhao, Ying; Zhao, Zhizhuang J; Zheng, Xiaoxiang; Zhivotovsky, Boris; Zhong, Qing; Zhou, Cong-Zhao; Zhu, Changlian; Zhu, Wei-Guo; Zhu, Xiao-Feng; Zhu, Xiongwei; Zhu, Yuangang; Zoladek, Teresa; Zong, Wei-Xing; Zorzano, Antonio; Zschocke, Jürgen; Zuckerbraun, Brian

    2012-04-01

    In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused

  3. Guidelines for the use and interpretation of assays for monitoring autophagy

    PubMed Central

    Klionsky, Daniel J.; Abdalla, Fabio C.; Abeliovich, Hagai; Abraham, Robert T.; Acevedo-Arozena, Abraham; Adeli, Khosrow; Agholme, Lotta; Agnello, Maria; Agostinis, Patrizia; Aguirre-Ghiso, Julio A.; Ahn, Hyung Jun; Ait-Mohamed, Ouardia; Ait-Si-Ali, Slimane; Akematsu, Takahiko; Akira, Shizuo; Al-Younes, Hesham M.; Al-Zeer, Munir A.; Albert, Matthew L.; Albin, Roger L.; Alegre-Abarrategui, Javier; Aleo, Maria Francesca; Alirezaei, Mehrdad; Almasan, Alexandru; Almonte-Becerril, Maylin; Amano, Atsuo; Amaravadi, Ravi K.; Amarnath, Shoba; Amer, Amal O.; Andrieu-Abadie, Nathalie; Anantharam, Vellareddy; Ann, David K.; Anoopkumar-Dukie, Shailendra; Aoki, Hiroshi; Apostolova, Nadezda; Arancia, Giuseppe; Aris, John P.; Asanuma, Katsuhiko; Asare, Nana Y.O.; Ashida, Hisashi; Askanas, Valerie; Askew, David S.; Auberger, Patrick; Baba, Misuzu; Backues, Steven K.; Baehrecke, Eric H.; Bahr, Ben A.; Bai, Xue-Yuan; Bailly, Yannick; Baiocchi, Robert; Baldini, Giulia; Balduini, Walter; Ballabio, Andrea; Bamber, Bruce A.; Bampton, Edward T.W.; Juhász, Gábor; Bartholomew, Clinton R.; Bassham, Diane C.; Bast, Robert C.; Batoko, Henri; Bay, Boon-Huat; Beau, Isabelle; Béchet, Daniel M.; Begley, Thomas J.; Behl, Christian; Behrends, Christian; Bekri, Soumeya; Bellaire, Bryan; Bendall, Linda J.; Benetti, Luca; Berliocchi, Laura; Bernardi, Henri; Bernassola, Francesca; Besteiro, Sébastien; Bhatia-Kissova, Ingrid; Bi, Xiaoning; Biard-Piechaczyk, Martine; Blum, Janice S.; Boise, Lawrence H.; Bonaldo, Paolo; Boone, David L.; Bornhauser, Beat C.; Bortoluci, Karina R.; Bossis, Ioannis; Bost, Frédéric; Bourquin, Jean-Pierre; Boya, Patricia; Boyer-Guittaut, Michaël; Bozhkov, Peter V.; Brady, Nathan R; Brancolini, Claudio; Brech, Andreas; Brenman, Jay E.; Brennand, Ana; Bresnick, Emery H.; Brest, Patrick; Bridges, Dave; Bristol, Molly L.; Brookes, Paul S.; Brown, Eric J.; Brumell, John H.; Brunetti-Pierri, Nicola; Brunk, Ulf T.; Bulman, Dennis E.; Bultman, Scott J.; Bultynck, Geert; Burbulla, Lena F.; Bursch, Wilfried; Butchar, Jonathan P.; Buzgariu, Wanda; Bydlowski, Sergio P.; Cadwell, Ken; Cahová, Monika; Cai, Dongsheng; Cai, Jiyang; Cai, Qian; Calabretta, Bruno; Calvo-Garrido, Javier; Camougrand, Nadine; Campanella, Michelangelo; Campos-Salinas, Jenny; Candi, Eleonora; Cao, Lizhi; Caplan, Allan B.; Carding, Simon R.; Cardoso, Sandra M.; Carew, Jennifer S.; Carlin, Cathleen R.; Carmignac, Virginie; Carneiro, Leticia A.M.; Carra, Serena; Caruso, Rosario A.; Casari, Giorgio; Casas, Caty; Castino, Roberta; Cebollero, Eduardo; Cecconi, Francesco; Celli, Jean; Chaachouay, Hassan; Chae, Han-Jung; Chai, Chee-Yin; Chan, David C.; Chan, Edmond Y.; Chang, Raymond Chuen-Chung; Che, Chi-Ming; Chen, Ching-Chow; Chen, Guang-Chao; Chen, Guo-Qiang; Chen, Min; Chen, Quan; Chen, Steve S.-L.; Chen, WenLi; Chen, Xi; Chen, Xiangmei; Chen, Xiequn; Chen, Ye-Guang; Chen, Yingyu; Chen, Yongqiang; Chen, Yu-Jen; Chen, Zhixiang; Cheng, Alan; Cheng, Christopher H.K.; Cheng, Yan; Cheong, Heesun; Cheong, Jae-Ho; Cherry, Sara; Chess-Williams, Russ; Cheung, Zelda H.; Chevet, Eric; Chiang, Hui-Ling; Chiarelli, Roberto; Chiba, Tomoki; Chin, Lih-Shen; Chiou, Shih-Hwa; Chisari, Francis V.; Cho, Chi Hin; Cho, Dong-Hyung; Choi, Augustine M.K.; Choi, DooSeok; Choi, Kyeong Sook; Choi, Mary E.; Chouaib, Salem; Choubey, Divaker; Choubey, Vinay; Chu, Charleen T.; Chuang, Tsung-Hsien; Chueh, Sheau-Huei; Chun, Taehoon; Chwae, Yong-Joon; Chye, Mee-Len; Ciarcia, Roberto; Ciriolo, Maria R.; Clague, Michael J.; Clark, Robert S.B.; Clarke, Peter G.H.; Clarke, Robert; Codogno, Patrice; Coller, Hilary A.; Colombo, María I.; Comincini, Sergio; Condello, Maria; Condorelli, Fabrizio; Cookson, Mark R.; Coombs, Graham H.; Coppens, Isabelle; Corbalan, Ramon; Cossart, Pascale; Costelli, Paola; Costes, Safia; Coto-Montes, Ana; Couve, Eduardo; Coxon, Fraser P.; Cregg, James M.; Crespo, José L.; Cronjé, Marianne J.; Cuervo, Ana Maria; Cullen, Joseph J.; Czaja, Mark J.; D'Amelio, Marcello; Darfeuille-Michaud, Arlette; Davids, Lester M.; Davies, Faith E.; De Felici, Massimo; de Groot, John F.; de Haan, Cornelis A.M.; De Martino, Luisa; De Milito, Angelo; De Tata, Vincenzo; Debnath, Jayanta; Degterev, Alexei; Dehay, Benjamin; Delbridge, Lea M.D.; Demarchi, Francesca; Deng, Yi Zhen; Dengjel, Jörn; Dent, Paul; Denton, Donna; Deretic, Vojo; Desai, Shyamal D.; Devenish, Rodney J.; Di Gioacchino, Mario; Di Paolo, Gilbert; Di Pietro, Chiara; Díaz-Araya, Guillermo; Díaz-Laviada, Inés; Diaz-Meco, Maria T.; Diaz-Nido, Javier; Dikic, Ivan; Dinesh-Kumar, Savithramma P.; Ding, Wen-Xing; Distelhorst, Clark W.; Diwan, Abhinav; Djavaheri-Mergny, Mojgan; Dokudovskaya, Svetlana; Dong, Zheng; Dorsey, Frank C.; Dosenko, Victor; Dowling, James J.; Doxsey, Stephen; Dreux, Marlène; Drew, Mark E.; Duan, Qiuhong; Duchosal, Michel A.; Duff, Karen E.; Dugail, Isabelle; Durbeej, Madeleine; Duszenko, Michael; Edelstein, Charles L.; Edinger, Aimee L.; Egea, Gustavo; Eichinger, Ludwig; Eissa, N. Tony; Ekmekcioglu, Suhendan; El-Deiry, Wafik S.; Elazar, Zvulun; Elgendy, Mohamed; Ellerby, Lisa M.; Eng, Kai Er; Engelbrecht, Anna-Mart; Engelender, Simone; Erenpreisa, Jekaterina; Escalante, Ricardo; Esclatine, Audrey; Eskelinen, Eeva-Liisa; Espert, Lucile; Espina, Virginia; Fan, Huizhou; Fan, Jia; Fan, Qi-Wen; Fan, Zhen; Fang, Shengyun; Fang, Yongqi; Fanto, Manolis; Fanzani, Alessandro; Farkas, Thomas; Farre, Jean-Claude; Faure, Mathias; Fechheimer, Marcus; Feng, Carl G.; Feng, Jian; Feng, Qili; Feng, Youji; Fésüs, László; Feuer, Ralph; Figueiredo-Pereira, Maria E.; Fimia, Gian Maria; Fingar, Diane C.; Finkbeiner, Steven; Finkel, Toren; Finley, Kim D.; Fiorito, Filomena; Fisher, Edward A.; Fisher, Paul B.; Flajolet, Marc; Florez-McClure, Maria L.; Florio, Salvatore; Fon, Edward A.; Fornai, Francesco; Fortunato, Franco; Fotedar, Rati; Fowler, Daniel H.; Fox, Howard S.; Franco, Rodrigo; Frankel, Lisa B.; Fransen, Marc; Fuentes, José M.; Fueyo, Juan; Fujii, Jun; Fujisaki, Kozo; Fujita, Eriko; Fukuda, Mitsunori; Furukawa, Ruth H.; Gaestel, Matthias; Gailly, Philippe; Gajewska, Malgorzata; Galliot, Brigitte; Galy, Vincent; Ganesh, Subramaniam; Ganetzky, Barry; Ganley, Ian G.; Gao, Fen-Biao; Gao, George F.; Gao, Jinming; Garcia, Lorena; Garcia-Manero, Guillermo; Garcia-Marcos, Mikel; Garmyn, Marjan; Gartel, Andrei L.; Gatti, Evelina; Gautel, Mathias; Gawriluk, Thomas R.; Gegg, Matthew E.; Geng, Jiefei; Germain, Marc; Gestwicki, Jason E.; Gewirtz, David A.; Ghavami, Saeid; Ghosh, Pradipta; Giammarioli, Anna M.; Giatromanolaki, Alexandra N.; Gibson, Spencer B.; Gilkerson, Robert W.; Ginger, Michael L.; Ginsberg, Henry N.; Golab, Jakub; Goligorsky, Michael S.; Golstein, Pierre; Gomez-Manzano, Candelaria; Goncu, Ebru; Gongora, Céline; Gonzalez, Claudio D.; Gonzalez, Ramon; González-Estévez, Cristina; González-Polo, Rosa Ana; Gonzalez-Rey, Elena; Gorbunov, Nikolai V.; Gorski, Sharon; Goruppi, Sandro; Gottlieb, Roberta A.; Gozuacik, Devrim; Granato, Giovanna Elvira; Grant, Gary D.; Green, Kim N.; Gregorc, Ales; Gros, Frédéric; Grose, Charles; Grunt, Thomas W.; Gual, Philippe; Guan, Jun-Lin; Guan, Kun-Liang; Guichard, Sylvie M.; Gukovskaya, Anna S.; Gukovsky, Ilya; Gunst, Jan; Gustafsson, Åsa B.; Halayko, Andrew J.; Hale, Amber N.; Halonen, Sandra K.; Hamasaki, Maho; Han, Feng; Han, Ting; Hancock, Michael K.; Hansen, Malene; Harada, Hisashi; Harada, Masaru; Hardt, Stefan E.; Harper, J. Wade; Harris, Adrian L.; Harris, James; Harris, Steven D.; Hashimoto, Makoto; Haspel, Jeffrey A.; Hayashi, Shin-ichiro; Hazelhurst, Lori A.; He, Congcong; He, You-Wen; Hébert, Marie-Josée; Heidenreich, Kim A.; Helfrich, Miep H.; Helgason, Gudmundur V.; Henske, Elizabeth P.; Herman, Brian; Herman, Paul K.; Hetz, Claudio; Hilfiker, Sabine; Hill, Joseph A.; Hocking, Lynne J.; Hofman, Paul; Hofmann, Thomas G.; Höhfeld, Jörg; Holyoake, Tessa L.; Hong, Ming-Huang; Hood, David A.; Hotamisligil, Gökhan S.; Houwerzijl, Ewout J.; Høyer-Hansen, Maria; Hu, Bingren; Hu, Chien-an A.; Hu, Hong-Ming; Hua, Ya; Huang, Canhua; Huang, Ju; Huang, Shengbing; Huang, Wei-Pang; Huber, Tobias B.; Huh, Won-Ki; Hung, Tai-Ho; Hupp, Ted R.; Hur, Gang Min; Hurley, James B.; Hussain, Sabah N.A.; Hussey, Patrick J.; Hwang, Jung Jin; Hwang, Seungmin; Ichihara, Atsuhiro; Ilkhanizadeh, Shirin; Inoki, Ken; Into, Takeshi; Iovane, Valentina; Iovanna, Juan L.; Ip, Nancy Y.; Isaka, Yoshitaka; Ishida, Hiroyuki; Isidoro, Ciro; Isobe, Ken-ichi; Iwasaki, Akiko; Izquierdo, Marta; Izumi, Yotaro; Jaakkola, Panu M.; Jäättelä, Marja; Jackson, George R.; Jackson, William T.; Janji, Bassam; Jendrach, Marina; Jeon, Ju-Hong; Jeung, Eui-Bae; Jiang, Hong; Jiang, Hongchi; Jiang, Jean X.; Jiang, Ming; Jiang, Qing; Jiang, Xuejun; Jiang, Xuejun; Jiménez, Alberto; Jin, Meiyan; Jin, Shengkan V.; Joe, Cheol O.; Johansen, Terje; Johnson, Daniel E.; Johnson, Gail V.W.; Jones, Nicola L.; Joseph, Bertrand; Joseph, Suresh K.; Joubert, Annie M.; Juhász, Gábor; Juillerat-Jeanneret, Lucienne; Jung, Chang Hwa; Jung, Yong-Keun; Kaarniranta, Kai; Kaasik, Allen; Kabuta, Tomohiro; Kadowaki, Motoni; Kågedal, Katarina; Kamada, Yoshiaki; Kaminskyy, Vitaliy O.; Kampinga, Harm H.; Kanamori, Hiromitsu; Kang, Chanhee; Kang, Khong Bee; Kang, Kwang Il; Kang, Rui; Kang, Yoon-A; Kanki, Tomotake; Kanneganti, Thirumala-Devi; Kanno, Haruo; Kanthasamy, Anumantha G.; Kanthasamy, Arthi; Karantza, Vassiliki; Kaushal, Gur P.; Kaushik, Susmita; Kawazoe, Yoshinori; Ke, Po-Yuan; Kehrl, John H.; Kelekar, Ameeta; Kerkhoff, Claus; Kessel, David H.; Khalil, Hany; Kiel, Jan A.K.W.; Kiger, Amy A.; Kihara, Akio; Kim, Deok Ryong; Kim, Do-Hyung; Kim, Dong-Hou; Kim, Eun-Kyoung; Kim, Hyung-Ryong; Kim, Jae-Sung; Kim, Jeong Hun; Kim, Jin Cheon; Kim, John K.; Kim, Peter K.; Kim, Seong Who; Kim, Yong-Sun; Kim, Yonghyun; Kimchi, Adi; Kimmelman, Alec C.; King, Jason S.; Kinsella, Timothy J.; Kirkin, Vladimir; Kirshenbaum, Lorrie A.; Kitamoto, Katsuhiko; Kitazato, Kaio; Klein, Ludger; Klimecki, Walter T.; Klucken, Jochen; Knecht, Erwin; Ko, Ben C.B.; Koch, Jan C.; Koga, Hiroshi; Koh, Jae-Young; Koh, Young Ho; Koike, Masato; Komatsu, Masaaki; Kominami, Eiki; Kong, Hee Jeong; Kong, Wei-Jia; Korolchuk, Viktor I.; Kotake, Yaichiro; Koukourakis, Michael I.; Flores, Juan B. Kouri; Kovács, Attila L.; Kraft, Claudine; Krainc, Dimitri; Krämer, Helmut; Kretz-Remy, Carole; Krichevsky, Anna M.; Kroemer, Guido; Krüger, Rejko; Krut, Oleg; Ktistakis, Nicholas T.; Kuan, Chia-Yi; Kucharczyk, Roza; Kumar, Ashok; Kumar, Raj; Kumar, Sharad; Kundu, Mondira; Kung, Hsing-Jien; Kurz, Tino; Kwon, Ho Jeong; La Spada, Albert R.; Lafont, Frank; Lamark, Trond; Landry, Jacques; Lane, Jon D.; Lapaquette, Pierre; Laporte, Jocelyn F.; László, Lajos; Lavandero, Sergio; Lavoie, Josée N.; Layfield, Robert; Lazo, Pedro A.; Le, Weidong; Le Cam, Laurent; Ledbetter, Daniel J.; Lee, Alvin J.X.; Lee, Byung-Wan; Lee, Gyun Min; Lee, Jongdae; lee, Ju-hyun; Lee, Michael; Lee, Myung-Shik; Lee, Sug Hyung; Leeuwenburgh, Christiaan; Legembre, Patrick; Legouis, Renaud; Lehmann, Michael; Lei, Huan-Yao; Lei, Qun-Ying; Leib, David A.; Leiro, José; Lemasters, John J.; Lemoine, Antoinette; Lesniak, Maciej S.; Lev, Dina; Levenson, Victor V.; Levine, Beth; Levy, Efrat; Li, Faqiang; Li, Jun-Lin; Li, Lian; Li, Sheng; Li, Weijie; Li, Xue-Jun; Li, Yan-Bo; Li, Yi-Ping; Liang, Chengyu; Liang, Qiangrong; Liao, Yung-Feng; Liberski, Pawel P.; Lieberman, Andrew; Lim, Hyunjung J.; Lim, Kah-Leong; Lim, Kyu; Lin, Chiou-Feng; Lin, Fu-Cheng; Lin, Jian; Lin, Jiandie D.; Lin, Kui; Lin, Wan-Wan; Lin, Weei-Chin; Lin, Yi-Ling; Linden, Rafael; Lingor, Paul; Lippincott-Schwartz, Jennifer; Lisanti, Michael P.; Liton, Paloma B.; Liu, Bo; Liu, Chun-Feng; Liu, Kaiyu; Liu, Leyuan; Liu, Qiong A.; Liu, Wei; Liu, Young-Chau; Liu, Yule; Lockshin, Richard A.; Lok, Chun-Nam; Lonial, Sagar; Loos, Benjamin; Lopez-Berestein, Gabriel; López-Otín, Carlos; Lossi, Laura; Lotze, Michael T.; Low, Peter; Lu, Binfeng; Lu, Bingwei; Lu, Bo; Lu, Zhen; Luciano, Fréderic; Lukacs, Nicholas W.; Lund, Anders H.; Lynch-Day, Melinda A.; Ma, Yong; Macian, Fernando; MacKeigan, Jeff P.; Macleod, Kay F.; Madeo, Frank; Maiuri, Luigi; Maiuri, Maria Chiara; Malagoli, Davide; Malicdan, May Christine V.; Malorni, Walter; Man, Na; Mandelkow, Eva-Maria; Manon, Stephen; Manov, Irena; Mao, Kai; Mao, Xiang; Mao, Zixu; Marambaud, Philippe; Marazziti, Daniela; Marcel, Yves L.; Marchbank, Katie; Marchetti, Piero; Marciniak, Stefan J.; Marcondes, Mateus; Mardi, Mohsen; Marfe, Gabriella; Mariño, Guillermo; Markaki, Maria; Marten, Mark R.; Martin, Seamus J.; Martinand-Mari, Camille; Martinet, Wim; Martinez-Vicente, Marta; Masini, Matilde; Matarrese, Paola; Matsuo, Saburo; Matteoni, Raffaele; Mayer, Andreas; Mazure, Nathalie M.; McConkey, David J.; McConnell, Melanie J.; McDermott, Catherine; McDonald, Christine; McInerney, Gerald M.; McKenna, Sharon L.; McLaughlin, BethAnn; McLean, Pamela J.; McMaster, Christopher R.; McQuibban, G. Angus; Meijer, Alfred J.; Meisler, Miriam H.; Meléndez, Alicia; Melia, Thomas J.; Melino, Gerry; Mena, Maria A.; Menendez, Javier A.; Menna-Barreto, Rubem F. S.; Menon, Manoj B.; Menzies, Fiona M.; Mercer, Carol A.; Merighi, Adalberto; Merry, Diane E.; Meschini, Stefania; Meyer, Christian G.; Meyer, Thomas F.; Miao, Chao-Yu; Miao, Jun-Ying; Michels, Paul A.M.; Michiels, Carine; Mijaljica, Dalibor; Milojkovic, Ana; Minucci, Saverio; Miracco, Clelia; Miranti, Cindy K.; Mitroulis, Ioannis; Miyazawa, Keisuke; Mizushima, Noboru; Mograbi, Baharia; Mohseni, Simin; Molero, Xavier; Mollereau, Bertrand; Mollinedo, Faustino; Momoi, Takashi; Monastyrska, Iryna; Monick, Martha M.; Monteiro, Mervyn J.; Moore, Michael N.; Mora, Rodrigo; Moreau, Kevin; Moreira, Paula I.; Moriyasu, Yuji; Moscat, Jorge; Mostowy, Serge; Mottram, Jeremy C.; Motyl, Tomasz; Moussa, Charbel E.-H.; Müller, Sylke; Muller, Sylviane; Münger, Karl; Münz, Christian; Murphy, Leon O.; Murphy, Maureen E.; Musarò, Antonio; Mysorekar, Indira; Nagata, Eiichiro; Nagata, Kazuhiro; Nahimana, Aimable; Nair, Usha; Nakagawa, Toshiyuki; Nakahira, Kiichi; Nakano, Hiroyasu; Nakatogawa, Hitoshi; Nanjundan, Meera; Naqvi, Naweed I.; Narendra, Derek P.; Narita, Masashi; Navarro, Miguel; Nawrocki, Steffan T.; Nazarko, Taras Y.; Nemchenko, Andriy; Netea, Mihai G.; Neufeld, Thomas P.; Ney, Paul A.; Nezis, Ioannis P.; Nguyen, Huu Phuc; Nie, Daotai; Nishino, Ichizo; Nislow, Corey; Nixon, Ralph A.; Noda, Takeshi; Noegel, Angelika A.; Nogalska, Anna; Noguchi, Satoru; Notterpek, Lucia; Novak, Ivana; Nozaki, Tomoyoshi; Nukina, Nobuyuki; Nürnberger, Thorsten; Nyfeler, Beat; Obara, Keisuke; Oberley, Terry D.; Oddo, Salvatore; Ogawa, Michinaga; Ohashi, Toya; Okamoto, Koji; Oleinick, Nancy L.; Oliver, F. Javier; Olsen, Laura J.; Olsson, Stefan; Opota, Onya; Osborne, Timothy F.; Ostrander, Gary K.; Otsu, Kinya; Ou, Jing-hsiung James; Ouimet, Mireille; Overholtzer, Michael; Ozpolat, Bulent; Paganetti, Paolo; Pagnini, Ugo; Pallet, Nicolas; Palmer, Glen E.; Palumbo, Camilla; Pan, Tianhong; Panaretakis, Theocharis; Pandey, Udai Bhan; Papackova, Zuzana; Papassideri, Issidora; Paris, Irmgard; Park, Junsoo; Park, Ohkmae K.; Parys, Jan B.; Parzych, Katherine R.; Patschan, Susann; Patterson, Cam; Pattingre, Sophie; Pawelek, John M.; Peng, Jianxin; Perlmutter, David H.; Perrotta, Ida; Perry, George; Pervaiz, Shazib; Peter, Matthias; Peters, Godefridus J.; Petersen, Morten; Petrovski, Goran; Phang, James M.; Piacentini, Mauro; Pierre, Philippe; Pierrefite-Carle, Valérie; Pierron, Gérard; Pinkas-Kramarski, Ronit; Piras, Antonio; Piri, Natik; Platanias, Leonidas C.; Pöggeler, Stefanie; Poirot, Marc; Poletti, Angelo; Poüs, Christian; Pozuelo-Rubio, Mercedes; Prætorius-Ibba, Mette; Prasad, Anil; Prescott, Mark; Priault, Muriel; Produit-Zengaffinen, Nathalie; Progulske-Fox, Ann; Proikas-Cezanne, Tassula; Przedborski, Serge; Przyklenk, Karin; Puertollano, Rosa; Puyal, Julien; Qian, Shu-Bing; Qin, Liang; Qin, Zheng-Hong; Quaggin, Susan E.; Raben, Nina; Rabinowich, Hannah; Rabkin, Simon W.; Rahman, Irfan; Rami, Abdelhaq; Ramm, Georg; Randall, Glenn; Randow, Felix; Rao, V. Ashutosh; Rathmell, Jeffrey C.; Ravikumar, Brinda; Ray, Swapan K.; Reed, Bruce H.; Reed, John C.; Reggiori, Fulvio; Régnier-Vigouroux, Anne; Reichert, Andreas S.; Reiners, John J.; Reiter, Russel J.; Ren, Jun; Revuelta, José L.; Rhodes, Christopher J.; Ritis, Konstantinos; Rizzo, Elizete; Robbins, Jeffrey; Roberge, Michel; Roca, Hernan; Roccheri, Maria C.; Rocchi, Stephane; Rodemann, H. Peter; Rodríguez de Córdoba, Santiago; Rohrer, Bärbel; Roninson, Igor B.; Rosen, Kirill; Rost-Roszkowska, Magdalena M.; Rouis, Mustapha; Rouschop, Kasper M.A.; Rovetta, Francesca; Rubin, Brian P.; Rubinsztein, David C.; Ruckdeschel, Klaus; Rucker, Edmund B.; Rudich, Assaf; Rudolf, Emil; Ruiz-Opazo, Nelson; Russo, Rossella; Rusten, Tor Erik; Ryan, Kevin M.; Ryter, Stefan W.; Sabatini, David M.; Sadoshima, Junichi; Saha, Tapas; Saitoh, Tatsuya; Sakagami, Hiroshi; Sakai, Yasuyoshi; Salekdeh, Ghasem Hoseini; Salomoni, Paolo; Salvaterra, Paul M.; Salvesen, Guy; Salvioli, Rosa; Sanchez, Anthony M.J.; Sánchez-Alcázar, José A.; Sánchez-Prieto, Ricardo; Sandri, Marco; Sankar, Uma; Sansanwal, Poonam; Santambrogio, Laura; Saran, Shweta; Sarkar, Sovan; Sarwal, Minnie; Sasakawa, Chihiro; Sasnauskiene, Ausra; Sass, Miklós; Sato, Ken; Sato, Miyuki; Schapira, Anthony H.V.; Scharl, Michael; Schätzl, Hermann M.; Scheper, Wiep; Schiaffino, Stefano; Schneider, Claudio; Schneider, Marion E.; Schneider-Stock, Regine; Schoenlein, Patricia V.; Schorderet, Daniel F.; Schüller, Christoph; Schwartz, Gary K.; Scorrano, Luca; Sealy, Linda; Seglen, Per O.; Segura-Aguilar, Juan; Seiliez, Iban; Seleverstov, Oleksandr; Sell, Christian; Seo, Jong Bok; Separovic, Duska; Setaluri, Vijayasaradhi; Setoguchi, Takao; Settembre, Carmine; Shacka, John J.; Shanmugam, Mala; Shapiro, Irving M.; Shaulian, Eitan; Shaw, Reuben J.; Shelhamer, James H.; Shen, Han-Ming; Shen, Wei-Chiang; Sheng, Zu-Hang; Shi, Yang; Shibuya, Kenichi; Shidoji, Yoshihiro; Shieh, Jeng-Jer; Shih, Chwen-Ming; Shimada, Yohta; Shimizu, Shigeomi; Shintani, Takahiro; Shirihai, Orian S.; Shore, Gordon C.; Sibirny, Andriy A.; Sidhu, Stan B.; Sikorska, Beata; Silva-Zacarin, Elaine C.M.; Simmons, Alison; Simon, Anna Katharina; Simon, Hans-Uwe; Simone, Cristiano; Simonsen, Anne; Sinclair, David A.; Singh, Rajat; Sinha, Debasish; Sinicrope, Frank A.; Sirko, Agnieszka; Siu, Parco M.; Sivridis, Efthimios; Skop, Vojtech; Skulachev, Vladimir P.; Slack, Ruth S.; Smaili, Soraya S.; Smith, Duncan R.; Soengas, Maria S.; Soldati, Thierry; Song, Xueqin; Sood, Anil K.; Soong, Tuck Wah; Sotgia, Federica; Spector, Stephen A.; Spies, Claudia D.; Springer, Wolfdieter; Srinivasula, Srinivasa M.; Stefanis, Leonidas; Steffan, Joan S.; Stendel, Ruediger; Stenmark, Harald; Stephanou, Anastasis; Stern, Stephan T.; Sternberg, Cinthya; Stork, Björn; Strålfors, Peter; Subauste, Carlos S.; Sui, Xinbing; Sulzer, David; Sun, Jiaren; Sun, Shi-Yong; Sun, Zhi-Jun; Sung, Joseph J.Y.; Suzuki, Kuninori; Suzuki, Toshihiko; Swanson, Michele S.; Swanton, Charles; Sweeney, Sean T.; Sy, Lai-King; Szabadkai, György; Tabas, Ira; Taegtmeyer, Heinrich; Tafani, Marco; Takács-Vellai, Krisztina; Takano, Yoshitaka; Takegawa, Kaoru; Takemura, Genzou; Takeshita, Fumihiko; Talbot, Nicholas J.; Tan, Kevin S.W.; Tanaka, Keiji; Tanaka, Kozo; Tang, Daolin; Tang, Dingzhong; Tanida, Isei; Tannous, Bakhos A.; Tavernarakis, Nektarios; Taylor, Graham S.; Taylor, Gregory A.; Taylor, J. Paul; Terada, Lance S.; Terman, Alexei; Tettamanti, Gianluca; Thevissen, Karin; Thompson, Craig B.; Thorburn, Andrew; Thumm, Michael; Tian, FengFeng; Tian, Yuan; Tocchini-Valentini, Glauco; Tolkovsky, Aviva M.; Tomino, Yasuhiko; Tönges, Lars; Tooze, Sharon A.; Tournier, Cathy; Tower, John; Towns, Roberto; Trajkovic, Vladimir; Travassos, Leonardo H.; Tsai, Ting-Fen; Tschan, Mario P.; Tsubata, Takeshi; Tsung, Allan; Turk, Boris; Turner, Lorianne S.; Tyagi, Suresh C.; Uchiyama, Yasuo; Ueno, Takashi; Umekawa, Midori; Umemiya-Shirafuji, Rika; Unni, Vivek K.; Vaccaro, Maria I.; Valente, Enza Maria; Van den Berghe, Greet; van der Klei, Ida J.; van Doorn, Wouter G.; van Dyk, Linda F.; van Egmond, Marjolein; van Grunsven, Leo A.; Vandenabeele, Peter; Vandenberghe, Wim P.; Vanhorebeek, Ilse; Vaquero, Eva C.; Velasco, Guillermo; Vellai, Tibor; Vicencio, José Miguel; Vierstra, Richard D.; Vila, Miquel; Vindis, Cécile; Viola, Giampietro; Viscomi, Maria Teresa; Voitsekhovskaja, Olga V.; von Haefen, Clarissa; Votruba, Marcela; Wada, Keiji; Wade-Martins, Richard; Walker, Cheryl L.; Walsh, Craig M.; Walter, Jochen; Wan, Xiang-Bo; Wang, Aimin; Wang, Chenguang; Wang, Dawei; Wang, Fan; Wang, Fen; Wang, Guanghui; Wang, Haichao; Wang, Hong-Gang; Wang, Horng-Dar; Wang, Jin; Wang, Ke; Wang, Mei; Wang, Richard C.; Wang, Xinglong; Wang, Xiujie J.; Wang, Ying-Jan; Wang, Yipeng; Wang, Zhen-Bo; Wang, Zhigang Charles; Wang, Zhinong; Wansink, Derick G.; Ward, Diane M.; Watada, Hirotaka; Waters, Sarah L.; Webster, Paul; Wei, Lixin; Weihl, Conrad C.; Weiss, William A.; Welford, Scott M.; Wen, Long-Ping; Whitehouse, Caroline A.; Whitton, J. Lindsay; Whitworth, Alexander J.; Wileman, Tom; Wiley, John W.; Wilkinson, Simon; Willbold, Dieter; Williams, Roger L.; Williamson, Peter R.; Wouters, Bradly G.; Wu, Chenghan; Wu, Dao-Cheng; Wu, William K.K.; Wyttenbach, Andreas; Xavier, Ramnik J.; Xi, Zhijun; Xia, Pu; Xiao, Gengfu; Xie, Zhiping; Xie, Zhonglin; Xu, Da-zhi; Xu, Jianzhen; Xu, Liang; Xu, Xiaolei; Yamamoto, Ai; Yamamoto, Akitsugu; Yamashina, Shunhei; Yamashita, Michiaki; Yan, Xianghua; Yanagida, Mitsuhiro; Yang, Dun-Sheng; Yang, Elizabeth; Yang, Jin-Ming; Yang, Shi Yu; Yang, Wannian; Yang, Wei Yuan; Yang, Zhifen; Yao, Meng-Chao; Yao, Tso-Pang; Yeganeh, Behzad; Yen, Wei-Lien; Yin, Jia-Jing; Yin, Xiao-Ming; Yoo, Ook-Joon; Yoon, Gyesoon; Yoon, Seung-Yong; Yorimitsu, Tomohiro; Yoshikawa, Yuko; Yoshimori, Tamotsu; Yoshimoto, Kohki; You, Ho Jin; Youle, Richard J.; Younes, Anas; Yu, Li; Yu, Long; Yu, Seong-Woon; Yu, Wai Haung; Yuan, Zhi-Min; Yue, Zhenyu; Yun, Cheol-Heui; Yuzaki, Michisuke; Zabirnyk, Olga; Silva-Zacarin, Elaine; Zacks, David; Zacksenhaus, Eldad; Zaffaroni, Nadia; Zakeri, Zahra; Zeh, III, Herbert J.; Zeitlin, Scott O.; Zhang, Hong; Zhang, Hui-Ling; Zhang, Jianhua; Zhang, Jing-Pu; Zhang, Lin; Zhang, Long; Zhang, Ming-Yong; Zhang, Xu Dong; Zhao, Mantong; Zhao, Yi-Fang; Zhao, Ying; Zhao, Zhizhuang J.; Zheng, Xiaoxiang; Zhivotovsky, Boris; Zhong, Qing; Zhou, Cong-Zhao; Zhu, Changlian; Zhu, Wei-Guo; Zhu, Xiao-Feng; Zhu, Xiongwei; Zhu, Yuangang; Zoladek, Teresa; Zong, Wei-Xing; Zorzano, Antonio; Zschocke, Jürgen; Zuckerbraun, Brian

    2012-01-01

    In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused

  4. Versatile High-Throughput Fluorescence Assay for Monitoring Cas9 Activity.

    PubMed

    Seamon, Kyle J; Light, Yooli K; Saada, Edwin A; Schoeniger, Joseph S; Harmon, Brooke

    2018-06-05

    The RNA-guided DNA nuclease Cas9 is now widely used for the targeted modification of genomes of human cells and various organisms. Despite the extensive use of Clustered Regularly Interspaced Palindromic Repeats (CRISPR) systems for genome engineering and the rapid discovery and engineering of new CRISPR-associated nucleases, there are no high-throughput assays for measuring enzymatic activity. The current laboratory and future therapeutic uses of CRISPR technology have a significant risk of accidental exposure or clinical off-target effects, underscoring the need for therapeutically effective inhibitors of Cas9. Here, we develop a fluorescence assay for monitoring Cas9 nuclease activity and demonstrate its utility with S. pyogenes (Spy), S. aureus (Sau), and C. jejuni (Cje) Cas9. The assay was validated by quantitatively profiling the species specificity of published anti-CRISPR (Acr) proteins, confirming the reported inhibition of Spy Cas9 by AcrIIA4 and Cje Cas9 by AcrIIC1 and no inhibition of Sau Cas9 by either anti-CRISPR. To identify drug-like inhibitors, we performed a screen of 189 606 small molecules for inhibition of Spy Cas9. Of 437 hits (0.2% hit rate), six were confirmed as Cas9 inhibitors in a direct gel electrophoresis secondary assay. The high-throughput nature of this assay makes it broadly applicable for the discovery of additional Cas9 inhibitors or the characterization of Cas9 enzyme variants.

  5. Versatile High-Throughput Fluorescence Assay for Monitoring Cas9 Activity

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Seamon, Kyle Jeffrey; Light, Yooli Kim; Saada, Edwin A.

    Here, the RNA-guided DNA nuclease Cas9 is now widely used for the targeted modification of genomes of human cells and various organisms. Despite the extensive use of Clustered Regularly Interspaced Palindromic Repeats (CRISPR) systems for genome engineering and the rapid discovery and engineering of new CRISPR-associated nucleases, there are no high-throughput assays for measuring enzymatic activity. The current laboratory and future therapeutic uses of CRISPR technology have a significant risk of accidental exposure or clinical off-target effects, underscoring the need for therapeutically effective inhibitors of Cas9. Here, we develop a fluorescence assay for monitoring Cas9 nuclease activity and demonstrate itsmore » utility with S. pyogenes (Spy), S. aureus (Sau), and C. jejuni (Cje) Cas9. The assay was validated by quantitatively profiling the species specificity of published anti-CRISPR (Acr) proteins, confirming the reported inhibition of Spy Cas9 by AcrIIA4 and Cje Cas9 by AcrIIC1 and no inhibition of Sau Cas9 by either anti-CRISPR. To identify drug-like inhibitors, we performed a screen of 189 606 small molecules for inhibition of Spy Cas9. Of 437 hits (0.2% hit rate), six were confirmed as Cas9 inhibitors in a direct gel electrophoresis secondary assay. The high-throughput nature of this assay makes it broadly applicable for the discovery of additional Cas9 inhibitors or the characterization of Cas9 enzyme variants.« less

  6. Versatile High-Throughput Fluorescence Assay for Monitoring Cas9 Activity

    DOE PAGES

    Seamon, Kyle Jeffrey; Light, Yooli Kim; Saada, Edwin A.; ...

    2018-05-14

    Here, the RNA-guided DNA nuclease Cas9 is now widely used for the targeted modification of genomes of human cells and various organisms. Despite the extensive use of Clustered Regularly Interspaced Palindromic Repeats (CRISPR) systems for genome engineering and the rapid discovery and engineering of new CRISPR-associated nucleases, there are no high-throughput assays for measuring enzymatic activity. The current laboratory and future therapeutic uses of CRISPR technology have a significant risk of accidental exposure or clinical off-target effects, underscoring the need for therapeutically effective inhibitors of Cas9. Here, we develop a fluorescence assay for monitoring Cas9 nuclease activity and demonstrate itsmore » utility with S. pyogenes (Spy), S. aureus (Sau), and C. jejuni (Cje) Cas9. The assay was validated by quantitatively profiling the species specificity of published anti-CRISPR (Acr) proteins, confirming the reported inhibition of Spy Cas9 by AcrIIA4 and Cje Cas9 by AcrIIC1 and no inhibition of Sau Cas9 by either anti-CRISPR. To identify drug-like inhibitors, we performed a screen of 189 606 small molecules for inhibition of Spy Cas9. Of 437 hits (0.2% hit rate), six were confirmed as Cas9 inhibitors in a direct gel electrophoresis secondary assay. The high-throughput nature of this assay makes it broadly applicable for the discovery of additional Cas9 inhibitors or the characterization of Cas9 enzyme variants.« less

  7. Monitoring virologic responses to antiretroviral therapy in HIV-infected adults in Kenya: evaluation of a low-cost viral load assay.

    PubMed

    Sivapalasingam, Sumathi; Wangechi, Beatrice; Marshed, Fatuma; Laverty, Maura; Essajee, Shaffiq; Holzman, Robert S; Valentine, Fred

    2009-08-28

    A key advantage of monitoring HIV viral load (VL) in persons receiving antiretroviral therapy (ART) is the ability to detect virologic failure before clinical deterioration or resistance occurs. Detection of virologic failure will help clarify the need for enhanced adherence counseling or a change to second- line therapy. Low-cost, locally performable alternates to expensive VL assays are needed where resources are limited. We monitored the response to 48-week ART in 100 treatment-naïve Kenyan adults using a low-cost VL measurement, the Cavidi reverse transcriptase (RT) assay and gold-standard assays, Roche RNA PCR and Bayer Versant HIV-1 RNA (bDNA) assays. In Altman-Bland plots, the mean difference in viral loads between the three assays was small (<0.5 log(10) copies/mL). However, the limits of agreement between the methods exceeded the biologically relevant change of 0.5 log copies/ml. Therefore, the RT assay cannot be used interchangeably with the other assays to monitor individual patients. The RT assay was 100% sensitive in detecting viral loads of > or =400 copies/ml compared to gold-standard assays. After 24 weeks of treatment, viral load measured by the RT assay was undetectable in 95% of 65 patients with undetectable RNA PCR VL (<400 copies/ml), 90% of 67 patients with undetectable bDNA VL, and 96% of 57 patients with undetectable VL in both RNA PCR and bDNA assays. The negative predictive value of the RT assay was 100% compared to either assay; the positive predictive value was 86% compared to RNA PCR and 70% compared to bDNA. The RT assay compared well with gold standard assays. Our study highlights the importance of not interchanging viral load assays when monitoring an individual patient. Furthermore, the RT assay may be limited by low positive predictive values when used in populations with low prevalence of virologic failure.

  8. Monitoring Virologic Responses to Antiretroviral Therapy in HIV-Infected Adults in Kenya: Evaluation of a Low-Cost Viral Load Assay

    PubMed Central

    Sivapalasingam, Sumathi; Wangechi, Beatrice; Marshed, Fatuma; Laverty, Maura; Essajee, Shaffiq; Holzman, Robert S.; Valentine, Fred

    2009-01-01

    Background A key advantage of monitoring HIV viral load (VL) in persons receiving antiretroviral therapy (ART) is the ability to detect virologic failure before clinical deterioration or resistance occurs. Detection of virologic failure will help clarify the need for enhanced adherence counseling or a change to second- line therapy. Low-cost, locally performable alternates to expensive VL assays are needed where resources are limited. Methodology/Principal Findings We monitored the response to 48-week ART in 100 treatment-naïve Kenyan adults using a low-cost VL measurement, the Cavidi reverse transcriptase (RT) assay and gold-standard assays, Roche RNA PCR and Bayer Versant HIV-1 RNA (bDNA) assays. In Altman-Bland plots, the mean difference in viral loads between the three assays was small (<0.5 log10 copies/mL). However, the limits of agreement between the methods exceeded the biologically relevant change of 0.5 log copies/ml. Therefore, the RT assay cannot be used interchangeably with the other assays to monitor individual patients. The RT assay was 100% sensitive in detecting viral loads of ≥400 copies/ml compared to gold-standard assays. After 24 weeks of treatment, viral load measured by the RT assay was undetectable in 95% of 65 patients with undetectable RNA PCR VL (<400 copies/ml), 90% of 67 patients with undetectable bDNA VL, and 96% of 57 patients with undetectable VL in both RNA PCR and bDNA assays. The negative predictive value of the RT assay was 100% compared to either assay; the positive predictive value was 86% compared to RNA PCR and 70% compared to bDNA. Conclusion The RT assay compared well with gold standard assays. Our study highlights the importance of not interchanging viral load assays when monitoring an individual patient. Furthermore, the RT assay may be limited by low positive predictive values when used in populations with low prevalence of virologic failure. PMID:19714253

  9. APNEA/WIT system nondestructive assay capability evaluation plan for select accessibly stored INEL RWMC waste forms

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Becker, G.K.

    1997-01-01

    Bio-Imaging Research Inc. (BIR) and Lockheed Martin Speciality Components (LMSC) are engaged in a Program Research and Development Agreement and a Rapid Commercialization Initiative with the Department of Energy, EM-50. The agreement required BIR and LMSC to develop a data interpretation method that merges nondestructive assay and nondestructive examination (NDA/NDE) data and information sufficient to establish compliance with applicable National TRU Program (Program) waste characterization requirements and associated quality assurance performance criteria. This effort required an objective demonstration of the BIR and LMSC waste characterization systems in their standalone and integrated configurations. The goal of the test plan is tomore » provide a mechanism from which evidence can be derived to substantiate nondestructive assay capability and utility statement for the BIT and LMSC systems. The plan must provide for the acquisition, compilation, and reporting of performance data thereby allowing external independent agencies a basis for an objective evaluation of the standalone BIR and LMSC measurement systems, WIT and APNEA respectively, as well as an expected performance resulting from appropriate integration of the two systems. The evaluation is to be structured such that a statement regarding select INEL RWMC waste forms can be made in terms of compliance with applicable Program requirements and criteria.« less

  10. 2016 Waste Management Monitoring Report Area 3 and Area 5 Radioactive Waste Management Sites

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Black, David

    Environmental monitoring data are collected at and around the Area 3 and Area 5 Radioactive Waste Management Sites (RWMSs) within the Nevada National Security Site (NNSS). These data include direct radiation exposure, as well as radiation from the air, groundwater, meteorology, and vadose zone. This report summarizes the 2016 environmental data to provide an overall evaluation of RWMS performance and to support environmental compliance and performance assessment (PA) activities. Some of these data (e.g., radiation exposure, air, and groundwater) are presented in other reports, developed by National Security Technologies, LLC Direct radiation monitoring data indicate exposure levels at the RWMSsmore » are within the range of background levels measured at the NNSS. Slightly elevated exposure levels outside the Area 3 RWMS are attributed to nearby historical aboveground nuclear weapons tests. Air monitoring data show that tritium concentrations in water vapor and americium and plutonium concentrations in air particles are below Derived Concentration Standards for these radionuclides. Groundwater monitoring data indicate the groundwater in the uppermost aquifer beneath the Area 5 RWMS is not impacted by RWMS operations. Results of groundwater analysis from wells around the Area 5 RWMS were all below established investigation levels. Leachate samples collected from the leachate collection system at the mixed low-level waste cell were below established contaminant regulatory limits. During 2016, precipitation at the Area 3 RWMS was 8% below average, and precipitation at the Area 5 RWMS was 8% above average. Water balance measurements indicate that evapotranspiration from the vegetated weighing lysimeter dries the soil and prevents downward percolation of precipitation more effectively than evaporation as measured from the bare-soil weighing lysimeter. Vadose zone monitoring on Area 5 and Area 3 RWMS cell covers shows no evidence of precipitation percolating through the

  11. Separating and stabilizing phosphate from high-level radioactive waste: process development and spectroscopic monitoring.

    PubMed

    Lumetta, Gregg J; Braley, Jenifer C; Peterson, James M; Bryan, Samuel A; Levitskaia, Tatiana G

    2012-06-05

    Removing phosphate from alkaline high-level waste sludges at the Department of Energy's Hanford Site in Washington State is necessary to increase the waste loading in the borosilicate glass waste form that will be used to immobilize the highly radioactive fraction of these wastes. We are developing a process which first leaches phosphate from the high-level waste solids with aqueous sodium hydroxide, and then isolates the phosphate by precipitation with calcium oxide. Tests with actual tank waste confirmed that this process is an effective method of phosphate removal from the sludge and offers an additional option for managing the phosphorus in the Hanford tank waste solids. The presence of vibrationally active species, such as nitrate and phosphate ions, in the tank waste processing streams makes the phosphate removal process an ideal candidate for monitoring by Raman or infrared spectroscopic means. As a proof-of-principle demonstration, Raman and Fourier transform infrared (FTIR) spectra were acquired for all phases during a test of the process with actual tank waste. Quantitative determination of phosphate, nitrate, and sulfate in the liquid phases was achieved by Raman spectroscopy, demonstrating the applicability of Raman spectroscopy for the monitoring of these species in the tank waste process streams.

  12. Passive Neutron Non-Destructive Assay for Remediation of Radiological Waste at Hanford Burial Grounds- 13189

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Simpson, A.; Pitts, M.; Ludowise, J.D.

    The Hanford burial grounds contains a broad spectrum of low activity radioactive wastes, transuranic (TRU) wastes, and hazardous wastes including fission products, byproduct material (thorium and uranium), plutonium and laboratory chemicals. A passive neutron non-destructive assay technique has been developed for characterization of shielded concreted drums exhumed from the burial grounds. This method facilitates the separation of low activity radiological waste containers from TRU waste containers exhumed from the burial grounds. Two identical total neutron counting systems have been deployed, each consisting of He-3 detectors surrounded by a polyethylene moderator. The counts are processed through a statistical filter that removesmore » outliers in order to suppress cosmic spallation events and electronic noise. Upon completion of processing, a 'GO / NO GO' signal is provided to the operator based on a threshold level equivalent to 0.5 grams of weapons grade plutonium in the container being evaluated. This approach allows instantaneous decisions to be made on how to proceed with the waste. The counting systems have been set up using initial on-site measurements (neutron emitting standards loaded into surrogate waste containers) combined with Monte Carlo modeling techniques. The benefit of this approach is to allow the systems to extend their measurement ranges, in terms of applicable matrix types and container sizes, with minimal interruption to the operations at the burial grounds. (authors)« less

  13. Innovations in the Assay of Un-Segregated Multi-Isotopic Grade TRU Waste Boxes with SuperHENC and FRAM Technology

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Simpson, A. P.; Barber, S.; Abdurrahman, N. M.

    2006-07-01

    The Super High Efficiency Neutron Coincidence Counter (SuperHENC) was originally developed by BIL Solutions Inc., Los Alamos National Laboratory (LANL) and Rocky Flats Environmental Technology Site (RFETS) for assay of transuranic (TRU) waste in Standard Waste Boxes (SWB) at Rocky Flats. This mobile system was a key component in the shipment of over 4,000 SWBs to the Waste Isolation Pilot Plant (WIPP) in Carlsbad, New Mexico. The system was WIPP certified in 2001 and operated at the site for four years. The success of this system, a passive neutron coincidence counter combined with high resolution gamma spectroscopy, led to themore » order of two new units, delivered to Hanford in 2004. Several new challenges were faced at Hanford: For example, the original RFETS system was calibrated for segregated waste streams such that metals, plastics, wet combustibles and dry combustibles were separated by 'Item Description Codes' prior to assay. Furthermore, the RFETS mission of handling only weapons grade plutonium, enabled the original SuperHENC to benefit from the use of known Pu isotopics. Operations at Hanford, as with most other DOE sites, generate un-segregated waste streams, with a wide diversity of Pu isotopics. Consequently, the new SuperHENCs are required to deal with new technical challenges. The neutron system's software and calibration methodology have been modified to encompass these new requirements. In addition, PC-FRAM software has been added to the gamma system, providing a robust isotopic measurement capability. Finally a new software package has been developed that integrates the neutron and gamma data to provide a final assay results and analysis report. The new system's performance has been rigorously tested and validated against WIPP quality requirements. These modifications, together with the mobile platform, make the new SuperHENC far more versatile in handling diverse waste streams and allow for rapid redeployment around the DOE complex. (authors)« less

  14. Testing and Performance Validation of a Shielded Waste Segregation and Clearance Monitor Designed for the Measurement of Low Level Waste-13043

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mason, John A.; Burke, Kevin J.; Towner, Antony C.N.

    This paper describes the development, testing and validation of a shielded waste segregation and clearance monitor designed for the measurement of low-density low-level waste (LLW). The monitor is made of a measurement chamber surrounded by detectors and a shielded outer frame. The shielded chamber consists of a steel frame, which contains typically 1.5 inches (3.81 cm) of lead and 0.5 inches (1.27 cm) of steel shielding. Inside the shielding are plastic scintillator panels, which serve as gross gamma ray detectors. The detector panels, with embedded photomultipliers, completely surround the internal measurement chamber on all 6 sides. Care has been takenmore » to distribute the plastic scintillator detectors in order to optimise both the efficiency for gamma ray detection and at the same time achieve a volumetric sensitivity, which is as uniform as possible. A common high voltage power supply provides the bias voltage for each of the six photomultipliers. The voltage signals arising from the detectors and photomultipliers are amplified by six sensitive amplifiers. Each amplifier incorporates a single channel analyser with both upper and lower thresholds and the digitised counts from each detector are recorded on six scalars. Operation of the device is by means of a microprocessor from which the scalars are controlled. An internal load cell linked to the microprocessor determines the weight of the waste object, and this information is used to calculate the specific activity of the waste. The monitor makes background measurements when the shielded door is closed and a sample, usually a bag of low-density waste, is not present in the measurement chamber. Measurements of the minimum detectable activity (MDA) of an earlier large volume prototype instrument are reported as part of the development of the Waste Segregation and Clearance Monitor (WSCM) described in the paper. For the optimised WSCM a detection efficiency of greater than 32% was measured using a small Cs

  15. Approach to the vadose zone monitoring in hazardous and solid waste disposal facilities

    NASA Astrophysics Data System (ADS)

    Twardowska, Irena

    2004-03-01

    In the solid waste (SW)disposal sites, in particular at the unlined facilities, at the remediated or newly-constructed units equipped with novel protective/reactive permeable barriers or at lined facilities with leachate collection systems that are prone to failure, the vadose zone monitoring should comprise besides the natural soil layer beneath the landfill, also the anthropogenic vadose zone, i.e. the waste layer and pore solutions in the landfill. The vadose zone screening along the vertical profile of SW facilities with use of direct invasive soil-core and soil-pore liquid techniques shows vertical downward redistribution of inorganic (macroconstituents and heavy metals) and organic (PAHs) contaminant loads in water infiltrating through the waste layer. These loads can make ground water down-gradient of the dump unfit for any use. To avoid damage of protective/reactive permeable barriers and liners, an installation of stationary monitoring systems along the waste layer profile during the construction of a landfill, which are amenable to generate accurate data and information in a near-real time should be considered including:(i) permanent samplers of pore solution, with a periodic pump-induced transport of collected solution to the surface, preferably with instant field measurements;(ii)chemical sensors with continuous registration of critical parameters. These techniques would definitely provide an early alert in case when the chemical composition of pore solution percolating downward the waste profile shows unfavorable transformations, which indicate an excessive contaminant load approaching ground water. The problems concerning invasive and stationary monitoring of the vadose zone in SW disposal facilities will be discussed at the background of results of monitoring data and properties of permeable protective/reactive barriers considered for use.

  16. Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes.

    PubMed

    Klionsky, Daniel J; Abeliovich, Hagai; Agostinis, Patrizia; Agrawal, Devendra K; Aliev, Gjumrakch; Askew, David S; Baba, Misuzu; Baehrecke, Eric H; Bahr, Ben A; Ballabio, Andrea; Bamber, Bruce A; Bassham, Diane C; Bergamini, Ettore; Bi, Xiaoning; Biard-Piechaczyk, Martine; Blum, Janice S; Bredesen, Dale E; Brodsky, Jeffrey L; Brumell, John H; Brunk, Ulf T; Bursch, Wilfried; Camougrand, Nadine; Cebollero, Eduardo; Cecconi, Francesco; Chen, Yingyu; Chin, Lih-Shen; Choi, Augustine; Chu, Charleen T; Chung, Jongkyeong; Clarke, Peter G H; Clark, Robert S B; Clarke, Steven G; Clavé, Corinne; Cleveland, John L; Codogno, Patrice; Colombo, María I; Coto-Montes, Ana; Cregg, James M; Cuervo, Ana Maria; Debnath, Jayanta; Demarchi, Francesca; Dennis, Patrick B; Dennis, Phillip A; Deretic, Vojo; Devenish, Rodney J; Di Sano, Federica; Dice, J Fred; Difiglia, Marian; Dinesh-Kumar, Savithramma; Distelhorst, Clark W; Djavaheri-Mergny, Mojgan; Dorsey, Frank C; Dröge, Wulf; Dron, Michel; Dunn, William A; Duszenko, Michael; Eissa, N Tony; Elazar, Zvulun; Esclatine, Audrey; Eskelinen, Eeva-Liisa; Fésüs, László; Finley, Kim D; Fuentes, José M; Fueyo, Juan; Fujisaki, Kozo; Galliot, Brigitte; Gao, Fen-Biao; Gewirtz, David A; Gibson, Spencer B; Gohla, Antje; Goldberg, Alfred L; Gonzalez, Ramon; González-Estévez, Cristina; Gorski, Sharon; Gottlieb, Roberta A; Häussinger, Dieter; He, You-Wen; Heidenreich, Kim; Hill, Joseph A; Høyer-Hansen, Maria; Hu, Xun; Huang, Wei-Pang; Iwasaki, Akiko; Jäättelä, Marja; Jackson, William T; Jiang, Xuejun; Jin, Shengkan; Johansen, Terje; Jung, Jae U; Kadowaki, Motoni; Kang, Chanhee; Kelekar, Ameeta; Kessel, David H; Kiel, Jan A K W; Kim, Hong Pyo; Kimchi, Adi; Kinsella, Timothy J; Kiselyov, Kirill; Kitamoto, Katsuhiko; Knecht, Erwin; Komatsu, Masaaki; Kominami, Eiki; Kondo, Seiji; Kovács, Attila L; Kroemer, Guido; Kuan, Chia-Yi; Kumar, Rakesh; Kundu, Mondira; Landry, Jacques; Laporte, Marianne; Le, Weidong; Lei, Huan-Yao; Lenardo, Michael J; Levine, Beth; Lieberman, Andrew; Lim, Kah-Leong; Lin, Fu-Cheng; Liou, Willisa; Liu, Leroy F; Lopez-Berestein, Gabriel; López-Otín, Carlos; Lu, Bo; Macleod, Kay F; Malorni, Walter; Martinet, Wim; Matsuoka, Ken; Mautner, Josef; Meijer, Alfred J; Meléndez, Alicia; Michels, Paul; Miotto, Giovanni; Mistiaen, Wilhelm P; Mizushima, Noboru; Mograbi, Baharia; Monastyrska, Iryna; Moore, Michael N; Moreira, Paula I; Moriyasu, Yuji; Motyl, Tomasz; Münz, Christian; Murphy, Leon O; Naqvi, Naweed I; Neufeld, Thomas P; Nishino, Ichizo; Nixon, Ralph A; Noda, Takeshi; Nürnberg, Bernd; Ogawa, Michinaga; Oleinick, Nancy L; Olsen, Laura J; Ozpolat, Bulent; Paglin, Shoshana; Palmer, Glen E; Papassideri, Issidora; Parkes, Miles; Perlmutter, David H; Perry, George; Piacentini, Mauro; Pinkas-Kramarski, Ronit; Prescott, Mark; Proikas-Cezanne, Tassula; Raben, Nina; Rami, Abdelhaq; Reggiori, Fulvio; Rohrer, Bärbel; Rubinsztein, David C; Ryan, Kevin M; Sadoshima, Junichi; Sakagami, Hiroshi; Sakai, Yasuyoshi; Sandri, Marco; Sasakawa, Chihiro; Sass, Miklós; Schneider, Claudio; Seglen, Per O; Seleverstov, Oleksandr; Settleman, Jeffrey; Shacka, John J; Shapiro, Irving M; Sibirny, Andrei; Silva-Zacarin, Elaine C M; Simon, Hans-Uwe; Simone, Cristiano; Simonsen, Anne; Smith, Mark A; Spanel-Borowski, Katharina; Srinivas, Vickram; Steeves, Meredith; Stenmark, Harald; Stromhaug, Per E; Subauste, Carlos S; Sugimoto, Seiichiro; Sulzer, David; Suzuki, Toshihiko; Swanson, Michele S; Tabas, Ira; Takeshita, Fumihiko; Talbot, Nicholas J; Tallóczy, Zsolt; Tanaka, Keiji; Tanaka, Kozo; Tanida, Isei; Taylor, Graham S; Taylor, J Paul; Terman, Alexei; Tettamanti, Gianluca; Thompson, Craig B; Thumm, Michael; Tolkovsky, Aviva M; Tooze, Sharon A; Truant, Ray; Tumanovska, Lesya V; Uchiyama, Yasuo; Ueno, Takashi; Uzcátegui, Néstor L; van der Klei, Ida; Vaquero, Eva C; Vellai, Tibor; Vogel, Michael W; Wang, Hong-Gang; Webster, Paul; Wiley, John W; Xi, Zhijun; Xiao, Gutian; Yahalom, Joachim; Yang, Jin-Ming; Yap, George; Yin, Xiao-Ming; Yoshimori, Tamotsu; Yu, Li; Yue, Zhenyu; Yuzaki, Michisuke; Zabirnyk, Olga; Zheng, Xiaoxiang; Zhu, Xiongwei; Deter, Russell L

    2008-02-01

    Research in autophagy continues to accelerate,(1) and as a result many new scientists are entering the field. Accordingly, it is important to establish a standard set of criteria for monitoring macroautophagy in different organisms. Recent reviews have described the range of assays that have been used for this purpose.(2,3) There are many useful and convenient methods that can be used to monitor macroautophagy in yeast, but relatively few in other model systems, and there is much confusion regarding acceptable methods to measure macroautophagy in higher eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers of autophagosomes versus those that measure flux through the autophagy pathway; thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from fully functional autophagy that includes delivery to, and degradation within, lysosomes (in most higher eukaryotes) or the vacuole (in plants and fungi). Here, we present a set of guidelines for the selection and interpretation of the methods that can be used by investigators who are attempting to examine macroautophagy and related processes, as well as by reviewers who need to provide realistic and reasonable critiques of papers that investigate these processes. This set of guidelines is not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to verify an autophagic response.

  17. Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes

    PubMed Central

    Klionsky, Daniel J.; Abeliovich, Hagai; Agostinis, Patrizia; Agrawal, Devendra K.; Aliev, Gjumrakch; Askew, David S.; Baba, Misuzu; Baehrecke, Eric H.; Bahr, Ben A.; Ballabio, Andrea; Bamber, Bruce A.; Bassham, Diane C.; Bergamini, Ettore; Bi, Xiaoning; Biard-Piechaczyk, Martine; Blum, Janice S.; Bredesen, Dale E.; Brodsky, Jeffrey L.; Brumell, John H.; Brunk, Ulf T.; Bursch, Wilfried; Camougrand, Nadine; Cebollero, Eduardo; Cecconi, Francesco; Chen, Yingyu; Chin, Lih-Shen; Choi, Augustine; Chu, Charleen T.; Chung, Jongkyeong; Clarke, Peter G.H.; Clark, Robert S.B.; Clarke, Steven G.; Clavé, Corinne; Cleveland, John L.; Codogno, Patrice; Colombo, María I.; Coto-Montes, Ana; Cregg, James M.; Cuervo, Ana Maria; Debnath, Jayanta; Demarchi, Francesca; Dennis, Patrick B.; Dennis, Phillip A.; Deretic, Vojo; Devenish, Rodney J.; Di Sano, Federica; Dice, J. Fred; DiFiglia, Marian; Dinesh-Kumar, Savithramma; Distelhorst, Clark W.; Djavaheri-Mergny, Mojgan; Dorsey, Frank C.; Dröge, Wulf; Dron, Michel; Dunn, William A.; Duszenko, Michael; Eissa, N. Tony; Elazar, Zvulun; Esclatine, Audrey; Eskelinen, Eeva-Liisa; Fésüs, László; Finley, Kim D.; Fuentes, José M.; Fueyo, Juan; Fujisaki, Kozo; Galliot, Brigitte; Gao, Fen-Biao; Gewirtz, David A.; Gibson, Spencer B.; Gohla, Antje; Goldberg, Alfred L.; Gonzalez, Ramon; González-Estévez, Cristina; Gorski, Sharon; Gottlieb, Roberta A.; Häussinger, Dieter; He, You-Wen; Heidenreich, Kim; Hill, Joseph A.; Høyer-Hansen, Maria; Hu, Xun; Huang, Wei-Pang; Iwasaki, Akiko; Jäättelä, Marja; Jackson, William T.; Jiang, Xuejun; Jin, Shengkan; Johansen, Terje; Jung, Jae U.; Kadowaki, Motoni; Kang, Chanhee; Kelekar, Ameeta; Kessel, David H.; Kiel, Jan A.K.W.; Kim, Hong Pyo; Kimchi, Adi; Kinsella, Timothy J.; Kiselyov, Kirill; Kitamoto, Katsuhiko; Knecht, Erwin; Komatsu, Masaaki; Kominami, Eiki; Kondo, Seiji; Kovács, Attila L.; Kroemer, Guido; Kuan, Chia-Yi; Kumar, Rakesh; Kundu, Mondira; Landry, Jacques; Laporte, Marianne; Le, Weidong; Lei, Huan-Yao; Lenardo, Michael J.; Levine, Beth; Lieberman, Andrew; Lim, Kah-Leong; Lin, Fu-Cheng; Liou, Willisa; Liu, Leroy F.; Lopez-Berestein, Gabriel; López-Otín, Carlos; Lu, Bo; Macleod, Kay F.; Malorni, Walter; Martinet, Wim; Matsuoka, Ken; Mautner, Josef; Meijer, Alfred J.; Meléndez, Alicia; Michels, Paul; Miotto, Giovanni; Mistiaen, Wilhelm P.; Mizushima, Noboru; Mograbi, Baharia; Monastyrska, Iryna; Moore, Michael N.; Moreira, Paula I.; Moriyasu, Yuji; Motyl, Tomasz; Münz, Christian; Murphy, Leon O.; Naqvi, Naweed I.; Neufeld, Thomas P.; Nishino, Ichizo; Nixon, Ralph A.; Noda, Takeshi; Nürnberg, Bernd; Ogawa, Michinaga; Oleinick, Nancy L.; Olsen, Laura J.; Ozpolat, Bulent; Paglin, Shoshana; Palmer, Glen E.; Papassideri, Issidora; Parkes, Miles; Perlmutter, David H.; Perry, George; Piacentini, Mauro; Pinkas-Kramarski, Ronit; Prescott, Mark; Proikas-Cezanne, Tassula; Raben, Nina; Rami, Abdelhaq; Reggiori, Fulvio; Rohrer, Bärbel; Rubinsztein, David C.; Ryan, Kevin M.; Sadoshima, Junichi; Sakagami, Hiroshi; Sakai, Yasuyoshi; Sandri, Marco; Sasakawa, Chihiro; Sass, Miklós; Schneider, Claudio; Seglen, Per O.; Seleverstov, Oleksandr; Settleman, Jeffrey; Shacka, John J.; Shapiro, Irving M.; Sibirny, Andrei; Silva-Zacarin, Elaine C.M.; Simon, Hans-Uwe; Simone, Cristiano; Simonsen, Anne; Smith, Mark A.; Spanel-Borowski, Katharina; Srinivas, Vickram; Steeves, Meredith; Stenmark, Harald; Stromhaug, Per E.; Subauste, Carlos S.; Sugimoto, Seiichiro; Sulzer, David; Suzuki, Toshihiko; Swanson, Michele S.; Tabas, Ira; Takeshita, Fumihiko; Talbot, Nicholas J.; Tallóczy, Zsolt; Tanaka, Keiji; Tanaka, Kozo; Tanida, Isei; Taylor, Graham S.; Taylor, J. Paul; Terman, Alexei; Tettamanti, Gianluca; Thompson, Craig B.; Thumm, Michael; Tolkovsky, Aviva M.; Tooze, Sharon A.; Truant, Ray; Tumanovska, Lesya V.; Uchiyama, Yasuo; Ueno, Takashi; Uzcátegui, Néstor L.; van der Klei, Ida; Vaquero, Eva C.; Vellai, Tibor; Vogel, Michael W.; Wang, Hong-Gang; Webster, Paul; Wiley, John W.; Xi, Zhijun; Xiao, Gutian; Yahalom, Joachim; Yang, Jin-Ming; Yap, George; Yin, Xiao-Ming; Yoshimori, Tamotsu; Yu, Li; Yue, Zhenyu; Yuzaki, Michisuke; Zabirnyk, Olga; Zheng, Xiaoxiang; Zhu, Xiongwei; Deter, Russell L.

    2009-01-01

    Research in autophagy continues to accelerate,1 and as a result many new scientists are entering the field. Accordingly, it is important to establish a standard set of criteria for monitoring macroautophagy in different organisms. Recent reviews have described the range of assays that have been used for this purpose.2,3 There are many useful and convenient methods that can be used to monitor macroautophagy in yeast, but relatively few in other model systems, and there is much confusion regarding acceptable methods to measure macroautophagy in higher eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers of autophagosomes versus those that measure flux through the autophagy pathway; thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from fully functional autophagy that includes delivery to, and degradation within, lysosomes (in most higher eukaryotes) or the vacuole (in plants and fungi). Here, we present a set of guidelines for the selection and interpretation of the methods that can be used by investigators who are attempting to examine macroautophagy and related processes, as well as by reviewers who need to provide realistic and reasonable critiques of papers that investigate these processes. This set of guidelines is not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to verify an autophagic response. PMID:18188003

  18. Automated locomotor activity monitoring as a quality control assay for mass-reared tephritid flies.

    PubMed

    Dominiak, Bernard C; Fanson, Benjamin G; Collins, Samuel R; Taylor, Phillip W

    2014-02-01

    The Sterile Insect Technique (SIT) requires vast numbers of consistently high quality insects to be produced over long periods. Quality control (QC) procedures are critical to effective SIT, both providing quality assurance and warning of operational deficiencies. We here present a potential new QC assay for mass rearing of Queensland fruit flies (Bactrocera tryoni Froggatt) for SIT; locomotor activity monitoring. We investigated whether automated locomotor activity monitors (LAMs) that simply detect how often a fly passes an infrared sensor in a glass tube might provide similar insights but with much greater economy. Activity levels were generally lower for females than for males, and declined over five days in the monitor for both sexes. Female activity levels were not affected by irradiation, but males irradiated at 60 or 70 Gy had reduced activity levels compared with unirradiated controls. We also found some evidence that mild heat shock of pupae results in adults with reduced activity. LAM offers a convenient, effective and economical assay to probe such changes. © 2013 Society of Chemical Industry.

  19. Monitoring of anticoagulant therapy in heart disease: considerations for the current assays.

    PubMed

    Boroumand, Mohammadali; Goodarzynejad, Hamidreza

    2010-01-01

    Clinicians should be aware of new developments to familiarize themselves with pharmacokinetic and pharmacodynamic characteristics of new anticoagulant agents to appropriately and safely use them. For the moment, cardiologists and other clinicians also require to master currently available drugs, realizing the mechanism of action, side effects, and laboratory monitoring to measure their anticoagulant effects. Warfarin and heparin have narrow therapeutic window with high inter- and intra-patient variability, thereby the use of either drug needs careful laboratory monitoring and dose adjustment to ensure proper antithrombotic protection while minimizing the bleeding risk. The prothrombin time (PT) and the activated partial thromboplastin time (aPTT) are laboratory tests commonly used to monitor warfarin and heparin, respectively. These two tests depend highly on the combination of reagent and instrument utilized. Results for a single specimen tested in different laboratories are variable; this is mostly attributable to the specific reagents and to a much lesser degree to the instrument used. The PT stands alone as the single coagulation test that has undergone the most extensive attempt at assay standardization. The international normalized ratio (INR) was introduced to "normalize" all PT reagents to a World Health Organization (WHO) reference thromboplastin preparation standard, such that a PT measured anywhere in the world would result in an INR value similar to that which would have been achieved had the WHO reference thromboplastin been utilized. However, INRs are reproducible between laboratories for only those patients who are stably anticoagulated with vitamin K antagonists (VKAs) (i.e., at least 6 weeks of VKA therapy), and are not reliable or reproducible between laboratories for patients for whom VKA therapy has recently been started or any other clinical conditions associated with a prolonged PT such as liver disease, disseminated intravascular coagulation

  20. Hydrologic monitoring of a waste-injection well near Milton, Florida, June 1975 - December 1976

    USGS Publications Warehouse

    Pascale, Charles A.; Martin, J.B.

    1977-01-01

    Hydraulic and chemical data were collected through a monitoring program conducted by the U.S. Geological Survey at an industrial liquid-waste injection site 6 mi southwest of Milton, Fla., in Santa Rosa County. The injection system is described. Data include injection rates, volumes, and pressures; water-level data at three monitor wells and a standby injection well, and field and laboratory analyses of water samples from four wells. Hydraulic and geochemical effects of the waste-injection system at the plant as of December 31, 1976, have been detected only in the injection zone, the lower limestone of the Floridan aquifer. Increased pressures are evident at the three wells used to monitor the injection zone. Geochemical changes have been noted only at the deep-test monitor well closest to the injection well. (Woodard-USGS)

  1. Assay optimisation and technology transfer for multi-site immuno-monitoring in vaccine trials

    PubMed Central

    Harris, Stephanie A.; Satti, Iman; Bryan, Donna; Walker, K. Barry; Dockrell, Hazel M.; McShane, Helen; Ho, Mei Mei

    2017-01-01

    Cellular immunological assays are important tools for the monitoring of responses to T-cell-inducing vaccine candidates. As these bioassays are often technically complex and require considerable experience, careful technology transfer between laboratories is critical if high quality, reproducible data that allows comparison between sites, is to be generated. The aim of this study, funded by the European Union Framework Program 7-funded TRANSVAC project, was to optimise Standard Operating Procedures and the technology transfer process to maximise the reproducibility of three bioassays for interferon-gamma responses: enzyme-linked immunosorbent assay (ELISA), ex-vivo enzyme-linked immunospot and intracellular cytokine staining. We found that the initial variability in results generated across three different laboratories reduced following a combination of Standard Operating Procedure harmonisation and the undertaking of side-by-side training sessions in which assay operators performed each assay in the presence of an assay ‘lead’ operator. Mean inter-site coefficients of variance reduced following this training session when compared with the pre-training values, most notably for the ELISA assay. There was a trend for increased inter-site variability at lower response magnitudes for the ELISA and intracellular cytokine staining assays. In conclusion, we recommend that on-site operator training is an essential component of the assay technology transfer process and combined with harmonised Standard Operating Procedures will improve the quality, reproducibility and comparability of data produced across different laboratories. These data may be helpful in ongoing discussions of the potential risk/benefit of centralised immunological assay strategies for large clinical trials versus decentralised units. PMID:29020010

  2. Monitoring of waste disposal in deep geological formations

    NASA Astrophysics Data System (ADS)

    German, V.; Mansurov, V.

    2003-04-01

    In the paper application of kinetic approach for description of rock failure process and waste disposal microseismic monitoring is advanced. On base of two-stage model of failure process the capability of rock fracture is proved. The requests to monitoring system such as real time mode of data registration and processing and its precision range are formulated. The method of failure nuclei delineation in a rock masses is presented. This method is implemented in a software program for strong seismic events forecasting. It is based on direct use of the fracture concentration criterion. The method is applied to the database of microseismic events of the North Ural Bauxite Mine. The results of this application, such as: efficiency, stability, possibility of forecasting rockburst are discussed.

  3. Enhancing anaerobic digestion of food waste through biochemical methane potential assays at different substrate: inoculum ratios.

    PubMed

    Hobbs, Shakira R; Landis, Amy E; Rittmann, Bruce E; Young, Michelle N; Parameswaran, Prathap

    2018-01-01

    Food waste has a high energy potential that can be converted into useful energy in the form of methane via anaerobic digestion. Biochemical Methane Potential assays (BMPs) were conducted to quantify the impacts on methane production of different ratios of food waste. Anaerobic digester sludge (ADS) was used as the inoculum, and BMPs were performed at food waste:inoculum ratios of 0.42, 1.42, and 3.0g chemical oxygen demand/g volatile solids (VS). The 1.42 ratio had the highest CH 4 -COD recovery: 90% of the initial total chemical oxygen demand (TCOD) was from food waste, followed by ratios 0.42 and 3.0 at 69% and 57%, respectively. Addition of food waste above 0.42 caused a lag time for CH 4 production that increased with higher ratios, which highlighted the negative impacts of overloading with food waste. The Gompertz equation was able to represent the results well, and it gave lag times of 0, 3.6 and 30days and maximum methane productions of 370, 910, and 1950mL for ratios 0.42, 1.42 and 3.0, respectively. While ratio 3.0 endured a long lag phase and low VSS destruction, ratio 1.42 achieved satisfactory results for all performance criteria. These results provide practical guidance on food-waste-to-inoculum ratios that can lead to optimizing methanogenic yield. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Comparison of Three Different Sensitive Assays for Hepatitis B Virus DNA in Monitoring of Responses to Antiviral Therapy

    PubMed Central

    Chan, Henry L. Y.; Leung, Nancy W. Y.; Lau, Tracy C. M.; Wong, May L.; Sung, Joseph J. Y.

    2000-01-01

    The aim of our study was to compare the performances of two new hepatitis B virus (HBV) DNA assays, a cross-linking assay (NAXCOR) and a hybrid-capture amplification assay (Digene), versus the widely used branched-DNA (bDNA) assay (Chiron) in the monitoring of HBV DNA levels during antiviral treatment. Serial serum samples from 12 chronically HBV infected patients undergoing a phase II trial of an antiviral drug, 2′,3′-dideoxy-5-fluoro-3′-thiacytidine (FTC), were studied. A total of 96 serum samples were tested for HBV DNA using the cross-linking, hybrid-capture amplification, and bDNA assays. In the comparison of the cross-linking and bDNA assays, concordant results were found in 77 (80.3%) samples, no significant difference was found between the median log10 HBV DNA levels (6.66 versus 7.17 meq/ml), and the results of the two assays were closely correlated (r = 0.95). In the comparison of the hybrid-capture amplification and bDNA assays, concordant results were found in 79 (82.3%) samples, no significant difference was found between the median log10 HBV DNA levels (6.98 versus 6.99 meq/ml), and the results of the two assays were closely correlated (r = 0.99). Six (6.3%) samples by the cross-linking assay and 10 (10.4%) samples by the bDNA assay required retesting because of unacceptably high within-run coefficients of variance. No sample required retesting in the hybrid-capture amplification assay according to the internal validation. In conclusion, the cross-linking and hybrid-capture amplification assays were as sensitive as the bDNA assay for HBV DNA detection and can be recommended for monitoring of HBV DNA levels during antiviral treatment. PMID:10970358

  5. Statistical characterization of multiple-reaction monitoring mass spectrometry (MRM-MS) assays for quantitative proteomics

    PubMed Central

    2012-01-01

    Multiple reaction monitoring mass spectrometry (MRM-MS) with stable isotope dilution (SID) is increasingly becoming a widely accepted assay for the quantification of proteins and peptides. These assays have shown great promise in relatively high throughput verification of candidate biomarkers. While the use of MRM-MS assays is well established in the small molecule realm, their introduction and use in proteomics is relatively recent. As such, statistical and computational methods for the analysis of MRM-MS data from proteins and peptides are still being developed. Based on our extensive experience with analyzing a wide range of SID-MRM-MS data, we set forth a methodology for analysis that encompasses significant aspects ranging from data quality assessment, assay characterization including calibration curves, limits of detection (LOD) and quantification (LOQ), and measurement of intra- and interlaboratory precision. We draw upon publicly available seminal datasets to illustrate our methods and algorithms. PMID:23176545

  6. Statistical characterization of multiple-reaction monitoring mass spectrometry (MRM-MS) assays for quantitative proteomics.

    PubMed

    Mani, D R; Abbatiello, Susan E; Carr, Steven A

    2012-01-01

    Multiple reaction monitoring mass spectrometry (MRM-MS) with stable isotope dilution (SID) is increasingly becoming a widely accepted assay for the quantification of proteins and peptides. These assays have shown great promise in relatively high throughput verification of candidate biomarkers. While the use of MRM-MS assays is well established in the small molecule realm, their introduction and use in proteomics is relatively recent. As such, statistical and computational methods for the analysis of MRM-MS data from proteins and peptides are still being developed. Based on our extensive experience with analyzing a wide range of SID-MRM-MS data, we set forth a methodology for analysis that encompasses significant aspects ranging from data quality assessment, assay characterization including calibration curves, limits of detection (LOD) and quantification (LOQ), and measurement of intra- and interlaboratory precision. We draw upon publicly available seminal datasets to illustrate our methods and algorithms.

  7. Optical characteristics of waste stabilization ponds: recommendations for monitoring.

    PubMed

    Davies-Colley, R J; Craggs, R J; Park, J; Nagels, J W

    2005-01-01

    The optical character of waste stabilization ponds (WSPs) is of concern for several reasons. Algal photosynthesis, which produces oxygen for waste oxidation in WSPs, is influenced by attenuation of sunlight in ponds. Disinfection in WSPs is influenced by optical characteristics because solar UV exposure usually dominates inactivation. The optical nature of WSPs effluent also affects assimilation by receiving waters. Despite the importance of light behaviour in WSPs, few studies have been made of their optical characteristics. We discuss simple optical measures suitable for routine monitoring of WSPs (including at sites remote from laboratories): optical density of filtrates - an index of dissolved coloured organic (humic) matter, visual clarity - to provide an estimate of the beam attenuation coefficient (a fundamental quantity needed for optical modelling) colour (hue) - as an indicator of general WSP 'condition' and irradiance attenuation quantifying depth of light penetration. The value of optical characterisation of WSPs is illustrated with reference to optical data for WSPs in NZ (including high-rate algal ponds) treating dairy cattle wastewater versus domestic sewage. We encourage increased research on optical characteristics of WSPs and the incorporation of optical measures in monitoring and modelling of WSP performance.

  8. Urban observatories opportunities for environmental monitoring: solid wastes.

    PubMed

    Rojas-Caldelas, R I; Corona Zambrano, E A

    2008-01-01

    Towns concentrate around 50% of world-wide population and the trend is oriented to underscore an urban profile of population. In addition, towns have become important for their economic contribution to the Gross Internal Product. The negative side of towns is the environmental and social impacts as a result of productive and domestic activities, besides the lack of available data. In order to overcome these shortcomings, the United Nations has established a project of urban monitoring throughout the Global Network of Urban Observatories; Mexico joined the project in 2005. The Local Urban Observatory of Mexicali has the task to produce information about cities that is useful to design public policies. Some of this information deals with a set of environmental indicators in the United Nations Habitat Agenda, which includes solid wastes. Therefore, this paper deals with two main topics; firstly, from the Habitat Agenda, a comparative urban analysis of waste production and coverage of domestic waste collection services; secondly, from the Local Agenda, the identification and ranking of environmental problems according to public perception coming from people involved in the municipal planning and decision making process. Results will be used to develop local indicators and public environmental policies.

  9. Project characteristics monitoring report: BWIP (Basalt Waste Isolation Program) repository project

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Friedli, E.A.; Herborn, D.I.; Taylor, C.D.

    1988-03-01

    This monitoring report has been prepared to show compliance with provisions of the Nuclear Waste Policy Act of 1982 (NWPA) and to provide local and state government agencies with information concerning the Basalt Waste Isolation Program (BWIP). This report contains data for the time period May 26, 1986 to February 1988. The data include employment figures, salaries, project purchases, taxes and fees paid, worker survey results, and project closedown personal interview summaries. This information has become particularly important since the decision in December 1987 to stop all BWIP activities except those for site reclamation. The Nuclear Waste Policy Amendments Actmore » of 1987 requires nonreclamation work at the Hanford Site to stop as of March 22, 1988. 7 refs., 6 figs., 28 tabs.« less

  10. Efficacy of T2 Magnetic Resonance Assay in Monitoring Candidemia after Initiation of Antifungal Therapy: the Serial Therapeutic and Antifungal Monitoring Protocol (STAMP) Trial

    PubMed Central

    2018-01-01

    ABSTRACT The performance of blood culture for monitoring candidemia clearance is hampered by its low sensitivity, especially during antifungal therapy. The T2 magnetic resonance (T2MR) assay combines magnetic resonance with nanotechnology to identify whole Candida species cells. A multicenter clinical trial studied the performance of T2MR in monitoring candidemia clearance compared to blood culture. Adults with a blood culture positive for yeast were enrolled and had blood cultures and T2MR testing performed on prespecified days. Thirty-one patients completed the trial. Thirteen of the 31 patients (41.9%) had at least one positive surveillance T2MR and/or blood culture result. All positive blood cultures (7/7 [100%]) had an accompanying positive T2MR result with concordance in the identified Candida sp., while only 7/23 (30.4%) T2MR results had an accompanying positive blood culture. There was one case of discordance in species identification between T2MR and the preenrollment blood culture with evidence to support deep-seated infection by the Candida spp. detected by the T2MR assay. Based on the log rank test, there was a statistically significant improvement in posttreatment surveillance using the T2MR assay compared to blood culture (P = 0.004). Limitations of the study include the small sample size and lack of outcome data. In conclusion, the T2MR assay significantly outperformed blood cultures for monitoring the clearance of candidemia in patients receiving antifungal therapy and may be useful in determining adequate source control, timing for deescalation, and optimal duration of treatment. However, further studies are needed to determine the viability of Candida species cells detected by the T2MR assay and correlate the results with patient outcomes. (This study is registered at ClinicalTrials.gov under registration number NCT02163889.) PMID:29367293

  11. Efficacy of T2 Magnetic Resonance Assay in Monitoring Candidemia after Initiation of Antifungal Therapy: the Serial Therapeutic and Antifungal Monitoring Protocol (STAMP) Trial.

    PubMed

    Mylonakis, Eleftherios; Zacharioudakis, Ioannis M; Clancy, Cornelius J; Nguyen, M Hong; Pappas, Peter G

    2018-04-01

    The performance of blood culture for monitoring candidemia clearance is hampered by its low sensitivity, especially during antifungal therapy. The T2 magnetic resonance (T2MR) assay combines magnetic resonance with nanotechnology to identify whole Candida species cells. A multicenter clinical trial studied the performance of T2MR in monitoring candidemia clearance compared to blood culture. Adults with a blood culture positive for yeast were enrolled and had blood cultures and T2MR testing performed on prespecified days. Thirty-one patients completed the trial. Thirteen of the 31 patients (41.9%) had at least one positive surveillance T2MR and/or blood culture result. All positive blood cultures (7/7 [100%]) had an accompanying positive T2MR result with concordance in the identified Candida sp., while only 7/23 (30.4%) T2MR results had an accompanying positive blood culture. There was one case of discordance in species identification between T2MR and the preenrollment blood culture with evidence to support deep-seated infection by the Candida spp. detected by the T2MR assay. Based on the log rank test, there was a statistically significant improvement in posttreatment surveillance using the T2MR assay compared to blood culture ( P = 0.004). Limitations of the study include the small sample size and lack of outcome data. In conclusion, the T2MR assay significantly outperformed blood cultures for monitoring the clearance of candidemia in patients receiving antifungal therapy and may be useful in determining adequate source control, timing for deescalation, and optimal duration of treatment. However, further studies are needed to determine the viability of Candida species cells detected by the T2MR assay and correlate the results with patient outcomes. (This study is registered at ClinicalTrials.gov under registration number NCT02163889.). Copyright © 2018 Mylonakis et al.

  12. Factor Activity Assays for Monitoring Extended Half-Life FVIII and Factor IX Replacement Therapies.

    PubMed

    Kitchen, Steve; Tiefenbacher, Stefan; Gosselin, Robert

    2017-04-01

    The advent of modified factor VIII (FVIII) and factor IX (FIX) molecules with extended half-lives (EHLs) compared with native FVIII and FIX represents a major advance in the field of hemophilia care, with the potential to reduce the frequency of prophylactic injections and/or to increase the trough level prior to subsequent injections. Monitoring treatment through laboratory assays will be an important part of ensuring patient safety, including any tailoring of prophylaxis. Several approaches have been used to extend half-lives, including PEGylation, and fusion to albumin or immunoglobulin. Some of these modifications affect factor assays as routinely performed in hemophilia centers; so, laboratories will need to use FVIII and FIX assays which have been shown to be suitable on a product-by-product basis. For some products, there are marked differences between results obtained using one-stage or chromogenic assays and results obtained using different reagents in the one-stage assay. The laboratory should use an assay in which the recovery of the product closely aligns with the assay used by the pharmaceutical company to assign potency to the product, so that the units reported by the laboratory agree with those used to demonstrate efficacy of the product during clinical trials. Reported assay differences in relation to several of the EHL FVIII and FIX molecules will be reviewed in this article. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

  13. Dealing with electronic waste: modeling the costs and environmental benefits of computer monitor disposal.

    PubMed

    Macauley, Molly; Palmer, Karen; Shih, Jhih-Shyang

    2003-05-01

    The importance of information technology to the world economy has brought about a surge in demand for electronic equipment. With rapid technological change, a growing fraction of the increasing stock of many types of electronics becomes obsolete each year. We model the costs and benefits of policies to manage 'e-waste' by focusing on a large component of the electronic waste stream-computer monitors-and the environmental concerns associated with disposal of the lead embodied in cathode ray tubes (CRTs) used in most monitors. We find that the benefits of avoiding health effects associated with CRT disposal appear far outweighed by the costs for a wide range of policies. For the stock of monitors disposed of in the United States in 1998, we find that policies restricting or banning some popular disposal options would increase disposal costs from about US dollar 1 per monitor to between US dollars 3 and US dollars 20 per monitor. Policies to promote a modest amount of recycling of monitor parts, including lead, can be less expensive. In all cases, however, the costs of the policies exceed the value of the avoided health effects of CRT disposal.

  14. FEMOS - Advanced Neutron Monitor System for Waste Management

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Simon, G.G.; Sokcic-Kostic, M.

    2006-07-01

    FEMOS is a specially developed monitor to detect fissile materials for waste characterisation and it is also suitable to identify the main neutron emitters. The latest measuring prototype is in routine operation at FZK-HDB (Germany) for determining Plutonium content and alpha activity in 2001 and 4001 drums from the reprocessing plant Karlsruhe (WAK). Authorities license the measurement procedure. Routine measurements conducted over a period of about 6 years and the results show that all specified prerequisites are met with the required reliability. (authors)

  15. Adaptation and evaluation of the bottle assay for monitoring insecticide resistance in disease vector mosquitoes in the Peruvian Amazon

    PubMed Central

    Zamora Perea, Elvira; Balta León, Rosario; Palomino Salcedo, Miriam; Brogdon, William G; Devine, Gregor J

    2009-01-01

    Background The purpose of this study was to establish whether the "bottle assay", a tool for monitoring insecticide resistance in mosquitoes, can complement and augment the capabilities of the established WHO assay, particularly in resource-poor, logistically challenging environments. Methods Laboratory reared Aedes aegypti and field collected Anopheles darlingi and Anopheles albimanus were used to assess the suitability of locally sourced solvents and formulated insecticides for use with the bottle assay. Using these adapted protocols, the ability of the bottle assay and the WHO assay to discriminate between deltamethrin-resistant Anopheles albimanus populations was compared. The diagnostic dose of deltamethrin that would identify resistance in currently susceptible populations of An. darlingi and Ae. aegypti was defined. The robustness of the bottle assay during a surveillance exercise in the Amazon was assessed. Results The bottle assay (using technical or formulated material) and the WHO assay were equally able to differentiate deltamethrin-resistant and susceptible An. albimanus populations. A diagnostic dose of 10 μg a.i./bottle was identified as the most sensitive discriminating dose for characterizing resistance in An. darlingi and Ae. aegypti. Treated bottles, prepared using locally sourced solvents and insecticide formulations, can be stored for > 14 days and used three times. Bottles can be stored and transported under local conditions and field-assays can be completed in a single evening. Conclusion The flexible and portable nature of the bottle assay and the ready availability of its components make it a potentially robust and useful tool for monitoring insecticide resistance and efficacy in remote areas that require minimal cost tools. PMID:19728871

  16. On-line remote monitoring of radioactive waste repositories

    NASA Astrophysics Data System (ADS)

    Calì, Claudio; Cosentino, Luigi; Litrico, Pietro; Pappalardo, Alfio; Scirè, Carlotta; Scirè, Sergio; Vecchio, Gianfranco; Finocchiaro, Paolo; Alfieri, Severino; Mariani, Annamaria

    2014-12-01

    A low-cost array of modular sensors for online monitoring of radioactive waste was developed at INFN-LNS. We implemented a new kind of gamma counter, based on Silicon PhotoMultipliers and scintillating fibers, that behaves like a cheap scintillating Geiger-Muller counter. It can be placed in shape of a fine grid around each single waste drum in a repository. Front-end electronics and an FPGA-based counting system were developed to handle the field data, also implementing data transmission, a graphical user interface and a data storage system. A test of four sensors in a real radwaste storage site was performed with promising results. Following the tests an agreement was signed between INFN and Sogin for the joint development and installation of a prototype DMNR (Detector Mesh for Nuclear Repository) system inside the Garigliano radwaste repository in Sessa Aurunca (CE, Italy). Such a development is currently under way, with the installation foreseen within 2014.

  17. (1→3)-β-D-Glucan Assay in Monitoring Response to Anti-Fungal Therapy in Fungal Endocarditis.

    PubMed

    Slim, Jihad; Saling, Christopher; Szabela, Maria; Brown, Melinda; Johnson, Tamara; Goldfarb, Irvin

    2017-03-01

    A case is reported of Candida glabrata infective endocarditis (IE) treated without surgical intervention. The study aim was to: (i) briefly discuss the outcomes of other documented cases of fungal IE managed medically with fluconazole; (ii) discuss the (1→3)-β-D-glucan assay and its previously studied role in the diagnosis of invasive fungal infections; and (iii) examine a possible application of the (1→3)-β-D-glucan assay to monitor response to antifungal treatment in patients with Candida endocarditis. The serum Fungitell assay was used to trend (1→3)-β-D-glucan in a patient with Candida endocarditis to determine treatment effectiveness with fluconazole, to provide an appropriate end date for antifungal therapy, and to survey infection suppression while off treatment. The (1→03)-β-D-glucan assay began trending downwards at 197 days into treatment with oral fluconazole. After 16 months of therapy, fluconazole was stopped due to transaminitis. (1→3)-β-Dglucan levels were checked six weeks after the discontinuation of treatment and were negative. The patient has now been off therapy for 21 weeks with no signs of clinical disease, and values remain negative. The present case indicates that a trending (1→3)-β-D-glucan assay may have valuable application in monitoring treatment response and infection suppression for Candida endocarditis.

  18. 40 CFR Table 11 to Subpart G of... - Wastewater-Inspection and Monitoring Requirements for Waste Management Units

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 9 2011-07-01 2011-07-01 false Wastewater-Inspection and Monitoring... and Monitoring Requirements for Waste Management Units To comply with Inspection or monitoring requirement Frequency of inspection or monitoring Method Tanks: 63.133(b)(1) Inspect fixed roof and all...

  19. 40 CFR Table 11 to Subpart G of... - Wastewater-Inspection and Monitoring Requirements for Waste Management Units

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 10 2012-07-01 2012-07-01 false Wastewater-Inspection and Monitoring... and Monitoring Requirements for Waste Management Units To comply with Inspection or monitoring requirement Frequency of inspection or monitoring Method Tanks: 63.133(b)(1) Inspect fixed roof and all...

  20. 40 CFR Table 11 to Subpart G of... - Wastewater-Inspection and Monitoring Requirements for Waste Management Units

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 10 2014-07-01 2014-07-01 false Wastewater-Inspection and Monitoring... and Monitoring Requirements for Waste Management Units To comply with Inspection or monitoring requirement Frequency of inspection or monitoring Method Tanks: 63.133(b)(1) Inspect fixed roof and all...

  1. 40 CFR Table 11 to Subpart G of... - Wastewater-Inspection and Monitoring Requirements for Waste Management Units

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 10 2013-07-01 2013-07-01 false Wastewater-Inspection and Monitoring... and Monitoring Requirements for Waste Management Units To comply with Inspection or monitoring requirement Frequency of inspection or monitoring Method Tanks: 63.133(b)(1) Inspect fixed roof and all...

  2. Lectin binding assays for in-process monitoring of sialylation in protein production.

    PubMed

    Xu, Weiduan; Chen, Jianmin; Yamasaki, Glenn; Murphy, John E; Mei, Baisong

    2010-07-01

    Many therapeutic proteins require appropriate glycosylation for their biological activities and plasma half life. Coagulation factor VIII (FVIII) is a glycoprotein which has extensive post-translational modification by N-linked glycosylation. The terminal sialic acid in the N-linked glycans of FVIII is required for maximal circulatory half life. The extent of FVIII sialylation can be determined by high pH anion-exchange chromatography coupled with a pulse electrochemical detector (HPAEC-PED), but this requires a large amount of purified protein. Using FVIII as a model, the objective of the present study was to develop assays that enable detection and prediction of sialylation deficiency at an early stage in the process and thus prevent downstream product quality excursions. Lectin ECA (Erythrina Cristagalli) binds to unsialylated Galbeta1-4 GlcNAc and the ECA-binding level (i.e., terminal Gal(beta1-4) exposure) is inversely proportional to the level of sialylation. By using ECA, a cell-based assay was developed to measure the global sialylation profile in FVIII producing cells. To examine the Galbeta1-4 exposure on the FVIII molecule in bioreactor tissue culture fluid (TCF), an ELISA-based ECA-FVIII binding assay was developed. The ECA-binding specificity in both assays was assessed by ECA-specific sugar inhibitors and neuraminidase digestion. The ECA-binding specificity was also independently confirmed by a ST3GAL4 siRNA knockdown experiment. To establish the correlation between Galbeta1-4 exposure and the HPAEC-PED determined FVIII sialylation value, the FVIII containing bioreactor TCF and the purified FVIII samples were tested with ECA ELISA binding assay. The results indicated an inverse correlation between ECA binding and the corresponding HPAEC-PED sialylation value. The ECA-binding assays are cost effective and can be rapidly performed, thereby making them effective for in-process monitoring of protein sialylation.

  3. Rapid anti-transglutaminase assay and patient interview for monitoring dietary compliance in celiac disease.

    PubMed

    Zanchi, Chiara; Ventura, Alessandro; Martelossi, Stefano; Di Leo, Grazia; Di Toro, Nicola; Not, Tarcisio

    2013-06-01

    The anti-transglutaminase antibodies (anti-tTG) play an important role in monitoring the celiacs' gluten-free diet (GFD). The authors propose to use the rapid IgA anti-tTG assay based on a whole blood drop to evaluate the compliance to GFD at the clinical ambulatory setting. The rapid test results were compared with those of the conventional ELISA assay and with dietary compliance reported by patients' interview. The authors showed that anti-tTG rapid test is reliable and easy to perform in the ambulatory setting to evaluate dietary compliance. Moreover, they proved that celiacs' interview is more sensitive than serology in identifying patients who transgress.

  4. A novel dual luciferase assay for the simultaneous monitoring of HIV infection and cell viability.

    PubMed

    Mitsuki, Yu-Ya; Yamamoto, Takuya; Mizukoshi, Fuminori; Momota, Masatoshi; Terahara, Kazutaka; Yoshimura, Kazuhisa; Harada, Shigeyoshi; Tsunetsugu-Yokota, Yasuko

    2016-05-01

    Human immunodeficiency virus type 1 (HIV-1) reporter cell lines are critical tools for drug development. However, one disadvantage of HIV-1 reporter cell lines is that reductions in reporter gene activity need to be normalized to cytotoxicity, i.e., live cell numbers. Here, we developed a dual luciferase assay based on a R. reniformis luciferase (hRLuc)-expressing R5-type HIV-1 (NLAD8-hRLuc) and a CEM cell line expressing CCR5 and firefly luciferase (R5CEM-FiLuc). The NLAD8-hRLuc reporter virus was replication competent in peripheral blood mononuclear cells. The level of hRLuc was correlated with p24 antigen levels (p<0.001, R=0.862). The target cell line, R5CEM-FiLuc, stably expressed the firefly luciferase (FiLuc) reporter gene and allowed the simultaneous monitoring of compound cytotoxicity. The dual reporter assay combining a NLAD8-hRLuc virus with R5CEM-FiLuc cells permitted the accurate determination of drug susceptibility for entry, reverse transcriptase, integrase, and protease inhibitors at different multiplicities of infection. This dual reporter assay provides a rapid and direct method for the simultaneous monitoring of HIV infection and cell viability. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Arsenic and lead contamination in urban soils of Villa de la Paz (Mexico) affected by historical mine wastes and its effect on children's health studied by micronucleated exfoliated cells assay.

    PubMed

    Gamiño-Gutiérrez, Sandra P; González-Pérez, C Ivonne; Gonsebatt, María E; Monroy-Fernández, Marcos G

    2013-02-01

    Environmental geochemical and health studies were carried out in urban areas of Villa de la Paz, S.L.P. (Mexico), where mining activities have been developed for more of 200 years, leading to the pollution of surface soil by arsenic and heavy metals (Pb, Cd, Cu, Zn). The analysis of urban soils to determine total and bioaccessibility concentrations of As and Pb, demonstrated a combined contribution of the natural and anthropogenic concentrations in the site, at levels higher than the environmental guideline values that provoke a human health risk. Contour soil mapping confirmed that historical mine waste deposits without environmental control measures, are the main source of pollution soil by As and Pb in the site. Exposure (Pb in blood and As in urine) and effect (micronucleated exfoliated cells assay) biological monitoring were then carried out in the childhood population of the site and in a control site. The exposure biological monitoring demonstrated that at least 20-30 % of children presented Pb and As exposure values higher than the national and international maximum intervention values. The effect biomonitoring by MEC assay confirmed that there is a genotoxic damage in local childhood population that could be associated with the arsenic exposure in the site.

  6. NRC Monitoring of Salt Waste Disposal at the Savannah River Site - 13147

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pinkston, Karen E.; Ridge, A. Christianne; Alexander, George W.

    2013-07-01

    As part of monitoring required under Section 3116 of the Ronald W. Reagan National Defense Authorization Act for Fiscal Year 2005 (NDAA), the NRC staff reviewed an updated DOE performance assessment (PA) for salt waste disposal at the Saltstone Disposal Facility (SDF). The NRC staff concluded that it has reasonable assurance that waste disposal at the SDF meets the 10 CFR 61 performance objectives for protection of individuals against intrusion (chap.61.42), protection of individuals during operations (chap.61.43), and site stability (chap.61.44). However, based on its evaluation of DOE's results and independent sensitivity analyses conducted with DOE's models, the NRC staffmore » concluded that it did not have reasonable assurance that DOE's disposal activities at the SDF meet the performance objective for protection of the general population from releases of radioactivity (chap.61.41) evaluated at a dose limit of 0.25 mSv/yr (25 mrem/yr) total effective dose equivalent (TEDE). NRC staff also concluded that the potential dose to a member of the public is expected to be limited (i.e., is expected to be similar to or less than the public dose limit in chap.20.1301 of 1 mSv/yr [100 mrem/yr] TEDE) and is expected to occur many years after site closure. The NRC staff used risk insights gained from review of the SDF PA, its experience monitoring DOE disposal actions at the SDF over the last 5 years, as well as independent analysis and modeling to identify factors that are important to assessing whether DOE's disposal actions meet the performance objectives. Many of these factors are similar to factors identified in the NRC staff's 2005 review of salt waste disposal at the SDF. Key areas of interest continue to be waste form and disposal unit degradation, the effectiveness of infiltration and erosion controls, and estimation of the radiological inventory. Based on these factors, NRC is revising its plan for monitoring salt waste disposal at the SDF in coordination with

  7. 2015 Groundwater Radiological Monitoring Results Associated with the Advanced Test Reactor Complex Cold Waste Ponds

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lewis, Michael George

    This report summarizes radiological monitoring results from groundwater wells associated with the Idaho National Laboratory Site’s Advanced Test Reactor Complex Cold Waste Ponds Reuse Permit (I-161-02). All radiological monitoring is performed to fulfill Department of Energy requirements under the Atomic Energy Act.

  8. An Electrochemical Impedance Spectroscopy System for Monitoring Pineapple Waste Saccharification.

    PubMed

    Conesa, Claudia; Ibáñez Civera, Javier; Seguí, Lucía; Fito, Pedro; Laguarda-Miró, Nicolás

    2016-02-04

    Electrochemical impedance spectroscopy (EIS) has been used for monitoring the enzymatic pineapple waste hydrolysis process. The system employed consists of a device called Advanced Voltammetry, Impedance Spectroscopy & Potentiometry Analyzer (AVISPA) equipped with a specific software application and a stainless steel double needle electrode. EIS measurements were conducted at different saccharification time intervals: 0, 0.75, 1.5, 6, 12 and 24 h. Partial least squares (PLS) were used to model the relationship between the EIS measurements and the sugar determination by HPAEC-PAD. On the other hand, artificial neural networks: (multilayer feed forward architecture with quick propagation training algorithm and logistic-type transfer functions) gave the best results as predictive models for glucose, fructose, sucrose and total sugars. Coefficients of determination (R²) and root mean square errors of prediction (RMSEP) were determined as R² > 0.944 and RMSEP < 1.782 for PLS and R² > 0.973 and RMSEP < 0.486 for artificial neural networks (ANNs), respectively. Therefore, a combination of both an EIS-based technique and ANN models is suggested as a promising alternative to the traditional laboratory techniques for monitoring the pineapple waste saccharification step.

  9. An Electrochemical Impedance Spectroscopy System for Monitoring Pineapple Waste Saccharification

    PubMed Central

    Conesa, Claudia; Ibáñez Civera, Javier; Seguí, Lucía; Fito, Pedro; Laguarda-Miró, Nicolás

    2016-01-01

    Electrochemical impedance spectroscopy (EIS) has been used for monitoring the enzymatic pineapple waste hydrolysis process. The system employed consists of a device called Advanced Voltammetry, Impedance Spectroscopy & Potentiometry Analyzer (AVISPA) equipped with a specific software application and a stainless steel double needle electrode. EIS measurements were conducted at different saccharification time intervals: 0, 0.75, 1.5, 6, 12 and 24 h. Partial least squares (PLS) were used to model the relationship between the EIS measurements and the sugar determination by HPAEC-PAD. On the other hand, artificial neural networks: (multilayer feed forward architecture with quick propagation training algorithm and logistic-type transfer functions) gave the best results as predictive models for glucose, fructose, sucrose and total sugars. Coefficients of determination (R2) and root mean square errors of prediction (RMSEP) were determined as R2 > 0.944 and RMSEP < 1.782 for PLS and R2 > 0.973 and RMSEP < 0.486 for artificial neural networks (ANNs), respectively. Therefore, a combination of both an EIS-based technique and ANN models is suggested as a promising alternative to the traditional laboratory techniques for monitoring the pineapple waste saccharification step. PMID:26861317

  10. 40 CFR 60.3067 - How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... curtain incinerators that burn only wood waste, clean lumber, and yard waste? 60.3067 Section 60.3067... Incinerators That Burn Only Wood Waste, Clean Lumber, and Yard Waste § 60.3067 How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber, and yard waste? (a) Use Method 9 of...

  11. 40 CFR 60.3067 - How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... curtain incinerators that burn only wood waste, clean lumber, and yard waste? 60.3067 Section 60.3067... Incinerators That Burn Only Wood Waste, Clean Lumber, and Yard Waste § 60.3067 How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber, and yard waste? (a) Use Method 9 of...

  12. 40 CFR 60.3067 - How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... curtain incinerators that burn only wood waste, clean lumber, and yard waste? 60.3067 Section 60.3067... Incinerators That Burn Only Wood Waste, Clean Lumber, and Yard Waste § 60.3067 How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber, and yard waste? (a) Use Method 9 of...

  13. 40 CFR 60.3067 - How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... curtain incinerators that burn only wood waste, clean lumber, and yard waste? 60.3067 Section 60.3067... Incinerators That Burn Only Wood Waste, Clean Lumber, and Yard Waste § 60.3067 How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber, and yard waste? (a) Use Method 9 of...

  14. 40 CFR 60.3067 - How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... curtain incinerators that burn only wood waste, clean lumber, and yard waste? 60.3067 Section 60.3067... Incinerators That Burn Only Wood Waste, Clean Lumber, and Yard Waste § 60.3067 How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber, and yard waste? (a) Use Method 9 of...

  15. Gas chromatography-mass spectrometry assay method for the therapeutic drug monitoring of the antiepileptic drug tiagabine.

    PubMed

    Chollet, D F; Castella, E; Goumaz, L; Anderegg, G

    1999-11-01

    A gas chromatography-mass spectrometry assay method suitable for the therapeutic drug monitoring of the antiepileptic drug tiagabine is described. Tiagabine and its desmethylated analogue used as internal standard were first extracted from serum by liquid-liquid extraction using an ethyl ether-isobutanol 98:2 mixture. Tiagabine and the internal standard were then methylated in the organic phase in presence of methanol by means of a safe and stable diazomethane derivative. After evaporation, the reconstituted extracts were chromatographed on a crosslinked phenyl methyl siloxane capillary column and detected by mass fragmentometry at m/z = 156. No other antiepileptic drug possibly administrated in polytherapy and no metabolite were found to interfere in the assay. The limit of quantification was 5 ng/ml. The precision and the accuracy were found to be suitable for the therapeutic drug monitoring of tiagabine.

  16. [Conservative calibration of a clearance monitor system for waste material from nuclear medicine].

    PubMed

    Wanke, Carsten; Geworski, Lilli

    2014-09-01

    Clearance monitor systems are used for gross gamma measurements of waste potentially contaminated with radioactivity. These measurements are to make sure that legal requirements, e.g. clearance criteria according to the german radiation protection ordinance, are met. This means that measurement results may overestimate, but must not underestimate the true values. This paper describes a pragmatic way using a calibrated Cs-137 point source to generate a conservative calibration for the clearance monitor system used in the Medizinische Hochschule Hannover (MHH). The most important nuclides used in nuclear medicine are considered. The measurement result reliably overestimates the true value of the activity present in the waste. The calibration is compliant with the demands for conservativity and traceability to national standards. Copyright © 2014. Published by Elsevier GmbH.

  17. EVALUATION OF AN ENZYME-LINKED IMMUNOSORBENT ASSAY FOR BIOLOGICAL MONITORING OF 3-PHENOXYBENZOIC ACID IN URINE

    EPA Science Inventory

    Abstract describes the development of an enzyme-linked immunosorbent assay (ELISA) method for monitoring 2,4-dichlorophenoxyacetic acid (2,4-D exposures). The ELISA is compared with a gas chromatograhy/mass spectrometry procedure. ELISA method development steps and comparative ...

  18. 40 CFR 60.2972 - How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... curtain incinerators that burn only wood waste, clean lumber, and yard waste? 60.2972 Section 60.2972... Only Wood Waste, Clean Lumber, and Yard Waste § 60.2972 How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber, and yard waste? (a) Use Method 9 of appendix A of this...

  19. 40 CFR 60.2972 - How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... curtain incinerators that burn only wood waste, clean lumber, and yard waste? 60.2972 Section 60.2972... Only Wood Waste, Clean Lumber, and Yard Waste § 60.2972 How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber, and yard waste? (a) Use Method 9 of appendix A of this...

  20. Monitoring genetic damage to ecosystems from hazardous waste

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Anderson, S.L.

    1992-03-01

    Applications of ecological toxicity testing to hazardous waste management have increased dramatically over the last few years, resulting in a greater awareness of the need for improved biomonitoring techniques. Our laboratory is developing advanced techniques to assess the genotoxic effects of environmental contamination on ecosystems. We have developed a novel mutagenesis assay using the nematode Caenorhabditis elegans, which is potentially applicable for multimedia studies in soil, sediment, and water. In addition, we are conducting validation studies of a previously developed anaphase aberration test that utilizes sea urchin embryos. Other related efforts include field validation studies of the new tests, evaluationmore » of their potential ecological relevance, and analysis of their sensitivity relative to that of existing toxicity tests that assess only lethal effects, rather than genetic damage.« less

  1. Urine assay for tenofovir to monitor adherence in real time to tenofovir disoproxil fumarate/emtricitabine as pre-exposure prophylaxis.

    PubMed

    Koenig, H C; Mounzer, K; Daughtridge, G W; Sloan, C E; Lalley-Chareczko, L; Moorthy, G S; Conyngham, S C; Zuppa, A F; Montaner, L J; Tebas, P

    2017-07-01

    Tenofovir disoproxil fumarate/emtricitabine (TDF/FTC) is approved for pre-exposure prophylaxis (PrEP) against HIV infection. Adherence is critical for the success of PrEP, but current adherence measurements are inadequate for real-time adherence monitoring. We developed and validated a urine assay to measure tenofovir (TFV) to objectively monitor adherence to PrEP. We developed a urine assay using high-performance liquid chromatography coupled to tandem mass spectrometry with high sensitivity/specificity for TFV that allowed us to determine TFV concentrations in log 10 categories between 0 and 10 000 ng/mL. We validated the assay in three cohorts: (1) HIV-positive subjects with undetectable viral loads on a TDF/FTC-based regimen, (2) healthy HIV-negative subjects who received a single dose of TDF/FTC, and (3) HIV-negative subjects receiving daily TDF/FTC as PrEP for 24 weeks. The urine assay detected TFV with greater sensitivity than plasma-based measures and with a window of measurements within 7 days of the last TDF/FTC dose. Based on the urine log-linear clearance after the last dose and its concordance with all detectable plasma levels, a urine TFV concentration > 1000 ng/mL was identified as highly predictive of the presence of TFV in plasma at > 10 ng/mL. The urine assay was able to distinguish high and low adherence patterns within the last 48 h (> 1000 ng/mL versus 10-1000 ng/mL), as well as nonadherence (< 10 ng/mL) extended over at least 1 week prior to measurement. We provide proof of concept that a semiquantitative urine assay measuring levels of TFV could be further developed into a point-of-care test and be a useful tool to monitor adherence to PrEP. © 2017 British HIV Association.

  2. Facile Arsenazo III-Based Assay for Monitoring Rare Earth Element Depletion from Cultivation Media for Methanotrophic and Methylotrophic Bacteria

    PubMed Central

    Hogendoorn, Carmen; Roszczenko-Jasińska, Paula; Martinez-Gomez, N. Cecilia; de Graaff, Johann; Grassl, Patrick; Pol, Arjan; Op den Camp, Huub J. M.

    2018-01-01

    ABSTRACT Recently, methanotrophic and methylotrophic bacteria were found to utilize rare earth elements (REEs). To monitor the REE content in culture media of these bacteria, we have developed a rapid screening method using the Arsenazo III (AS III) dye for spectrophotometric REE detection in the low μM (0.1 to 10 μM) range. We designed this assay to follow LaIII and EuIII depletion from the culture medium by the acidophilic verrucomicrobial methanotroph Methylacidiphilum fumariolicum strain SolV. The assay can also be modified to screen the uptake of other REEs, such as PrIII, or to monitor the depletion of LaIII from growth media in neutrophilic methylotrophs such as Methylobacterium extorquens strain AM1. The AS III assay presents a convenient and fast detection method for REE levels in culture media and is a sensitive alternative to inductively coupled plasma mass spectrometry (ICP-MS) or atomic absorption spectroscopy (AAS). IMPORTANCE REE-dependent bacterial metabolism is a quickly emerging field, and while the importance of REEs for both methanotrophic and methylotrophic bacteria is now firmly established, many important questions, such as how these insoluble elements are taken up into cells, are still unanswered. Here, an Arsenazo III dye-based assay has been developed for fast, specific, and sensitive determination of REE content in different culture media. This assay presents a useful tool for optimizing cultivation protocols, as well as for routine REE monitoring during bacterial growth without the need for specialized analytical instrumentation. Furthermore, this assay has the potential to promote the discovery of other REE-dependent microorganisms and can help to elucidate the mechanisms for acquisition of REEs by methanotrophic and methylotrophic bacteria. PMID:29453257

  3. Facile Arsenazo III-Based Assay for Monitoring Rare Earth Element Depletion from Cultivation Media for Methanotrophic and Methylotrophic Bacteria.

    PubMed

    Hogendoorn, Carmen; Roszczenko-Jasińska, Paula; Martinez-Gomez, N Cecilia; de Graaff, Johann; Grassl, Patrick; Pol, Arjan; Op den Camp, Huub J M; Daumann, Lena J

    2018-04-15

    Recently, methanotrophic and methylotrophic bacteria were found to utilize rare earth elements (REEs). To monitor the REE content in culture media of these bacteria, we have developed a rapid screening method using the Arsenazo III (AS III) dye for spectrophotometric REE detection in the low μM (0.1 to 10 μM) range. We designed this assay to follow La III and Eu III depletion from the culture medium by the acidophilic verrucomicrobial methanotroph Methylacidiphilum fumariolicum strain SolV. The assay can also be modified to screen the uptake of other REEs, such as Pr III , or to monitor the depletion of La III from growth media in neutrophilic methylotrophs such as Methylobacterium extorquens strain AM1. The AS III assay presents a convenient and fast detection method for REE levels in culture media and is a sensitive alternative to inductively coupled plasma mass spectrometry (ICP-MS) or atomic absorption spectroscopy (AAS). IMPORTANCE REE-dependent bacterial metabolism is a quickly emerging field, and while the importance of REEs for both methanotrophic and methylotrophic bacteria is now firmly established, many important questions, such as how these insoluble elements are taken up into cells, are still unanswered. Here, an Arsenazo III dye-based assay has been developed for fast, specific, and sensitive determination of REE content in different culture media. This assay presents a useful tool for optimizing cultivation protocols, as well as for routine REE monitoring during bacterial growth without the need for specialized analytical instrumentation. Furthermore, this assay has the potential to promote the discovery of other REE-dependent microorganisms and can help to elucidate the mechanisms for acquisition of REEs by methanotrophic and methylotrophic bacteria. Copyright © 2018 Hogendoorn et al.

  4. 40 CFR 60.2972 - How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... curtain incinerators that burn only wood waste, clean lumber, and yard waste? 60.2972 Section 60.2972... Commenced on or After June 16, 2006 Air Curtain Incinerators That Burn Only Wood Waste, Clean Lumber, and Yard Waste § 60.2972 How must I monitor opacity for air curtain incinerators that burn only wood waste...

  5. 40 CFR 60.2972 - How must I monitor opacity for air curtain incinerators that burn only wood waste, clean lumber...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... curtain incinerators that burn only wood waste, clean lumber, and yard waste? 60.2972 Section 60.2972... Commenced on or After June 16, 2006 Air Curtain Incinerators That Burn Only Wood Waste, Clean Lumber, and Yard Waste § 60.2972 How must I monitor opacity for air curtain incinerators that burn only wood waste...

  6. A microfluidic platform with integrated arrays for immunologic assays for biological pathogen detection

    NASA Astrophysics Data System (ADS)

    Klemm, Richard; Becker, Holger; Hlawatsch, Nadine; Julich, Sandra; Miethe, Peter; Moche, Christian; Schattschneider, Sebastian; Tomaso, Herbert; Gärtner, Claudia

    2014-05-01

    The ability to integrate complete assays on a microfluidic chip helps to greatly simplify instrument requirements and allows the use of lab-on-a-chip technology in the field. A core application for such field-portable systems is the detection of pathogens in a CBRN scenario such as permanent monitoring of airborne pathogens, e.g. in subway stations or hospitals etc. An immunological assay was chosen as method for the pathogen identification. The conceptual approach was its realization as a lab-on-a-chip system, enabling an easy handling of the sample in an automated manner. The immunological detection takes place on an antibody array directly implemented in the microfluidic network. Different immobilization strategies will be presented showing the performance of the system. Central elements of the disposable microfluidic device like fluidic interface, turning valves, liquid introduction and waste storage, as well as the architecture of measurement and control fluidic network, will be introduced. Overall process times of about 30 minutes were achieved and assays for the detection of Francisella tularensis and Yersinia pestis are presented. An important feature of the integrated lab-on-a-chip approach is that all waste liquids remain on-chip and contamination risks can be avoided.

  7. Monitoring of olive oil mills' wastes using electrical resistivity tomography techniques

    NASA Astrophysics Data System (ADS)

    Simyrdanis, Kleanthis; Papadopoulos, Nikos; Kirkou, Stella; Sarris, Apostolos; Tsourlos, Panagiotis

    2014-08-01

    Olive oil mills' wastes (OOMW) are one of the byproducts of the oil production that can lead to serious environmental pollution when they are deposited in ponds dug on the ground surface. Electrical Resistivity Tomography (ERT) method can provide a valuable tool in order to monitor through time the physical flow of the wastes into the subsurface. ERT could potentially locate the electrical signature due to lower resistivity values resulting from the leakage of OOMW to the subsurface. For this purpose, two vertical boreholes were installed (12m depth, 9 m apart) in the vicinity of an existing pond which is filled with OOMW during the oil production period. The test site is situated in Saint Andreas village about 15km south of the city of Rethymno (Crete, Greece). Surface ERT measurements were collected along multiple lines in order to reconstruct the subsurface resistivity models. Data acquisition was performed with standard and optimized electrode configuration protocols. The monitoring survey includes the ERT data collection for a period of time. The study was initiated before the OOMW were deposited in the pond, so resistivity fluctuations are expected due to the flow of OOMW in the porous subsurface media through time. Preliminary results show the good correlation of the ERT images with the drilled geological formations and the identification of low resistivity subsurface zone that could be attributed to the flow of the wastes within the porous layers.

  8. 40 CFR Table 7 to Subpart Ggg of... - Wastewater-Inspection and Monitoring Requirements for Waste Management Units

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 12 2012-07-01 2011-07-01 true Wastewater-Inspection and Monitoring..., Subpt. GGG, Table 7 Table 7 to Subpart GGG of Part 63—Wastewater—Inspection and Monitoring Requirements for Waste Management Units To comply with Inspection or monitoring requirement Frequency of inspection...

  9. 40 CFR Table 7 to Subpart Ggg of... - Wastewater-Inspection and Monitoring Requirements for Waste Management Units

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 11 2011-07-01 2011-07-01 false Wastewater-Inspection and Monitoring..., Subpt. GGG, Table 7 Table 7 to Subpart GGG of Part 63—Wastewater—Inspection and Monitoring Requirements for Waste Management Units To comply with Inspection or monitoring requirement Frequency of inspection...

  10. The monitoring of organic waste pollution in the sibelis river

    NASA Astrophysics Data System (ADS)

    Huda, Thorikul; Jannah, Wirdatul

    2017-03-01

    Has conducted monitoring of organic waste pollution in the River Sibelis of Tegal City of Central Java. Organic wastes that pollute River Sibelis can degrade the quality of well water along the river. Monitoring carried out in the upstream and downstream by chemical oxygen demand (COD) and biochemical oxygen demand (BOD) parameters. COD test methods by titration and the results are used to determine the test sample comparison with the volume of diluent required for analysts BOD. COD test results on the upstream and downstream Sibelis River respectively 58.13 mg/L and 73.97 mg / L so that the ratio of the test sample with diluent volume for BOD analysis is 20: 280 (Sawyer, 1978). BOD test principle is based on the reduction of dissolved oxygen zero day (DO0) and five days (DO5). The result of observation BOD samples at upstream and downstream Sibelis Rivers are 10.7212 mg / L and 5.3792 mg / L respectively. Quality control of BOD testing conducted with measurement accuracy and precision and obtained result are 85.36% and 0.27% respectively. The result of uncertainty measurement for BOD testing at upstream and downstream are ±0.4469 mg/L and ±0.22188 mg/L.

  11. Performance of the New Bayer VERSANT HCV RNA 3.0 assay for quantitation of hepatitis C virus RNA in plasma and serum: conversion to international units and comparison with the Roche COBAS Amplicor HCV Monitor, Version 2.0, assay.

    PubMed

    Beld, Marcel; Sentjens, Roel; Rebers, Sjoerd; Weegink, Christine; Weel, Jan; Sol, Cees; Boom, René

    2002-03-01

    We have evaluated the VERSANT HCV RNA 3.0. Assay (HCV 3.0 bDNA assay) (Bayer Diagnostics, Berkeley, Calif.), which is an improved signal amplification procedure for the HCV 2.0 bDNA assay for the quantitation of hepatitis C virus (HCV) RNA in serum or plasma of HCV-infected individuals. The HCV 3.0 bDNA assay has a linear dynamic range of 2.5 x 10(3) to 4.0 x 10(7) HCV RNA copies per ml (c/ml). The performance of the HCV 3.0 bDNA assay was evaluated using three different test panels. An overall specificity of 96.8% relative to the detection limit of the HCV 3.0 bDNA assay was found. The intra- and interrun reproducibilities for both the dilution panel and the NAP (AcroMetrix, Benicia, Calif.) panel were consistent with coefficients of variation of less than 9%. Quantitation with the HCV 3.0 bDNA assay was linear over the entire range of both panels (ranges of 4.4 x 10(3) to 3.5 x 10(6) c/ml and 5 x 10(3) to 2 x 10(6) IU/ml, respectively), with correlation coefficients of 0.999, slopes close to one, and intercepts close to zero. The regression equation indicated that 1 IU corresponded to about 4.8 copies of HCV RNA. A correlation coefficient of 0.941 was found for HCV RNA values (in international units per milliliter) obtained from the HCV 3.0 bDNA assay and the HCV Monitor version 2.0 assay (HCV Monitor 2.0 assay) (Roche Diagnostic Systems, Branchburg, N.J.). Quantitative results obtained close to the lower limit of the HCV 3.0 bDNA assay might imply that its lower limit should be reconsidered and raised, if necessary. It appeared that quantitation values obtained from the HCV Monitor 2.0 assay of between 5 x 10(2) and 10(5) IU/ml were in general higher than those obtained from the HCV 3.0 bDNA assay, whereas values obtained from the HCV Monitor 2.0 assay were underestimated for samples with HCV RNA levels above 10(5) IU/ml.

  12. Performance of the New Bayer VERSANT HCV RNA 3.0 Assay for Quantitation of Hepatitis C Virus RNA in Plasma and Serum: Conversion to International Units and Comparison with the Roche COBAS Amplicor HCV Monitor, Version 2.0, Assay

    PubMed Central

    Beld, Marcel; Sentjens, Roel; Rebers, Sjoerd; Weegink, Christine; Weel, Jan; Sol, Cees; Boom, René

    2002-01-01

    We have evaluated the VERSANT HCV RNA 3.0. Assay (HCV 3.0 bDNA assay) (Bayer Diagnostics, Berkeley, Calif.), which is an improved signal amplification procedure for the HCV 2.0 bDNA assay for the quantitation of hepatitis C virus (HCV) RNA in serum or plasma of HCV-infected individuals. The HCV 3.0 bDNA assay has a linear dynamic range of 2.5 × 103 to 4.0 × 107 HCV RNA copies per ml (c/ml). The performance of the HCV 3.0 bDNA assay was evaluated using three different test panels. An overall specificity of 96.8% relative to the detection limit of the HCV 3.0 bDNA assay was found. The intra- and interrun reproducibilities for both the dilution panel and the NAP (AcroMetrix, Benicia, Calif.) panel were consistent with coefficients of variation of less than 9%. Quantitation with the HCV 3.0 bDNA assay was linear over the entire range of both panels (ranges of 4.4 × 103 to 3.5 × 106 c/ml and 5 × 103 to 2 × 106 IU/ml, respectively), with correlation coefficients of 0.999, slopes close to one, and intercepts close to zero. The regression equation indicated that 1 IU corresponded to about 4.8 copies of HCV RNA. A correlation coefficient of 0.941 was found for HCV RNA values (in international units per milliliter) obtained from the HCV 3.0 bDNA assay and the HCV Monitor version 2.0 assay (HCV Monitor 2.0 assay) (Roche Diagnostic Systems, Branchburg, N.J.). Quantitative results obtained close to the lower limit of the HCV 3.0 bDNA assay might imply that its lower limit should be reconsidered and raised, if necessary. It appeared that quantitation values obtained from the HCV Monitor 2.0 assay of between 5 × 102 and 105 IU/ml were in general higher than those obtained from the HCV 3.0 bDNA assay, whereas values obtained from the HCV Monitor 2.0 assay were underestimated for samples with HCV RNA levels above 105 IU/ml. PMID:11880394

  13. 40 CFR Table 7 to Subpart Ggg of... - Wastewater-Inspection and Monitoring Requirements for Waste Management Units

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 12 2014-07-01 2014-07-01 false Wastewater-Inspection and Monitoring... Production Pt. 63, Subpt. GGG, Table 7 Table 7 to Subpart GGG of Part 63—Wastewater—Inspection and Monitoring Requirements for Waste Management Units To comply with Inspection or monitoring requirement Frequency of...

  14. 40 CFR Table 7 to Subpart Ggg of... - Wastewater-Inspection and Monitoring Requirements for Waste Management Units

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 12 2013-07-01 2013-07-01 false Wastewater-Inspection and Monitoring... Production Pt. 63, Subpt. GGG, Table 7 Table 7 to Subpart GGG of Part 63—Wastewater—Inspection and Monitoring Requirements for Waste Management Units To comply with Inspection or monitoring requirement Frequency of...

  15. A Multiplexed Assay That Monitors Effects of Multiple Compound Treatment Times Reveals Candidate Immune-Enhancing Compounds.

    PubMed

    Zhao, Ziyan; Henowitz, Liza; Zweifach, Adam

    2018-05-01

    We previously developed a flow cytometry assay that monitored lytic granule exocytosis in cytotoxic T lymphocytes stimulated by contacting beads coated with activating anti-CD3 antibodies. That assay was multiplexed in that responses of cells that did or did not receive the activating stimulus were distinguished via changes in light scatter accompanying binding of cells to beads, allowing us to discriminate compounds that activate responses on their own from compounds that enhance responses in cells that received the activating stimulus, all within a single sample. Here we add a second dimension of multiplexing by developing means to assess in a single sample the effects of treating cells with test compounds for different times. Bar-coding cells before adding them to test wells lets us determine compound treatment time while also monitoring activation status and response amplitude at the point of interrogation. This multiplexed assay is suitable for screening 96-well plates. We used it to screen compounds from the National Cancer Institute, identifying several compounds that enhance anti-LAMP1 responses. Multiple-treatment-time (MTT) screening enabled by bar-coding and read via high-throughput flow cytometry may be a generally useful method for facilitating the discovery of compounds of interest.

  16. 40 CFR 62.14820 - How must I monitor opacity for air curtain incinerators that burn 100 percent wood wastes, clean...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... curtain incinerators that burn 100 percent wood wastes, clean lumber, and/or yard waste? 62.14820 Section... Before November 30, 1999 Air Curtain Incinerators That Burn 100 Percent Wood Wastes, Clean Lumber And/or Yard Waste § 62.14820 How must I monitor opacity for air curtain incinerators that burn 100 percent...

  17. 40 CFR 62.14820 - How must I monitor opacity for air curtain incinerators that burn 100 percent wood wastes, clean...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... curtain incinerators that burn 100 percent wood wastes, clean lumber, and/or yard waste? 62.14820 Section... Before November 30, 1999 Air Curtain Incinerators That Burn 100 Percent Wood Wastes, Clean Lumber And/or Yard Waste § 62.14820 How must I monitor opacity for air curtain incinerators that burn 100 percent...

  18. 40 CFR 62.14820 - How must I monitor opacity for air curtain incinerators that burn 100 percent wood wastes, clean...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... curtain incinerators that burn 100 percent wood wastes, clean lumber, and/or yard waste? 62.14820 Section... Before November 30, 1999 Air Curtain Incinerators That Burn 100 Percent Wood Wastes, Clean Lumber And/or Yard Waste § 62.14820 How must I monitor opacity for air curtain incinerators that burn 100 percent...

  19. Multiple reaction monitoring assay based on conventional liquid chromatography and electrospray ionization for simultaneous monitoring of multiple cerebrospinal fluid biomarker candidates for Alzheimer's disease

    PubMed Central

    Choi1, Yong Seok; Lee, Kelvin H.

    2016-01-01

    Alzheimer's disease (AD) is the most common type of dementia, but early and accurate diagnosis remains challenging. Previously, a panel of cerebrospinal fluid (CSF) biomarker candidates distinguishing AD and non-AD CSF accurately (> 90%) was reported. Furthermore, a multiple reaction monitoring (MRM) assay based on nano liquid chromatography tandem mass spectrometry (nLC-MS/MS) was developed to help validate putative AD CSF biomarker candidates including proteins from the panel. Despite the good performance of the MRM assay, wide acceptance may be challenging because of limited availability of nLC-MS/MS systems laboratories. Thus, here, a new MRM assay based on conventional LC-MS/MS is presented. This method monitors 16 peptides representing 16 (of 23) biomarker candidates that belonged to the previous AD CSF panel. A 30-times more concentrated sample than the sample used for the previous study was loaded onto a high capacity trap column, and all 16 MRM transitions showed good linearity (average R2 = 0.966), intra-day reproducibility (average coefficient of variance (CV) = 4.78%), and inter-day reproducibility (average CV = 9.85%). The present method has several advantages such as a shorter analysis time, no possibility of target variability, and no need for an internal standard. PMID:26404792

  20. Multiple reaction monitoring assay based on conventional liquid chromatography and electrospray ionization for simultaneous monitoring of multiple cerebrospinal fluid biomarker candidates for Alzheimer's disease.

    PubMed

    Choi, Yong Seok; Lee, Kelvin H

    2016-03-01

    Alzheimer's disease (AD) is the most common type of dementia, but early and accurate diagnosis remains challenging. Previously, a panel of cerebrospinal fluid (CSF) biomarker candidates distinguishing AD and non-AD CSF accurately (>90 %) was reported. Furthermore, a multiple reaction monitoring (MRM) assay based on nano liquid chromatography tandem mass spectrometry (nLC-MS/MS) was developed to help validate putative AD CSF biomarker candidates including proteins from the panel. Despite the good performance of the MRM assay, wide acceptance may be challenging because of limited availability of nLC-MS/MS systems in laboratories. Thus, here, a new MRM assay based on conventional LC-MS/MS is presented. This method monitors 16 peptides representing 16 (of 23) biomarker candidates that belonged to the previous AD CSF panel. A 30-times more concentrated sample than the sample used for the previous study was loaded onto a high capacity trap column, and all 16 MRM transitions showed good linearity (average R(2) = 0.966), intra-day reproducibility (average coefficient of variance (CV) = 4.78 %), and inter-day reproducibility (average CV = 9.85 %). The present method has several advantages such as a shorter analysis time, no possibility of target variability, and no need for an internal standard.

  1. Genetic Assay for Transcription Errors: Methods to Monitor Treatments or Chemicals that Increase the Error Rate of RNA synthesis | NCI Technology Transfer Center | TTC

    Cancer.gov

    Researchers at the National Cancer Institute (NCI) developed a genetic assay for detecting transcription errors in RNA synthesis. This new assay extends the familiar concept of an Ames test which monitors DNA damage and synthesis errors to the previously inaccessible issue of RNA synthesis fidelity. The FDA requires genetic DNA focused tests for all drug approval as it assesses the in vivo mutagenic and carcinogenic potential of a drug. The new assay will open an approach to monitoring the impact of treatments on the accuracy of RNA synthesis. Errors in transcription have been hypothesized to be a component of aging and age-related diseases. The National Cancer Institute (NCI) seeks licensing partners for the genetic assay.

  2. Neutron coincidence counting based on time interval analysis with one- and two-dimensional Rossi-alpha distributions: an application for passive neutron waste assay

    NASA Astrophysics Data System (ADS)

    Bruggeman, M.; Baeten, P.; De Boeck, W.; Carchon, R.

    1996-02-01

    Neutron coincidence counting is commonly used for the non-destructive assay of plutonium bearing waste or for safeguards verification measurements. A major drawback of conventional coincidence counting is related to the fact that a valid calibration is needed to convert a neutron coincidence count rate to a 240Pu equivalent mass ( 240Pu eq). In waste assay, calibrations are made for representative waste matrices and source distributions. The actual waste however may have quite different matrices and source distributions compared to the calibration samples. This often results in a bias of the assay result. This paper presents a new neutron multiplicity sensitive coincidence counting technique including an auto-calibration of the neutron detection efficiency. The coincidence counting principle is based on the recording of one- and two-dimensional Rossi-alpha distributions triggered respectively by pulse pairs and by pulse triplets. Rossi-alpha distributions allow an easy discrimination between real and accidental coincidences and are aimed at being measured by a PC-based fast time interval analyser. The Rossi-alpha distributions can be easily expressed in terms of a limited number of factorial moments of the neutron multiplicity distributions. The presented technique allows an unbiased measurement of the 240Pu eq mass. The presented theory—which will be indicated as Time Interval Analysis (TIA)—is complementary to Time Correlation Analysis (TCA) theories which were developed in the past, but is from the theoretical point of view much simpler and allows a straightforward calculation of deadtime corrections and error propagation. Analytical expressions are derived for the Rossi-alpha distributions as a function of the factorial moments of the efficiency dependent multiplicity distributions. The validity of the proposed theory is demonstrated and verified via Monte Carlo simulations of pulse trains and the subsequent analysis of the simulated data.

  3. 40 CFR 60.1450 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... curtain incinerators that burn 100 percent yard waste? 60.1450 Section 60.1450 Protection of Environment... Air Curtain Incinerators That Burn 100 Percent Yard Waste § 60.1450 How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste? (a) Use EPA Reference Method 9 in appendix A of...

  4. 40 CFR 60.1450 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... curtain incinerators that burn 100 percent yard waste? 60.1450 Section 60.1450 Protection of Environment... Air Curtain Incinerators That Burn 100 Percent Yard Waste § 60.1450 How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste? (a) Use EPA Reference Method 9 in appendix A of...

  5. 40 CFR 60.1450 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... curtain incinerators that burn 100 percent yard waste? 60.1450 Section 60.1450 Protection of Environment... Air Curtain Incinerators That Burn 100 Percent Yard Waste § 60.1450 How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste? (a) Use EPA Reference Method 9 in appendix A of...

  6. 40 CFR 60.1925 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... curtain incinerators that burn 100 percent yard waste? 60.1925 Section 60.1925 Protection of Environment... or Before August 30, 1999 Model Rule-Air Curtain Incinerators That Burn 100 Percent Yard Waste § 60.1925 How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste? (a) Use...

  7. 40 CFR 60.1925 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... curtain incinerators that burn 100 percent yard waste? 60.1925 Section 60.1925 Protection of Environment... or Before August 30, 1999 Model Rule-Air Curtain Incinerators That Burn 100 Percent Yard Waste § 60.1925 How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste? (a) Use...

  8. 40 CFR 60.1925 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... curtain incinerators that burn 100 percent yard waste? 60.1925 Section 60.1925 Protection of Environment... or Before August 30, 1999 Model Rule-Air Curtain Incinerators That Burn 100 Percent Yard Waste § 60.1925 How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste? (a) Use...

  9. 40 CFR 60.1925 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... curtain incinerators that burn 100 percent yard waste? 60.1925 Section 60.1925 Protection of Environment... or Before August 30, 1999 Model Rule-Air Curtain Incinerators That Burn 100 Percent Yard Waste § 60.1925 How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste? (a) Use...

  10. 40 CFR 60.1925 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... curtain incinerators that burn 100 percent yard waste? 60.1925 Section 60.1925 Protection of Environment... or Before August 30, 1999 Model Rule-Air Curtain Incinerators That Burn 100 Percent Yard Waste § 60.1925 How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste? (a) Use...

  11. 40 CFR 60.1450 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... curtain incinerators that burn 100 percent yard waste? 60.1450 Section 60.1450 Protection of Environment... Air Curtain Incinerators That Burn 100 Percent Yard Waste § 60.1450 How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste? (a) Use EPA Reference Method 9 in appendix A of...

  12. 40 CFR 60.1450 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... curtain incinerators that burn 100 percent yard waste? 60.1450 Section 60.1450 Protection of Environment... Air Curtain Incinerators That Burn 100 Percent Yard Waste § 60.1450 How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste? (a) Use EPA Reference Method 9 in appendix A of...

  13. Hydrologic monitoring of a waste-injection well near Milton, Florida, June 1975 - June 1977

    USGS Publications Warehouse

    Pascale, Charles A.; Martin, J.B.

    1978-01-01

    This report presents the hydraulic and chemical data collected from June 1, 1975, when injection began, to June 30, 1977 through a monitoring program at a deep-well waste-injection system at the American Cyanamid Company's plant near Milton, about 12 miles northwest of Pensacola. The injection system consists of a primary injection well, a standby injection well, and two deep monitor wells all completed open hole in the lower limestone of the Floridan aquifer and one shallow-monitor well completed in the upper limestone of the Floridan aquifer. Two of the monitor wells and the standby injection well are used to observe hydraulic and geochemical effects of waste injection in the injection zone at locations 8,180 feet northeast, 1,560 feet south, and 1,025 feet southwest of the primary injection well. The shallow-monitor well, used to observe any effects in the first permeable zone above the 200-foot-thick confining bed, is 28 feet north of the primary injection well. Since injection began in June 1975, 607 million gallons of treated industrial liquid waste with a pH of 4.6 to 6.3 and containing high concentrations of nitrate, organic nitrogen and carbon have been injected into a saline-water-filled limestone aquifer. Wellhead pressure at the injection well in June 1977 average 137 pounds per square inch and the hydraulic pressure gradient was 0.53 pound per square inch per foot of depth to the top of the injection zone. Water levels rose from 36 to 74 feet at the three wells used to monitor the injection zone during the 25-month period. The water level in the shallow-monitor well declined about 8 feet. No changes were detected in the chemical character of water from the shallow-monitor well and deep-monitor well-north. Increases in concentration of bicarbonate and dissolved organic carbon were detected in water from the deep-test monitor well in February 1976 and at the standby injection well in August 1976. In addition to increases in bicarbonate and dissolved

  14. Developing a monitoring and evaluation framework to integrate and formalize the informal waste and recycling sector: the case of the Philippine National Framework Plan.

    PubMed

    Serrona, Kevin Roy B; Yu, Jeongsoo; Aguinaldo, Emelita; Florece, Leonardo M

    2014-09-01

    The Philippines has been making inroads in solid waste management with the enactment and implementation of the Republic Act 9003 or the Ecological Waste Management Act of 2000. Said legislation has had tremendous influence in terms of how the national and local government units confront the challenges of waste management in urban and rural areas using the reduce, reuse, recycle and recovery framework or 4Rs. One of the sectors needing assistance is the informal waste sector whose aspiration is legal recognition of their rank and integration of their waste recovery activities in mainstream waste management. To realize this, the Philippine National Solid Waste Management Commission initiated the formulation of the National Framework Plan for the Informal Waste Sector, which stipulates approaches, strategies and methodologies to concretely involve the said sector in different spheres of local waste management, such as collection, recycling and disposal. What needs to be fleshed out is the monitoring and evaluation component in order to gauge qualitative and quantitative achievements vis-a-vis the Framework Plan. In the process of providing an enabling environment for the informal waste sector, progress has to be monitored and verified qualitatively and quantitatively and measured against activities, outputs, objectives and goals. Using the Framework Plan as the reference, this article developed monitoring and evaluation indicators using the logical framework approach in project management. The primary objective is to institutionalize monitoring and evaluation, not just in informal waste sector plans, but in any waste management initiatives to ensure that envisaged goals are achieved. © The Author(s) 2014.

  15. Assays for therapeutic drug monitoring of β-lactam antibiotics: A structured review.

    PubMed

    Carlier, Mieke; Stove, Veronique; Wallis, Steven C; De Waele, Jan J; Verstraete, Alain G; Lipman, Jeffrey; Roberts, Jason A

    2015-10-01

    In some patient groups, including critically ill patients, the pharmacokinetics of β-lactam antibiotics may be profoundly disturbed due to pathophysiological changes in distribution and elimination. Therapeutic drug monitoring (TDM) is a strategy that may help to optimise dosing. The aim of this review was to identify and analyse the published literature on the methods used for β-lactam quantification in TDM programmes. Sixteen reports described methods for the simultaneous determination of three or more β-lactam antibiotics in plasma/serum. Measurement of these antibiotics, due to low frequency of usage relative to some other tests, is generally limited to in-house chromatographic methods coupled to ultraviolet or mass spectrometric detection. Although many published methods state they are fit for TDM, they are inconvenient because of intensive sample preparation and/or long run times. Ideally, methods used for routine TDM should have a short turnaround time (fast run-time and fast sample preparation), a low limit of quantification and a sufficiently high upper limit of quantification. The published assays included a median of 6 analytes [interquartile range (IQR) 4-10], with meropenem and piperacillin being the most frequently measured β-lactam antibiotics. The median run time was 8 min (IQR 5.9-21.3 min). There is also a growing number of methods measuring free concentrations. An assay that measures antibiotics without any sample preparation would be the next step towards real-time monitoring; no such method is currently available. Copyright © 2015 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  16. Real-time monitoring of enzyme-free strand displacement cascades by colorimetric assays

    NASA Astrophysics Data System (ADS)

    Duan, Ruixue; Wang, Boya; Hong, Fan; Zhang, Tianchi; Jia, Yongmei; Huang, Jiayu; Hakeem, Abdul; Liu, Nannan; Lou, Xiaoding; Xia, Fan

    2015-03-01

    The enzyme-free toehold-mediated strand displacement reaction has shown potential for building programmable DNA circuits, biosensors, molecular machines and chemical reaction networks. Here we report a simple colorimetric method using gold nanoparticles as signal generators for the real-time detection of the product of the strand displacement cascade. During the process the assembled gold nanoparticles can be separated, resulting in a color change of the solution. This assay can also be applied in complex mixtures, fetal bovine serum, and to detect single-base mismatches. These results suggest that this method could be of general utility to monitor more complex enzyme-free strand displacement reaction-based programmable systems or for further low-cost diagnostic applications.The enzyme-free toehold-mediated strand displacement reaction has shown potential for building programmable DNA circuits, biosensors, molecular machines and chemical reaction networks. Here we report a simple colorimetric method using gold nanoparticles as signal generators for the real-time detection of the product of the strand displacement cascade. During the process the assembled gold nanoparticles can be separated, resulting in a color change of the solution. This assay can also be applied in complex mixtures, fetal bovine serum, and to detect single-base mismatches. These results suggest that this method could be of general utility to monitor more complex enzyme-free strand displacement reaction-based programmable systems or for further low-cost diagnostic applications. Electronic supplementary information (ESI) available: Experimental procedures and analytical data are provided. See DOI: 10.1039/c5nr00697j

  17. Clinical performance of the LCx HCV RNA quantitative assay.

    PubMed

    Bertuzis, Rasa; Hardie, Alison; Hottentraeger, Barbara; Izopet, Jacques; Jilg, Wolfgang; Kaesdorf, Barbara; Leckie, Gregor; Leete, Jean; Perrin, Luc; Qiu, Chunfu; Ran, Iris; Schneider, George; Simmonds, Peter; Robinson, John

    2005-02-01

    This study was conducted to assess the performance of the Abbott laboratories LCx HCV RNA Quantitative Assay (LCx assay) in the clinical setting. Four clinical laboratories measured LCx assay precision, specificity, and linearity. In addition, a method comparison was conducted between the LCx assay and the Roche HCV Amplicor Monitor, version 2.0 (Roche Monitor 2.0) and the Bayer VERSANT HCV RNA 3.0 Assay (Bayer bDNA 3.0) quantitative assays. For precision, the observed LCx assay intra-assay standard deviation (S.D.) was 0.060-0.117 log IU/ml, the inter-assay S.D. was 0.083-0.133 log IU/ml, the inter-lot S.D. was 0.105-0.177 log IU/ml, the inter-site S.D. was 0.099-0.190 log IU/ml, and the total S.D. was 0.113-0.190 log IU/ml. The specificity of the LCx assay was 99.4% (542/545; 95% CI, 98.4-99.9%). For linearity, the mean pooled LCx assay results were linear (r=0.994) over the range of the panel (2.54-5.15 log IU/ml). A method comparison demonstrated a correlation coefficient of 0.881 between the LCx assay and Roche Monitor 2.0, 0.872 between the LCx assay and Bayer bDNA 3.0, and 0.870 between Roche Monitor 2.0 and Bayer bDNA 3.0. The mean LCx assay result was 0.04 log IU/ml (95% CI, -0.08, 0.01) lower than the mean Roche Monitor 2.0 result, but 0.57 log IU/ml (95% CI, 0.53, 0.61) higher than the mean Bayer bDNA 3.0 result. The mean Roche Monitor 2.0 result was 0.60 log IU/ml (95% CI, 0.56, 0.65) higher than the mean Bayer bDNA 3.0 result. The LCx assay quantitated genotypes 1-4 with statistical equivalency. The vast majority (98.9%, 278/281) of paired LCx assay-Roche Monitor 2.0 specimen results were within 1 log IU/ml. Similarly, 86.6% (240/277) of paired LCx assay and Bayer bDNA 3.0 specimen results were within 1 log, as were 85.6% (237/277) of paired Roche Monitor 2.0 and Bayer specimen results. These data demonstrate that the LCx assay may be used for quantitation of HCV RNA in HCV-infected individuals.

  18. Monitoring the Durability Performance of Concrete in Nuclear Waste Containment. Technical Progress Report No. 3

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ulm, Franz-Josef

    2000-03-31

    OAK-B135 Monitoring the Durability Performance of Concrete in Nuclear Waste Containment. Technical Progress Report No. 3(NOTE: Part II A item 1 indicates ''PAPER'', but a report is attached electronically)

  19. Monitoring cyclodextrin-polyviologen pseudopolyrotaxanes with the Bradford assay.

    PubMed

    Belitsky, Jason M; Nelson, Alshakim; Stoddart, J Fraser

    2006-01-21

    Self-assembled multivalent pseudopolyrotaxanes, composed of lactoside-bearing cyclodextrin (CD) rings threaded on linear polyviologen polymers, have been introduced recently as flexible and dynamic neoglycoconjugates. In the course of this research, it was found that polyviologens are responsive to the Bradford assay, which is traditionally highly selective for proteins. The response of the pseudopolyrotaxanes to the Bradford assay was dependant on, and thus indicative of, the degree of threading of the CD rings onto the polyelectrolyte. The assay was then used to report on the threading and dethreading of native and lactoside-bearing alpha-CD rings onto and off of polyviologen chains, a phenomenon which demonstrates the utility of biochemical assays to address problems unique to supramolecular chemistry.

  20. Can HIV reverse transcriptase activity assay be a low-cost alternative for viral load monitoring in resource-limited settings?

    PubMed

    Gupta, Soham; Palchaudhuri, Riya; Neogi, Ujjwal; Srinivasa, Hiresave; Ashorn, Per; De Costa, Ayesha; Källander, Clas; Shet, Anita

    2016-01-27

    To evaluate the performance and cost of an HIV reverse transcriptase-enzyme activity (HIV-RT) assay in comparison to an HIV-1 RNA assay for routine viral load monitoring in resource limited settings. A cohort-based longitudinal study. Two antiretroviral therapy (ART) centres in Karnataka state, South India, providing treatment under the Indian AIDS control programme. A cohort of 327 HIV-1-infected Indian adult patients initiating first-line ART. Performance and cost of an HIV-RT assay (ExaVir Load V3) in comparison to a gold standard HIV-1 RNA assay (Abbott m2000rt) in a cohort of 327 Indian patients before (WK00) and 4 weeks (WK04) after initiation of first-line therapy. Plasma viral load was determined by an HIV-1 RNA assay and an HIV-RT assay in 629 samples (302 paired samples and 25 single time point samples at WK00) obtained from 327 patients. Overall, a strong correlation of r=0.96 was observed, with good correlation at WK00 (r=0.84) and at WK04 (r=0.77). Bland-Altman analysis of all samples showed a good level of agreement with a mean difference (bias) of 0.22 log10copies/mL. The performance of ExaVir Load V3 was not negatively affected by a nevirapine/efavirenz based antiretroviral regimen. The per test cost of measuring plasma viral load by the Abbott m2000rt and ExaVir Load V3 assays in a basic lab setting was $36.4 and $16.8, respectively. The strong correlation between the HIV-RT and HIV-1 RNA assays suggests that the HIV-RT assay can be an affordable alternative option for monitoring patients on antiretroviral therapy in resource-limited settings. ISRCTN79261738. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  1. Design and Testing of a Solid-Liquid Interface Monitor for High-Level Waste Tanks

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    McDaniel, D.; Awwad, A.; Roelant, D.

    2008-07-01

    A high-level waste (HLW) monitor has been designed, fabricated and tested at full-scale for deployment inside a Hanford tank. The Solid-Liquid Interface Monitor (SLIM) integrates a commercial sonar system with a mechanical deployment system for deploying into an underground waste tank. The system has undergone several design modifications based upon changing requirements at Hanford. We will present the various designs of the monitor from first to last and will present performance data from the various prototype systems. We will also present modeling of stresses in the enclosure under 85 mph wind loading. The system must be able to function atmore » winds up to 15 mph and must withstand a maximum loading of 85 mph. There will be several examples presented of engineering tradeoffs made as FIU analyzed new requirements and modified the design to accommodate. We will present our current plans for installing into the Cold Test Facility at Hanford and into a double-shelled tank at Hanford. Finally, we will present our vision for how this technology can be used at Hanford and Savannah River Site to improve the filling and emptying of high-level waste tanks. In conclusion: 1. The manually operated first-generation SLIM is a viable option on tanks where personnel are allowed to work on top of the tank. 2. The remote controlled second-generation SLIM can be utilized on tanks where personnel access is limited. 3. The totally enclosed fourth-generation SLIM, when the design is finalized, can be used when the possibility exists for wind dispersion of any HLW that maybe on the system. 4. The profiling sonar can be used effectively for real-time monitoring of the solid-liquid interface over a large area. (authors)« less

  2. Assessment of hazardous wastes for genotoxicity

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    DeMarini, D.M.; Houk, V.S.

    1987-09-01

    The authors have evaluated a group of short-term bioassays to identify those that may be suitable for screening large numbers of diverse hazardous industrial wastes for genotoxicity. Fifteen wastes (and dichloromethane extracts of these wastes) from a variety of manufacturing processes were tested for mutagenicity in Salmonella typhimurium strains TA98 and TA100 with and without Aroclor 1254-induced rat-liver S9. Ten of these wastes were fed by gavage to F-344 male rats, and the raw urines were assayed for mutagenicity in the presence of beta-glucuronidase in strain TA98 with S9. Six of these urines were extracted by C18/methanol elution, incubated withmore » beta-glucuronidase, and evaluated in strain TA98 with S9 and beta-glucuronidase. Fourteen of the wastes were examined for their ability to induce prophage lambda in Escherichia coli in a microsuspension assay. A second set of wastes, consisting of four industrial wastes, were evaluated in Salmonella and in a series of mammalian cell assays to measure mutagenicity, cytogenetic effects, and transformation.« less

  3. Clinical utility of the ARCHITECT HCV Ag assay for early treatment monitoring in patients with chronic hepatitis C genotype 1 infection.

    PubMed

    Vermehren, Johannes; Susser, Simone; Berger, Annemarie; Perner, Dany; Peiffer, Kai-Henrik; Allwinn, Regina; Zeuzem, Stefan; Sarrazin, Christoph

    2012-09-01

    Virologic response-monitoring is essential for determining therapy duration in patients with chronic hepatitis C virus (HCV) infection. This is usually performed using highly sensitive HCV-RNA assays. However, HCV-RNA assays are time-consuming, expensive and require highly trained personnel. Quantitative determination of HCV core-antigen (HCVAg) levels may be used to supplement treatment monitoring. The clinical utility of the ARCHITECT HCV Ag assay (Abbott Diagnostics) for response-guided therapy was investigated. We analyzed serum from 160 patients with HCV genotype 1 infection who had been treated with peg-interferon alfa-2b/ribavirin. HCVAg levels were determined at baseline, weeks 1, 2, 4 and 12. HCVAg levels were compared to those obtained with HCV-RNA assays: VERSANT HCV Quantitative 3.0 (bDNA) and Qualitative (TMA, both Siemens Healthcare) assay and the Abbott RealTime HCV assay (ART; Abbott Diagnostics). Baseline HCVAg levels correlated well with HCV-RNA as assessed by bDNA (r=0.91; p<0.0001) and ART (r=0.92; p<0.0001), respectively. Patients with undetectable HCVAg levels at week 1 had a 90.9% probability (positive predictive value) to achieve a rapid virologic response (HCV-RNA undetectable at week 4) based on TMA and 86.4% based on ART, respectively. Patients with less than 1 log(10) reduction in HCVAg between baseline and week 12 had a 90% probability (negative predictive value) to achieve a nonresponse (<2 log(10) decline in HCV-RNA between baseline and week 12) based on bDNA and 100% based on ART, respectively. Determination of HCVAg may be useful for antiviral response-monitoring in patients with HCV genotype 1 infection. Copyright © 2012 Elsevier B.V. All rights reserved.

  4. Detection and Monitoring of E-Waste Contamination through Remote Sensing and Image Analysis

    NASA Astrophysics Data System (ADS)

    Garb, Yaakov; Friedlander, Lonia

    2015-04-01

    Electronic waste (e-waste) is one of today's fastest growing waste streams, and also one of the more problematic, as this end-of-life product contains precious metals mixed with and embedded in a variety of low value and potentially harmful plastic and other materials. This combination creates a powerful incentive for informal value chains that transport, extract from, and dispose of e-waste materials in far-ranging and unregulated ways, and especially in settings where regulation and livelihood alternatives are sparse, most notably in areas of India, China, and Africa. E-waste processing is known to release a variety of contaminants, such as heavy metals and persistent organic pollutants, including flame retardants, dioxins and furans. In several sites, where the livelihoods of entire communities are dependent on e-waste processing, the resulting contaminants have been demonstrated to enter the hydrological system and food chain and have serious health and ecological effects. In this paper we demonstrate for the first time the usefulness of multi-spectral remote sensing imagery to detect and monitor the release and possibly the dispersal of heavy metal contaminants released in e-waste processing. While similar techniques have been used for prospecting or for studying heavy metal contamination from mining and large industrial facilities, we suggest that these techniques are of particular value in detecting contamination from the more dispersed, shifting, and ad-hoc kinds of release typical of e-waste processing. Given the increased resolution and decreased price of multi-spectral imagery, such techniques may offer a remarkably cost-effective and rapidly responsive means of assessing and monitoring this kind of contamination. We will describe the geochemical and multi-spectral image-processing principles underlying our approach, and show how we have applied these to an area in which we have a detailed, multi-temporal, spatially referenced, and ground

  5. Area-under-the-curve monitoring of cyclosporine therapy: Performance of different assay methods and their target concentrations

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Grevel, J.; Napoli, K.L.; Gibbons, S.

    1990-01-01

    The measurement of areas under the concentration-time curve (AUC) was recently introduced as an alternative to trough level monitoring of cyclosporine therapy. The AUC is divided by the oral dosing interval to calculate an average concentration. All measurements are performed at clinical steady state. The initial evaluation of AUC monitoring showed advantages over trough level monitoring with concentrations of cyclosporine measured in serum by the polyclonal radioimmunoassay of Sandoz. This assay technique is no longer available and the following assays were performed in parallel during up to 173 AUC determinations in 51 consecutive renal transplant patients: polyclonal fluorescence polarization immunoassaymore » of Abbott in serum, specific and nonspecific monoclonal radioimmunoassays using {sup 3}H and {sup 125}I tracers in serum and whole blood, and high performance liquid chromatography in whole blood. Both trough levels and average concentrations at steady state measured by those different techniques were significantly correlated with the oral dose. The best correlation (r2 = 0.54) was shown by average concentrations measured in whole blood by the specific monoclonal radioimmunoassay of Sandoz ({sup 3}H tracer). This monitoring technique was also associated with the smallest absolute error between repeated observations in the same patient while the oral dose rate remained the same or was changed. Both allegedly specific monoclonal radioimmunoassays (with {sup 3}H and {sup 125}I tracer) measured significantly higher concentrations than the liquid chromatography.« less

  6. Evaluation of New Quantitative Assays for Diagnosis and Monitoring of Cytomegalovirus Disease in Human Immunodeficiency Virus-Positive Patients

    PubMed Central

    Pellegrin, Isabelle; Garrigue, Isabelle; Binquet, Christine; Chene, Genevieve; Neau, Didier; Bonot, Pascal; Bonnet, Fabrice; Fleury, Herve; Pellegrin, Jean-Luc

    1999-01-01

    Cobas Amplicor CMV Monitor (CMM) and Quantiplex CMV bDNA 2.0 (CMV bDNA 2.0), two new standardized and quantitative assays for the detection of cytomegalovirus (CMV) DNA in plasma and peripheral blood leukocytes (PBLs), respectively, were compared to the CMV viremia assay, pp65 antigenemia assay, and the Amplicor CMV test (P-AMP). The CMV loads were measured in 384 samples from 58 human immunodeficiency virus (HIV) type 1-infected, CMV-seropositive subjects, including 13 with symptomatic CMV disease. The assays were highly concordant (agreement, 0.88 to 0.97) except when the CMV load was low. Quantitative results for plasma and PBLs were significantly correlated (Spearman ρ = 0.92). For PBLs, positive results were obtained 125 days before symptomatic CMV disease by CMV bDNA 2.0 and 124 days by pp65 antigenemia assay, whereas they were obtained 46 days before symptomatic CMV disease by CMM and P-AMP. At the time of CMV disease diagnosis, the sensitivity, specificity, and positive and negative predictive values of CMV bDNA 2.0 were 92.3, 97.8, 92.3, and 97.8%, respectively, whereas they were 92.3, 93.3, 80, and 97.8%, respectively, for the pp65 antigenemia assay; 84.6, 100, 100, and 95.7%, respectively, for CMM; and 76.9, 100, 100, and 93.8%, respectively, for P-AMP. Considering the entire follow-up, the sensitivity, specificity, and positive and negative predictive values of CMV bDNA 2.0 were 92.3, 73.3, 52.1, and 97.1%, respectively, whereas they were 100, 55.5, 39.4, and 100%, respectively, for the pp65 antigenemia assay; 92.3, 86.7, 66.7, and 97.5%, respectively, for CMM; and 84.6, 91.1, 73.3, and 95.3%, respectively, for P-AMP. Detection of CMV in plasma is technically easy and, despite its later positivity (i.e., later than in PBLs), can provide enough information sufficiently early so that HIV-infected patients can be effectively treated. In addition, these standardized quantitative assays accurately monitor the efficacy of anti-CMV treatment. PMID:10488165

  7. Evaluation of anaerobic co-digestion of dairy manure with food wastes via bio-methane potential assay and CSTR reactor.

    PubMed

    Ye, Yulin; Zamalloa, Carlos; Lin, Hongjian; Yan, Mi; Schmidt, David; Hu, Bo

    2015-01-01

    The introduction of food wastes into anaerobic digestion (AD) brings a promising scenario of increasing feedstock availability and overall energy production from AD. This study evaluated the biodegradability and methane potential from co-digestion of two typical food wastes, kitchen waste and chicken fat, with dairy manure. For single substrate, the bio-methane potential assays showed that kitchen waste had the highest methane yield of 352 L-CH4 kg(-1)-VS added, 92% more than dairy manure alone. Chicken fat at the same Volatile Solid (VS) level (2 g L(-1)) inhibited bio-methane production. Addition of kitchen waste and chicken fat to a VS percentage of up to 40% improved overall methane yield by 44% and 34%, respectively. Synergistic effect was observed when either combining two or three substrates as AD feedstock, possibly as a result of increased biodegradability of organic materials in chicken fat and kitchen waste compared with dairy manure. Addition of chicken fat improved methane yield more than kitchen waste. However, addition of chicken fat VS over 0.8 g L(-1) should be cautiously done because it may cause reactor failure due to decrease in pH. The maximum methane yield was 425 L-CH4 kg(-1)-VS, achieved at a VS ratio of 2:2:1 for kitchen waste, chicken fat, and dairy manure. Results from batch AD experiment demonstrated that supplementing dairy manure to chicken fat and/or kitchen waste improved alkalinity of substrate due to the inclusion of more titratable bases in dairy manure, and therefore stabilized the methanogenesis and substantially improved biogas yield. A mixture of substrates of kitchen waste, chicken fat, and dairy manure at a ratio of 1:1:3 was fed to a continuously stirred tank reactor operated at organic loading rates of 3.28, 6.55, and 2.18 g-COD L(-1)-day (hydraulic retention time of 20, 10, and 30 days, respectively) under mesophilic condition, and methane production rate reached 0.65, 0.95, and 0.34 L-CH4 L(-1)-reactor-day.

  8. Nevada National Security Site 2013 Waste Management Monitoring Report Area 3 and Area 5 Radioactive Waste Management Site

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hudson, D. B.

    Environmental monitoring data are collected at and around the Area 3 and Area 5 Radioactive Waste Management Sites (RWMSs) within the Nevada National Security Site (NNSS). These data are associated with radiation exposure, air, groundwater, meteorology, and vadose zone. This report summarizes the 2013 environmental data to provide an overall evaluation of RWMS performance and to support environmental compliance and performance assessment (PA) activities. Some of these data (e.g., radiation exposure, air, and groundwater) are presented in other reports (National Security Technologies, LLC, 2013; 2014a; 2014b). Direct radiation monitoring data indicate exposure levels at the RWMSs are within the rangemore » of background levels measured at the NNSS. Slightly elevated exposure levels outside the Area 3 RWMS are attributed to nearby historical aboveground nuclear weapons tests. Air monitoring data show tritium concentrations in water vapor and americium and plutonium concentrations in air particles are close to detection limits and background levels. The measured levels of radionuclides in air particulates and moisture are below Derived Concentration Standards for these radionuclides. Groundwater monitoring data indicate the groundwater in the uppermost aquifer beneath the Area 5 RWMS is not impacted by RWMS operations. Results of groundwater analysis from wells around the Area 5 RWMS were all below established investigation levels. Leachate samples collected from the leachate collection system at the mixed low-level waste cell were below established contaminant regulatory limits. The 105.8 millimeters (mm) (4.17 inches [in.]) of precipitation at the Area 3 RWMS during 2013 is 30% below the average of 150.3 mm (5.92 in.), and the 117.5 mm (4.63 in.) of precipitation at the Area 5 RWMS during 2013 is 5% below the average of 123.6 mm (4.86 in.). Water balance measurements indicate that evapotranspiration from the vegetated weighing lysimeter dries the soil and prevents

  9. Nevada National Security Site 2012 Waste Management Monitoring Report Area 3 and Area 5 Radioactive Waste Management Site

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hudson, David B.

    2013-09-10

    Environmental monitoring data are collected at and around the Area 3 and Area 5 Radioactive Waste Management Sites (RWMSs) at the Nevada National Security Site (NNSS). These data are associated with radiation exposure, air, groundwater, meteorology, and vadose zone. This report summarizes the 2012 environmental data to provide an overall evaluation of RWMS performance and to support environmental compliance and performance assessment (PA) activities. Some of these data (e.g., radiation exposure, air, and groundwater) are presented in other reports (National Security Technologies, LLC, 2012; 2013a; 2013b). Direct radiation monitoring data indicate exposure levels at the RWMSs are within the rangemore » of background levels measured at the NNSS. Slightly elevated exposure levels outside the Area 3 RWMS are attributed to nearby historical aboveground nuclear weapons tests. Air monitoring data show tritium concentrations in water vapor and americium and plutonium concentrations in air particles are only slightly above detection limits and background levels. The measured levels of radionuclides in air particulates and moisture are below Derived Concentration Standards for these radionuclides. Groundwater monitoring data indicate the groundwater in the uppermost aquifer beneath the Area 5 RWMS is not impacted by RWMS operations. Results of groundwater analysis from wells around the Area 5 RWMS were all below established investigation levels. Leachate samples collected from the leachate collection system at the mixed low-level waste cell were below established contaminant regulatory limits. The 133.9 millimeters (mm) (5.27 inches [in.]) of precipitation at the Area 3 RWMS during 2012 is 12% below the average of 153.0 mm (6.02 in.), and the 137.6 mm (5.42 in.) of precipitation at the Area 5 RWMS during 2012 is 11% below the average of 122.4 mm (4.82 in.). Water balance measurements indicate that evapotranspiration from the vegetated weighing lysimeter dries the soil and

  10. US Environmental rotection Agency's strategy for ground-water-quality monitoring at hazardous-waste land-disposal facilities located in karst terranes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Field, M.S.

    1988-11-01

    Ground-water monitoring of hazardous-waste land-disposal units by a network of wells is ineffective when located in karstic terranes. The U.S. Environmental Protection Agency (EPA) is currently proposing to modify its current ground-water-quality monitoring requirement of one upgradient well and three downgradient wells for disposal units located in karstic terranes. The convergent nature of subsurface flow to cave streams in karstic terranes requires that effective monitoring wells intercept the cave streams. Wells located around a hazardous-waste disposal unit, but not in the specific cave stream draining the site, are only providing irrelevant data and a false sense of security because themore » water samples from such wells are not necessarily from the hazardous-waste disposal unit. A case study is provided in this paper. EPA is drafting a guidance document that will allow monitoring by wells, only if the up- and down-gradient wells can be demonstrated to be hydraulically connected by means of dye-trace studies. If not, then the monitoring of springs shown to be hydraulically connected to the facility by dye-tracing studies would be required. Monitoring for sinkhole development will also be required to provide advance warning of sinkhole collapse. The investigation and determination of the probability of sinkhole collapse is given special treatment.« less

  11. Opportunities and challenges for structural health monitoring of radioactive waste systems and structures

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Giurgiutiu, Victor; Mendez Torres, Adrian E.

    2013-07-01

    Radioactive waste systems and structures (RWSS) are safety-critical facilities in need of monitoring over prolonged periods of time. Structural health monitoring (SHM) is an emerging technology that aims at monitoring the state of a structure through the use of networks of permanently mounted sensors. SHM technologies have been developed primarily within the aerospace and civil engineering communities. This paper addresses the issue of transitioning the SHM concept to the monitoring of RWSS and evaluates the opportunities and challenges associated with this process. Guided wave SHM technologies utilizing structurally-mounted piezoelectric wafer active sensors (PWAS) have a wide range of applications basedmore » on both propagating-wave and standing-wave methodologies. Hence, opportunities exist for transitioning these SHM technologies into RWSS monitoring. However, there exist certain special operational conditions specific to RWSS such as: radiation field, caustic environments, marine environments, and chemical, mechanical and thermal stressors. In order to address the high discharge of used nuclear fuel (UNF) and the limited space in the storage pools the U.S. the Department of Energy (DOE) has adopted a 'Strategy for the Management and Disposal of Used Nuclear Fuel and High-Level Radioactive Waste' (January 2013). This strategy endorses the key principles that underpin the Blue Ribbon Commission's on America's Nuclear Future recommendations to develop a sustainable program for deploying an integrated system capable of transporting, storing, and disposing of UNF and high-level radioactive waste from civilian nuclear power generation, defense, national security, and other activities. This will require research to develop monitoring, diagnosis, and prognosis tools that can aid to establish a strong technical basis for extended storage and transportation of UNF. Monitoring of such structures is critical for assuring the safety and security of the nation's spent

  12. The Effects of Data Collection Method and Monitoring of Workers’ Behavior on the Generation of Demolition Waste

    PubMed Central

    Cha, Gi-Wook; Kim, Young-Chan; Moon, Hyeun Jun; Hong, Won-Hwa

    2017-01-01

    The roles of both the data collection method (including proper classification) and the behavior of workers on the generation of demolition waste (DW) are important. By analyzing the effect of the data collection method used to estimate DW, and by investigating how workers’ behavior can affect the total amount of DW generated during an actual demolition process, it was possible to identify strategies that could improve the prediction of DW. Therefore, this study surveyed demolition waste generation rates (DWGRs) for different types of building by conducting on-site surveys immediately before demolition in order to collect adequate and reliable data. In addition, the effects of DW management strategies and of monitoring the behavior of workers on the actual generation of DW were analyzed. The results showed that when monitoring was implemented, the estimates of DW obtained from the DWGRs that were surveyed immediately before demolition and the actual quantities of DW reported by the demolition contractors had an error rate of 0.63% when the results were compared. Therefore, this study has shown that the proper data collection method (i.e., data were collected immediately before demolition) applied in this paper and monitoring on the demolition site have a significant impact on waste generation. PMID:29023378

  13. What level of estrogenic activity determined by in vitro assays in municipal waste waters can be considered as safe?

    PubMed

    Jarošová, Barbora; Bláha, Luděk; Giesy, John P; Hilscherová, Klára

    2014-03-01

    In vitro assays are broadly used tools to evaluate the estrogenic activity in Waste Water Treatment Plant (WWTP) effluents and their receiving rivers. Since potencies of individual estrogens to induce in vitro and in vivo responses can differ it is not possible to directly evaluate risks based on in vitro measures of estrogenic activity. Estrone, 17beta-estradiol, 17alfa-ethinylestradiol and to some extent, estriol have been shown to be responsible for the majority of in vitro estrogenic activity of municipal WWTP effluents. Therefore, in the present study safe concentrations of Estrogenic Equivalents (EEQs-SSE) in municipal WWTP effluents were derived based on simplified assumption that the steroid estrogens are responsible for all estrogenicity determined with particular in vitro assays. EEQs-SSEs were derived using the bioassay and testing protocol-specific in vitro potencies of steroid estrogens, in vivo predicted no effect concentration (PNECs) of these compounds, and their relative contributions to the overall estrogenicity detected in municipal WWTP effluents. EEQs-SSEs for 15 individual bioassays varied from 0.1 to 0.4ng EEQ/L. The EEQs-SSEs are supposed to be increased by use of location-specific dilution factors of WWTP effluents entering receiving rivers. They are applicable to municipal wastewater and rivers close to their discharges, but not to industrial waste waters. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Real-time monitoring of enzyme-free strand displacement cascades by colorimetric assays.

    PubMed

    Duan, Ruixue; Wang, Boya; Hong, Fan; Zhang, Tianchi; Jia, Yongmei; Huang, Jiayu; Hakeem, Abdul; Liu, Nannan; Lou, Xiaoding; Xia, Fan

    2015-03-19

    The enzyme-free toehold-mediated strand displacement reaction has shown potential for building programmable DNA circuits, biosensors, molecular machines and chemical reaction networks. Here we report a simple colorimetric method using gold nanoparticles as signal generators for the real-time detection of the product of the strand displacement cascade. During the process the assembled gold nanoparticles can be separated, resulting in a color change of the solution. This assay can also be applied in complex mixtures, fetal bovine serum, and to detect single-base mismatches. These results suggest that this method could be of general utility to monitor more complex enzyme-free strand displacement reaction-based programmable systems or for further low-cost diagnostic applications.

  15. Quantitative Validation of the Presto Blue Metabolic Assay for Online Monitoring of Cell Proliferation in a 3D Perfusion Bioreactor System.

    PubMed

    Sonnaert, Maarten; Papantoniou, Ioannis; Luyten, Frank P; Schrooten, Jan Ir

    2015-06-01

    As the fields of tissue engineering and regenerative medicine mature toward clinical applications, the need for online monitoring both for quantitative and qualitative use becomes essential. Resazurin-based metabolic assays are frequently applied for determining cytotoxicity and have shown great potential for monitoring 3D bioreactor-facilitated cell culture. However, no quantitative correlation between the metabolic conversion rate of resazurin and cell number has been defined yet. In this work, we determined conversion rates of Presto Blue, a resazurin-based metabolic assay, for human periosteal cells during 2D and 3D static and 3D perfusion cultures. Our results showed that for the evaluated culture systems there is a quantitative correlation between the Presto Blue conversion rate and the cell number during the expansion phase with no influence of the perfusion-related parameters, that is, flow rate and shear stress. The correlation between the cell number and Presto Blue conversion subsequently enabled the definition of operating windows for optimal signal readouts. In conclusion, our data showed that the conversion of the resazurin-based Presto Blue metabolic assay can be used as a quantitative readout for online monitoring of cell proliferation in a 3D perfusion bioreactor system, although a system-specific validation is required.

  16. Quantitative Validation of the Presto Blue™ Metabolic Assay for Online Monitoring of Cell Proliferation in a 3D Perfusion Bioreactor System

    PubMed Central

    Sonnaert, Maarten; Papantoniou, Ioannis; Luyten, Frank P.

    2015-01-01

    As the fields of tissue engineering and regenerative medicine mature toward clinical applications, the need for online monitoring both for quantitative and qualitative use becomes essential. Resazurin-based metabolic assays are frequently applied for determining cytotoxicity and have shown great potential for monitoring 3D bioreactor-facilitated cell culture. However, no quantitative correlation between the metabolic conversion rate of resazurin and cell number has been defined yet. In this work, we determined conversion rates of Presto Blue™, a resazurin-based metabolic assay, for human periosteal cells during 2D and 3D static and 3D perfusion cultures. Our results showed that for the evaluated culture systems there is a quantitative correlation between the Presto Blue conversion rate and the cell number during the expansion phase with no influence of the perfusion-related parameters, that is, flow rate and shear stress. The correlation between the cell number and Presto Blue conversion subsequently enabled the definition of operating windows for optimal signal readouts. In conclusion, our data showed that the conversion of the resazurin-based Presto Blue metabolic assay can be used as a quantitative readout for online monitoring of cell proliferation in a 3D perfusion bioreactor system, although a system-specific validation is required. PMID:25336207

  17. 40 CFR 62.15380 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... curtain incinerators that burn 100 percent yard waste? 62.15380 Section 62.15380 Protection of Environment... Combustion Units Constructed on or Before August 30, 1999 Air Curtain Incinerators That Burn 100 Percent Yard Waste § 62.15380 How must I monitor opacity for air curtain incinerators that burn 100 percent yard...

  18. 40 CFR 62.15380 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... curtain incinerators that burn 100 percent yard waste? 62.15380 Section 62.15380 Protection of Environment... Combustion Units Constructed on or Before August 30, 1999 Air Curtain Incinerators That Burn 100 Percent Yard Waste § 62.15380 How must I monitor opacity for air curtain incinerators that burn 100 percent yard...

  19. 40 CFR 62.15380 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... curtain incinerators that burn 100 percent yard waste? 62.15380 Section 62.15380 Protection of Environment... Combustion Units Constructed on or Before August 30, 1999 Air Curtain Incinerators That Burn 100 Percent Yard Waste § 62.15380 How must I monitor opacity for air curtain incinerators that burn 100 percent yard...

  20. 40 CFR 62.15380 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... curtain incinerators that burn 100 percent yard waste? 62.15380 Section 62.15380 Protection of Environment... Combustion Units Constructed on or Before August 30, 1999 Air Curtain Incinerators That Burn 100 Percent Yard Waste § 62.15380 How must I monitor opacity for air curtain incinerators that burn 100 percent yard...

  1. 40 CFR 62.15380 - How must I monitor opacity for air curtain incinerators that burn 100 percent yard waste?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... curtain incinerators that burn 100 percent yard waste? 62.15380 Section 62.15380 Protection of Environment... Combustion Units Constructed on or Before August 30, 1999 Air Curtain Incinerators That Burn 100 Percent Yard Waste § 62.15380 How must I monitor opacity for air curtain incinerators that burn 100 percent yard...

  2. U.S. EPA'S STRATEGY FOR GROUND WATER QUALITY MONITORING AT HAZARDOUS WASTE LAND DISPOSAL FACILITIES LOCATED IN KARST TERRANES

    EPA Science Inventory

    Ground water monitoring of hazardous waste land disposal units by a network of wells is ineffective when located in karstic terranes. The U.S. Environmental Protection Agency (EPA) is currently proposing to modify its current ground water quality monitoring requirement of one upg...

  3. Comparison of the Quantiplex Version 3.0 Assay and a Sensitized Amplicor Monitor Assay for Measurement of Human Immunodeficiency Virus Type 1 RNA Levels in Plasma Samples

    PubMed Central

    Highbarger, Helene C.; Alvord, W. Gregory; Jiang, Min Kang; Shah, Akram S.; Metcalf, Julia A.; Lane, H. Clifford; Dewar, Robin L.

    1999-01-01

    This study evaluated correlation and agreement between version 3 of the Quantiplex human immunodeficiency virus type 1 (HIV-1) RNA assay (v3 branched DNA [bDNA]) and a sensitized Amplicor HIV-1 Monitor assay (reverse transcription [RT]-PCR) for the measurement of HIV RNA. Three hundred eighteen samples from 59 randomly selected, HIV-1-seropositive persons on various drug protocols from the National Institute of Allergy and Infectious Diseases HIV outpatient clinic were studied. The results indicate that v3 bDNA and RT-PCR are highly correlated (r = 0.98) and are in good agreement (mean difference in log10 copies/ml ± 2 standard deviations = 0.072 ± 0.371). The relationship between values obtained by both assays is given by the following equation: log10v3 bDNA = −0.0915 + 1.0052 · log10RT-PCR. This represents a 1.026-fold difference between log10RT-PCR values and log10v3 bDNA values. PMID:10523562

  4. The Mutation-Associated Neoantigen Functional Expansion of Specific T cells (MANAFEST) assay: a sensitive platform for monitoring antitumor immunity.

    PubMed

    Danilova, Ludmila; Anagnostou, Valsamo; Caushi, Justina X; Sidhom, John-William; Guo, Haidan; Chan, Hok Yee; Suri, Prerna; Tam, Ada J; Zhang, Jiajia; El Asmar, Margueritta; Marrone, Kristen A; Naidoo, Jarushka; Brahmer, Julie R; Forde, Patrick M; Baras, Alexander S; Cope, Leslie; Velculescu, Victor E; Pardoll, Drew; Housseau, Franck; Smith, Kellie N

    2018-06-12

    Mutation-associated neoantigens (MANAs) are a target of antitumor T-cell immunity. Sensitive, simple, and standardized assays are needed to assess the repertoire of functional MANA-specific T cells in oncology. Assays analyzing in vitro cytokine production such as ELISpot and intracellular cytokine staining (ICS) have been useful but have limited sensitivity in assessing tumor-specific T-cell responses and do not analyze antigen-specific T-cell repertoires. The FEST (Functional Expansion of Specific T cells) assay described herein integrates TCR sequencing of short-term, peptide-stimulated cultures with a bioinformatic platform to identify antigen-specific clonotypic amplifications. This assay can be adapted for all types of antigens, including mutation associated neoantigens (MANAs) via tumor exome-guided prediction of MANAs. Following in vitro identification by the MANAFEST assay, the MANA-specific CDR3 sequence can be used as a molecular barcode to detect and monitor the dynamics of these clonotypes in blood, tumor, and normal tissue of patients receiving immunotherapy. MANAFEST is compatible with high-throughput routine clinical and lab practices. Copyright ©2018, American Association for Cancer Research.

  5. Deformation Monitoring of Waste-Rock-Backfilled Mining Gob for Ground Control

    PubMed Central

    Zhao, Tongbin; Zhang, Yubao; Zhang, Zhenyu; Li, Zhanhai; Ma, Shuqi

    2017-01-01

    Backfill mining is an effective option to mitigate ground subsidence, especially for mining under surface infrastructure, such as buildings, dams, rivers and railways. To evaluate its performance, continual long-term field monitoring of the deformation of backfilled gob is important to satisfy strict public scrutiny. Based on industrial Ethernet, a real-time monitoring system was established to monitor the deformation of waste-rock-backfilled gob at −700 m depth in the Tangshan coal mine, Hebei Province, China. The designed deformation sensors, based on a resistance transducer mechanism, were placed vertically between the roof and floor. Stress sensors were installed above square steel plates that were anchored to the floor strata. Meanwhile, data cables were protected by steel tubes in case of damage. The developed system continually harvested field data for three months. The results show that industrial Ethernet technology can be reliably used for long-term data transmission in complicated underground mining conditions. The monitoring reveals that the roof subsidence of the backfilled gob area can be categorized into four phases. The bearing load of the backfill developed gradually and simultaneously with the deformation of the roof strata, and started to be almost invariable when the mining face passed 97 m. PMID:28475168

  6. Deformation Monitoring of Waste-Rock-Backfilled Mining Gob for Ground Control.

    PubMed

    Zhao, Tongbin; Zhang, Yubao; Zhang, Zhenyu; Li, Zhanhai; Ma, Shuqi

    2017-05-05

    Backfill mining is an effective option to mitigate ground subsidence, especially for mining under surface infrastructure, such as buildings, dams, rivers and railways. To evaluate its performance, continual long-term field monitoring of the deformation of backfilled gob is important to satisfy strict public scrutiny. Based on industrial Ethernet, a real-time monitoring system was established to monitor the deformation of waste-rock-backfilled gob at -700 m depth in the Tangshan coal mine, Hebei Province, China. The designed deformation sensors, based on a resistance transducer mechanism, were placed vertically between the roof and floor. Stress sensors were installed above square steel plates that were anchored to the floor strata. Meanwhile, data cables were protected by steel tubes in case of damage. The developed system continually harvested field data for three months. The results show that industrial Ethernet technology can be reliably used for long-term data transmission in complicated underground mining conditions. The monitoring reveals that the roof subsidence of the backfilled gob area can be categorized into four phases. The bearing load of the backfill developed gradually and simultaneously with the deformation of the roof strata, and started to be almost invariable when the mining face passed 97 m.

  7. Production of an innovative fertilizer from organic waste: process monitoring by hyperspectral imaging

    NASA Astrophysics Data System (ADS)

    Serranti, Silvia; Bonifazi, Giuseppe; Fabbri, Andrea; Dall'Ara, Alice; Garcia Izquierdo, Carlos

    2015-05-01

    The European directive 2008/98/CE establishes a legal framework for the treatment of waste within the Community. It aims at protecting the environment and human health through the prevention of the harmful effects of waste generation and waste management. In order to better protect the environment, the Member States should adopt measures for the treatment of their waste according to a hierarchy as outlined: prevention, preparing for reuse, recycling, energy recovery, disposal. In this context, the European project LIFE12 ENV/IT/000356 "RESAFE" is addressed to produce and utilize a new class of fertilizers characterized by reduced salinity in order to substitute chemical and mineral fertilizers through a technological route based on Urban Organic Waste (UOW), Farm Organic Residues (FOR), Bio-Chars (BC) and Vegetable Active Principles (VAP) processing. Following this approach, it will be possible for farmers and urban waste managers to reduce costs and to obtain environmental and economic incomes. Furthermore, environmental impacts will be also reduced contributing to decrease the greenhouse emissions from landfills and from the production of mineral fertilizers. In this paper, specific innovative sensing architectures, based on Hyper-Spectral Imaging (HSI) devices working in the near infrared (NIR) range, and related detection architectures, is presented and discussed in order to define and apply smart detection engines to follow the transformations of the complex material, resulting from UOW, FOR, BC and VAP based recipes during the different stages of the fertilizer production process. Results show as the fertilizer production process can be monitored adopting the NIR-HSI approach.

  8. Monitoring for a specific management objective: protection of shorebird foraging habitat adjacent to a waste water treatment plant.

    PubMed

    Morris, Liz; Petch, David; May, David; Steele, William K

    2017-05-01

    Intertidal invertebrates are often used in environmental monitoring programs as they are good indicators of water quality and an important food source for many species of fish and birds. We present data from a monitoring program where the primary aim is to report on the condition of the potential invertebrate prey abundance, biomass and diversity for migrating shorebirds on mudflats adjacent to a waste water treatment plant in a Ramsar listed wetland in Victoria, Australia. A key threat to the foraging habitat at this site has been assessed as a reduction in potential prey items as a result of the changes to the waste water treatment processes. We use control charts, which summarise data from intertidal mudflats across the whole shoreline of the adjacent waste water treatment plant, to elicit a management response when trigger levels are reached. We then examine data from replicate discharge and control sites to determine the most appropriate management response. The monitoring program sits within an adaptive management framework where management decisions are reviewed and the data is examined at different scales to evaluate and modify our models of the likely outcomes of management actions. This study provides a demonstration of the process undertaken in a year when trigger levels were reached and a management decision was required. This highlights the importance of monitoring data from a range of scales in reducing uncertainty and improving decision making in complex systems.

  9. Improvement of non-destructive fissile mass assays in α low-level waste drums: A matrix correction method based on neutron capture gamma-rays and a neutron generator

    NASA Astrophysics Data System (ADS)

    Jallu, F.; Loche, F.

    2008-08-01

    Within the framework of radioactive waste control, non-destructive assay (NDA) methods may be employed. The active neutron interrogation (ANI) method is now well-known and effective in quantifying low α-activity fissile masses (mainly 235U, 239Pu, 241Pu) with low densities, i.e. less than about 0.4, in radioactive waste drums of volumes up to 200 l. The PROMpt Epithermal and THErmal interrogation Experiment (PROMETHEE [F. Jallu, A. Mariani, C. Passard, A.-C. Raoux, H. Toubon, Alpha low level waste control: improvement of the PROMETHEE 6 assay system performances. Nucl. Technol. 153 (January) (2006); C. Passard, A. Mariani, F. Jallu, J. Romeyer-Dherber, H. Recroix, M. Rodriguez, J. Loridon, C. Denis, PROMETHEE: an alpha low level waste assay system using passive and active neutron measurement methods. Nucl. Technol. 140 (December) (2002) 303-314]) based on ANI has been under development since 1996 to reach the incinerating α low level waste (LLW) criterion of about 50 Bq[α] per gram of crude waste (≈50 μg Pu) in 118 l drums on the date the drums are conditioned. Difficulties arise when dealing with matrices containing neutron energy moderators such as H and neutron absorbents such as Cl. These components may have a great influence on the fissile mass deduced from the neutron signal measured by ANI. For example, the calibration coefficient measured in a 118 l drum containing a cellulose matrix (density d = 0.144 g cm -3) may be 50 times higher than that obtained in a poly-vinyl-chloride matrix ( d = 0.253 g cm -3). Without any information on the matrix, the fissile mass is often overestimated due to safety procedures and by considering the most disadvantageous calibration coefficient corresponding to the most absorbing and moderating calibration matrix. The work discussed in this paper was performed at the CEA Nuclear Measurement Laboratory in France. It concerns the development of a matrix effect correction method, which consists in identifying and quantifying

  10. Comparison of the Abbott m2000 HIV-1 Real-Time and Roche AMPLICOR Monitor v1.5 HIV-1 assays on plasma specimens from Rakai, Uganda.

    PubMed

    Ssebugenyi, I; Kizza, A; Mpoza, B; Aluma, G; Boaz, I; Newell, K; Laeyendecker, O; Shott, J P; Serwadda, D; Reynolds, S J

    2011-07-01

    The need for viral load (VL) monitoring of HIV patients receiving antiretroviral therapy (ART) in resource-limited settings (RLS) has become apparent with studies showing the limitations of immunological monitoring. We compared the Abbott m2000 Real-Time (Abbott) HIV-1 assay with the Roche AMPLICOR Monitor v1.5 (Roche) HIV-1 assay over a range of VL concentrations. Three hundred and eleven plasma samples were tested, including 164 samples from patients on ART ≥ six months and 147 from ART-naïve patients. The Roche assay detected ≥400 copies/mL in 158 (50.8%) samples. Of these, Abbott produced 145 (91.8%) detectable results ≥400 copies/mL; 13 (8.2%) samples produced discrepant results. Concordance between the assays for detecting HIV-1 RNA ≥400 copies/mL was 95.8% (298/311). The sensitivity, specificity, positive predictive value and negative predictive value of Abbott to detect HIV-1 RNA ≥400 copies/mL were 91.8%, 100%, 100% and 92.2%, respectively. For the 151 samples with HIV-1 RNA ≥400 copies/mL for both assays, a good linear correlation was found (r = 0.81, P < 0.0001; mean difference, 0.05). The limits of agreement were -0.97 and 1.07 log(10) copies/mL (mean ± 2 SD). The Abbott assay performed well in our setting, offering an alternative methodology for HIV-1 VL for laboratories with realtime polymerase chain reaction (PCR) capacity.

  11. MTS dye based colorimetric CTLL-2 cell proliferation assay for product release and stability monitoring of interleukin-15: assay qualification, standardization and statistical analysis.

    PubMed

    Soman, Gopalan; Yang, Xiaoyi; Jiang, Hengguang; Giardina, Steve; Vyas, Vinay; Mitra, George; Yovandich, Jason; Creekmore, Stephen P; Waldmann, Thomas A; Quiñones, Octavio; Alvord, W Gregory

    2009-08-31

    criteria based on comparability and consistency in the four parameters of the model. The assay is precise, accurate and robust and can be fully validated. Applications of the assay were established including process development support, release of the rHuIL-15 product for pre-clinical and clinical studies, and for monitoring storage stability.

  12. Effective solutions for monitoring the electrostatic separation of metal and plastic granular waste from electric and electronic equipment.

    PubMed

    Senouci, Khouira; Medles, Karim; Dascalescu, Lucian

    2013-02-01

    The variability of the quantity and purity of the recovered materials is a serious drawback for the application of electrostatic separation technologies to the recycling of granular wastes. In a series of previous articles we have pointed out how capability and classic control chart concepts could be employed for better mastering the outcome of such processes. In the present work, the multiple exponentially weighted moving average (MEWMA) control chart is introduced and shown to be more effective than the Hotelling T2 chart for monitoring slow varying changes in the electrostatic separation of granular mixtures originating from electric and electronic equipment waste. The operation of the industrial process was simulated by using a laboratory roll-type electrostatic separator and granular samples resulting from shredded electric cable wastes. The 25 tests carried out during the observation phase enabled the calculation of the upper and lower control limits for the two control charts considered in the present study. The 11 additional tests that simulated the monitoring phase pointed out that the MEWMA chart is more effective than Hotelling's T(2) chart in detecting slow varying changes in the outcome of a process. As the reverse is true in the case of abrupt alterations of monitored process performances, simultaneous usage of the two control charts is strongly recommended. While this study focused on a specific electrostatic separation process, using the MEWMA chart together with the well known Hotelling's T(2) chart should be applicable to the statistical control of other complex processes in the field of waste processing.

  13. Acute toxicity assessment of Polish (waste) water with a microplate-based Hydra attenuata assay: a comparison with the Microtox test.

    PubMed

    Pardos, M; Benninghoff, C; Guéguen, C; Thomas, R; Dobrowolski, J; Dominik, J

    1999-12-15

    The use of Hydra attenuata in acute toxicity assessment is a potentially useful tool in (waste) water biomonitoring. The purpose of this study was to compare the sensitivity of H. attenuata with the extensively used Microtox test on 14 (waste) water samples from the Kraków region (South Poland). To this end, specific morphological changes displayed by the freshwater cnidarian Hydra attenuata (lethal LC50s and sublethal EC50s effects) and bioluminescence of the marine bacteria Vibrio fisheri (Microtox) were compared. Clearly, the Hydra assay was the more sensitive indicator of toxicity. No relationship was found among Hydra toxicological responses and water levels of As, Cd, Co, Cu, Pb and Zn. However, it appeared that toxicity to Hydra might be due to ammonia levels. Additional studies to better circumscribe the tolerance of H. attenuata to 'natural' water characteristics are needed.

  14. Metabolic response of environmentally isolated microorganisms to industrial effluents: Use of a newly described cell culture assay

    NASA Technical Reports Server (NTRS)

    Ferebee, Robert N.

    1992-01-01

    An environmental application using a microtiter culture assay to measure the metabolic sensitivity of microorganisms to petrochemical effluents will be tested. The Biomedical Operations and Research Branch at NASA JSC has recently developed a rapid and nondestructive method to measure cell growth and metabolism. Using a colorimetric procedure the uniquely modified assay allows the metabolic kinetics of prokaryotic and eukaryotic cells to be measured. Use of such an assay if adapted for the routine monitoring of waste products, process effluents, and environmentally hazardous substances may prove to be invaluable to the industrial community. The microtiter method as described will be tested using microorganisms isolated from the Galveston Bay aquatic habitat. The microbial isolates will be identified prior to testing using the automated systems available at JSC. Sodium dodecyl sulfate (SDS), cadmium, and lead will provide control toxic chemicals. The toxicity of industrial effluent from two industrial sites will be tested. An effort will be made to test the efficacy of this assay for measuring toxicity in a mixed culture community.

  15. Real Time Monitoring System of Pollution Waste on Musi River Using Support Vector Machine (SVM) Method

    NASA Astrophysics Data System (ADS)

    Fachrurrozi, Muhammad; Saparudin; Erwin

    2017-04-01

    Real-time Monitoring and early detection system which measures the quality standard of waste in Musi River, Palembang, Indonesia is a system for determining air and water pollution level. This system was designed in order to create an integrated monitoring system and provide real time information that can be read. It is designed to measure acidity and water turbidity polluted by industrial waste, as well as to show and provide conditional data integrated in one system. This system consists of inputting and processing the data, and giving output based on processed data. Turbidity, substances, and pH sensor is used as a detector that produce analog electrical direct current voltage (DC). Early detection system works by determining the value of the ammonia threshold, acidity, and turbidity level of water in Musi River. The results is then presented based on the level group pollution by the Support Vector Machine classification method.

  16. Comparison of quantitative cytomegalovirus (CMV) PCR in plasma and CMV antigenemia assay: clinical utility of the prototype AMPLICOR CMV MONITOR test in transplant recipients.

    PubMed

    Caliendo, A M; St George, K; Kao, S Y; Allega, J; Tan, B H; LaFontaine, R; Bui, L; Rinaldo, C R

    2000-06-01

    The correlation between the prototype AMPLICOR CMV MONITOR test (Roche Molecular Systems), a quantitative PCR assay, and the cytomegalovirus (CMV) pp65 antigenemia assay was evaluated in transplant recipients. Sequential blood specimens were collected on 29 patients (491 specimens), the leukocyte fraction was tested by CMV antigenemia, and quantitative PCR was performed on plasma specimens. None of the 15 patients (242 specimens) who were antigenemia negative were positive for CMV DNA by PCR, and none of these patients developed active CMV disease. There were 14 antigenemia-positive patients, 8 of whom developed active CMV disease. In all patients, there was a good association between the antigenemia and PCR assays. Ganciclovir-resistant virus was isolated from three patients with active CMV disease. These three patients had persistently elevated levels of antigenemia and CMV DNA by PCR when resistance to ganciclovir developed. This standardized, quantitative CMV PCR assay on plasma has clinical utility for the diagnosis of active disease and in monitoring the response to antiviral therapy in transplant recipients.

  17. Comparison of Quantitative Cytomegalovirus (CMV) PCR in Plasma and CMV Antigenemia Assay: Clinical Utility of the Prototype AMPLICOR CMV MONITOR Test in Transplant Recipients

    PubMed Central

    Caliendo, Angela M.; St. George, Kirsten; Kao, Shaw-Yi; Allega, Jessica; Tan, Ban-Hock; LaFontaine, Robert; Bui, Larry; Rinaldo, Charles R.

    2000-01-01

    The correlation between the prototype AMPLICOR CMV MONITOR test (Roche Molecular Systems), a quantitative PCR assay, and the cytomegalovirus (CMV) pp65 antigenemia assay was evaluated in transplant recipients. Sequential blood specimens were collected on 29 patients (491 specimens), the leukocyte fraction was tested by CMV antigenemia, and quantitative PCR was performed on plasma specimens. None of the 15 patients (242 specimens) who were antigenemia negative were positive for CMV DNA by PCR, and none of these patients developed active CMV disease. There were 14 antigenemia-positive patients, 8 of whom developed active CMV disease. In all patients, there was a good association between the antigenemia and PCR assays. Ganciclovir-resistant virus was isolated from three patients with active CMV disease. These three patients had persistently elevated levels of antigenemia and CMV DNA by PCR when resistance to ganciclovir developed. This standardized, quantitative CMV PCR assay on plasma has clinical utility for the diagnosis of active disease and in monitoring the response to antiviral therapy in transplant recipients. PMID:10834964

  18. Sense and sensibility: the use of cell death biomarker assays in high-throughput anticancer drug screening and monitoring treatment responses.

    PubMed

    Shoshan, Maria C; Havelka, Associate Professor Principal Investigator Aleksandra Mandic; Neumann, Frank; Linder, Stig

    2006-11-01

    Cell-based screening allows identification of biologically active compounds, for example, potential anticancer drugs. In this review, various screening assays are discussed in terms of what they measure and how this affects interpretation and relevance. High-throughput (HT) assays of viability based on the reduction of exogenous substrates do not always reflect viability or cell number levels. Membrane integrity assays can be used for HT quantification of cell death, but are non-specific as to the death mode. Several HT assays monitor end point apoptosis. Screening libraries at a single concentration (micromolar) can prevent detection of potent apoptosis inducers, as high concentrations may induce mainly necrosis. Using monolayer cultures limits the significance of cell-based screening as the properties of monolayer cells differ from tumours in vivo. Spheroid cultures are more physiological, but are impractical for screening by conventional methods. The authors have developed an assay quantifying accumulation of a caspase-cleaved protein specific for epithelial cells. It provides an integrated measure of apoptosis in two- and three-dimensional cultures and can be used as a blood biomarker assay for tumour apoptosis in vivo.

  19. Nevada National Security Site 2015 Waste Management Monitoring Report Area 3 and Area 5 Radioactive Waste Management Sites

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Black, David; Hudson, David

    Environmental monitoring data are collected at and around the Area 3 and Area 5 Radioactive Waste Management Sites (RWMSs) within the Nevada National Security Site (NNSS). These data include direct radiation exposure, as well as radiation from the air, groundwater, meteorology, and vadose zone. This report summarizes the 2015 environmental data to provide an overall evaluation of RWMS performance and to support environmental compliance and performance assessment (PA) activities. Some of these data (e.g., radiation exposure, air, and groundwater) are presented in other reports developed by National Security Technologies, LLC. Direct radiation monitoring data indicate exposure levels at the RWMSsmore » are within the range of background levels measured at the NNSS. Slightly elevated exposure levels outside the Area 3 RWMS are attributed to nearby historical aboveground nuclear weapons tests. Air monitoring data show that tritium concentrations in water vapor and americium and plutonium concentrations in air particles are below Derived Concentration Standards for these radionuclides. Groundwater monitoring data indicate the groundwater in the uppermost aquifer beneath the Area 5 RWMS is not impacted by RWMS operations. Results of groundwater analysis from wells around the Area 5 RWMS were all below established investigation levels. Leachate samples collected from the leachate collection system at the mixed low-level waste cell were below established contaminant regulatory limits. During 2015, precipitation at the Area 3 RWMS was 0.9% above average, and precipitation at the Area 5 RWMS was 25% above average. Water balance measurements indicate that evapotranspiration from the vegetated weighing lysimeter dries the soil and prevents downward percolation of precipitation more effectively than evaporation as measured from the bare-soil weighing lysimeter. The 1.8 inches of precipitation in September reached the lowest sensors at 180 cm on the Cell 5S and 5N covers

  20. Evaluation of the COBAS Hepatitis C Virus (HCV) TaqMan analyte-specific reagent assay and comparison to the COBAS Amplicor HCV Monitor V2.0 and Versant HCV bDNA 3.0 assays.

    PubMed

    Konnick, Eric Q; Williams, Sheri M; Ashwood, Edward R; Hillyard, David R

    2005-05-01

    Performance characteristics of the COBAS hepatitis C virus (HCV) TaqMan analyte-specific reagent (TM-ASR) assay using the QIAGEN BioRobot 9604 for RNA extraction were evaluated and compared to the COBAS Amplicor HCV Monitor V2.0 (Amplicor) and Versant HCV bDNA 3.0 (Versant) assays using clinical samples. Calibration of TM-ASR using Armored RNA allowed determination of the distribution of HCV RNA in clinical samples, using 22,399 clinical samples. Limit of detection, linearity, and inter- and intraassay assay precision were determined for the TM-ASR assay using multiple clinical specimen panels across multiple determinations. Genotype specificity for the TM-ASR assay was determined using samples with different HCV RNA genotypes evaluated and compared against predetermined results. Contamination control of the TM-ASR assay was evaluated using pools of HCV RNA-positive and -negative samples tested in a checkerboard pattern over 12 runs of 96 samples. Correlation of the TM-ASR, Amplicor, and Versant assays was determined using 100 paired clinical samples and Deming regression analysis. The TM-ASR performed well with respect to linearity, precision, and contamination control. The correlation between TM-ASR and the Amplicor and Versant assays was poor, with large differences between assay results for individual samples. Calibration of the TM-ASR assay with Armored RNA allowed for a wide dynamic range and description of the distribution of HCV RNA in clinical samples.

  1. Evaluation of the COBAS Hepatitis C Virus (HCV) TaqMan Analyte-Specific Reagent Assay and Comparison to the COBAS Amplicor HCV Monitor V2.0 and Versant HCV bDNA 3.0 Assays

    PubMed Central

    Konnick, Eric Q.; Williams, Sheri M.; Ashwood, Edward R.; Hillyard, David R.

    2005-01-01

    Performance characteristics of the COBAS hepatitis C virus (HCV) TaqMan analyte-specific reagent (TM-ASR) assay using the QIAGEN BioRobot 9604 for RNA extraction were evaluated and compared to the COBAS Amplicor HCV Monitor V2.0 (Amplicor) and Versant HCV bDNA 3.0 (Versant) assays using clinical samples. Calibration of TM-ASR using Armored RNA allowed determination of the distribution of HCV RNA in clinical samples, using 22,399 clinical samples. Limit of detection, linearity, and inter- and intraassay assay precision were determined for the TM-ASR assay using multiple clinical specimen panels across multiple determinations. Genotype specificity for the TM-ASR assay was determined using samples with different HCV RNA genotypes evaluated and compared against predetermined results. Contamination control of the TM-ASR assay was evaluated using pools of HCV RNA-positive and -negative samples tested in a checkerboard pattern over 12 runs of 96 samples. Correlation of the TM-ASR, Amplicor, and Versant assays was determined using 100 paired clinical samples and Deming regression analysis. The TM-ASR performed well with respect to linearity, precision, and contamination control. The correlation between TM-ASR and the Amplicor and Versant assays was poor, with large differences between assay results for individual samples. Calibration of the TM-ASR assay with Armored RNA allowed for a wide dynamic range and description of the distribution of HCV RNA in clinical samples. PMID:15872232

  2. Effect of meal portion size choice on plate waste generation among patients with different nutritional status. An investigation using Dietary Intake Monitoring System (DIMS).

    PubMed

    Ofei, K T; Holst, M; Rasmussen, H H; Mikkelsen, B E

    2015-08-01

    The trolley meal system allows hospital patients to select food items and portion sizes directly from the food trolley. The nutritional status of the patient may be compromised if portions selected do not meet recommended intakes for energy, protein and micronutrients. The aim of this study was to investigate: (1) the portion size served, consumed and plate waste generated, (2) the extent to which the size of meal portions served contributes to daily recommended intakes for energy and protein, (3) the predictive effect of the served portion sizes on plate waste in patients screened for nutritional risk by NRS-2002, and (4) to establish the applicability of the dietary intake monitoring system (DIMS) as a technique to monitor plate waste. A prospective observational cohort study was conducted in two hospital wards over five weekdays. The DIMS was used to collect paired before- and after-meal consumption photos and measure the weight of plate content. The proportion of energy and protein consumed by both groups at each meal session could contribute up to 15% of the total daily recommended intake. Linear mixed model identified a positive relationship between meal portion size and plate waste (P = 0.002) and increased food waste in patients at nutritional risk during supper (P = 0.001). Meal portion size was associated with the level of plate waste produced. Being at nutritional risk further increased the extent of waste, regardless of the portion size served at supper. The use of DIMS as an innovative technique might be a promising way to monitor plate waste for optimizing meal portion size servings and minimizing food waste. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Fusion of microlitre water-in-oil droplets for simple, fast and green chemical assays.

    PubMed

    Chiu, S-H; Urban, P L

    2015-08-07

    A simple format for microscale chemical assays is proposed. It does not require the use of test tubes, microchips or microtiter plates. Microlitre-range (ca. 0.7-5.0 μL) aqueous droplets are generated by a commercial micropipette in a non-polar matrix inside a Petri dish. When two droplets are pipetted nearby, they spontaneously coalesce within seconds, priming a chemical reaction. Detection of the reaction product is accomplished by colorimetry, spectrophotometry, or fluorimetry using simple light-emitting diode (LED) arrays as the sources of monochromatic light, while chemiluminescence detection of the analytes present in single droplets is conducted in the dark. A smartphone camera is used as the detector. The limits of detection obtained for the developed in-droplet assays are estimated to be: 1.4 nmol (potassium permanganate by colorimetry), 1.4 pmol (fluorescein by fluorimetry), and 580 fmol (sodium hypochlorite by chemiluminescence detection). The format has successfully been used to monitor the progress of chemical and biochemical reactions over time with sub-second resolution. A semi-quantitative analysis of ascorbic acid using Tillman's reagent is presented. A few tens of individual droplets can be scanned in parallel. Rapid switching of the LED light sources with different wavelengths enables a spectral analysis of multiple droplets. Very little solid waste is produced. The assay matrix is readily recycled, thus the volume of liquid waste produced each time is also very small (typically, 1-10 μL per analysis). Various water-immiscible translucent liquids can be used as the reaction matrix: including silicone oil, 1-octanol as well as soybean cooking oil.

  4. A fully-integrated aptamer-based affinity assay platform for monitoring astronaut health in space.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yang, Xianbin; Durland, Ross H.; Hecht, Ariel H.

    2010-07-01

    Here we demonstrate the suitability of robust nucleic acid affinity reagents in an integrated point-of-care diagnostic platform for monitoring proteomic biomarkers indicative of astronaut health in spaceflight applications. A model thioaptamer targeting nuclear factor-kappa B (NF-{kappa}B) is evaluated in an on-chip electrophoretic gel-shift assay for human serum. Key steps of (i) mixing sample with the aptamer, (ii) buffer exchange, and (iii) preconcentration of sample were successfully integrated upstream of fluorescence-based detection. Challenges due to (i) nonspecific interactions with serum, and (ii) preconcentration at a nanoporous membrane are discussed and successfully resolved to yield a robust, rapid, and fully-integrated diagnostic system.

  5. A European multicientre study on the comparison of HBV viral loads between VERIS HBV assay and Roche COBAS® TAQMAN® HBV test, Abbott RealTime HBV assay, Siemens VERSANT HBV assay, and Qiagen artus HBV RG kit.

    PubMed

    Braun, Patrick; Delgado, Rafael; Drago, Monica; Fanti, Diana; Fleury, Hervé; Izopet, Jacques; Lombardi, Alessandra; Marcos, MaAngeles; Sauné, Karine; O'Shea, Siobhan; Pérez-Rivilla, Alfredo; Ramble, John; Trimoulet, Pascale; Vila, Jordi; Whittaker, Duncan; Artus, Alain; Rhodes, Daniel

    2017-10-01

    Hepatitis B viral load testing is essential to treatment and monitoring decisions in patients with chronic Hepatitis B. Beckman Coulter has developed the VERIS HBV Assay (Veris) for use on the fully automated DxN VERIS Molecular Diagnostics System. 1 OBJECTIVES: To evaluate the clinical performance of the Veris HBV Assay at multiple EU laboratories STUDY DESIGN: Method comparison was performed with a total of 344 plasma specimens from HBV infected patients tested with Veris and COBAS ® TaqMan ® HBV Test (Cobas), 207 specimens tested with Veris and RealTime HBV Assay (RealTime), 86 specimens tested with Veris and VERSANT ® HBV Assay (Versant), and 74 specimens tested with Veris and artus ® HBV RG PCR kit (artus). Bland-Altman analysis showed average bias of -0.46 log 10 IU/mL between Veris and Cobas, -0.46 log 10 IU/mL between Veris and RealTime, -0.36 log 10 IU/mL between Veris and Versant, and -0.12 log 10 IU/mL between Veris and artus. Bias was consistent across the assay range. Patient monitoring results using Veris demonstrated similar viral load trends over time to Cobas, RealTime, and artus. The VERIS HBV Assay demonstrated comparable clinical performance, with varying degrees of negative bias, compared to other currently marketed assays for HBV DNA monitoring. This negative bias should be taken into consideration if switching monitoring methods to Veris. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. A European multicientre study on the comparison of HIV-1 viral loads between VERIS HIV-1 Assay and Roche COBAS® TAQMAN® HIV-1 test, Abbott RealTime HIV-1 Assay, and Siemens VERSANT HIV-1 Assay.

    PubMed

    Braun, Patrick; Delgado, Rafael; Drago, Monica; Fanti, Diana; Fleury, Hervé; Hofmann, Jörg; Izopet, Jacques; Kühn, Sebastian; Lombardi, Alessandra; Mancon, Alessandro; Marcos, Mª Angeles; Mileto, Davide; Sauné, Karine; O'Shea, Siobhan; Pérez-Rivilla, Alfredo; Ramble, John; Trimoulet, Pascale; Vila, Jordi; Whittaker, Duncan; Artus, Alain; Rhodes, Daniel

    2017-07-01

    Viral load monitoring is essential for patients under treatment for HIV. Beckman Coulter has developed the VERIS HIV-1 Assay for use on the novel, automated DxN VERIS Molecular Diagnostics System. ¥ OBJECTIVES: Evaluation of the clinical performance of the new quantitative VERIS HIV-1 Assay at multiple EU laboratories. Method comparison with the VERIS HIV-1 Assay was performed with 415 specimens at 5 sites tested with COBAS ® AmpliPrep/COBAS ® TaqMan ® HIV-1 Test, v2.0, 169 specimens at 3 sites tested with RealTime HIV-1 Assay, and 202 specimens from 2 sites tested with VERSANT HIV-1 Assay. Patient monitoring sample results from 4 sites were also compared. Bland-Altman analysis showed the average bias between VERIS HIV-1 Assay and COBAS HIV-1 Test, RealTime HIV-1 Assay, and VERSANT HIV-1 Assay to be 0.28, 0.39, and 0.61 log 10 cp/mL, respectively. Bias at low end levels below 1000cp/mL showed predicted bias to be <0.3 log 10 cp/mL for VERIS HIV-1 Assay versus COBAS HIV-1 Test and RealTime HIV-1 Assay, and <0.5 log 10 cp/mL versus VERSANT HIV-1 Assay. Analysis on 174 specimens tested with the 0.175mL volume VERIS HIV-1 Assay and COBAS HIV-1 Test showed average bias of 0.39 log 10 cp/mL. Patient monitoring results using VERIS HIV-1 Assay demonstrated similar viral load trends over time to all comparators. The VERIS HIV-1 Assay for use on the DxN VERIS System demonstrated comparable clinical performance to COBAS ® HIV-1 Test, RealTime HIV-1 Assay, and VERSANT HIV-1 Assay. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. An optimized IFN-γ ELISpot assay for the sensitive and standardized monitoring of CMV protein-reactive effector cells of cell-mediated immunity.

    PubMed

    Barabas, Sascha; Spindler, Theresa; Kiener, Richard; Tonar, Charlotte; Lugner, Tamara; Batzilla, Julia; Bendfeldt, Hanna; Rascle, Anne; Asbach, Benedikt; Wagner, Ralf; Deml, Ludwig

    2017-03-07

    In healthy individuals, Cytomegalovirus (CMV) infection is efficiently controlled by CMV-specific cell-mediated immunity (CMI). Functional impairment of CMI in immunocompromized individuals however can lead to uncontrolled CMV replication and severe clinical complications. Close monitoring of CMV-specific CMI is therefore clinically relevant and might allow a reliable prognosis of CMV disease as well as assist personalized therapeutic decisions. Objective of this work was the optimization and technical validation of an IFN-γ ELISpot assay for a standardized, sensitive and reliable quantification of CMV-reactive effector cells. T-activated® immunodominant CMV IE-1 and pp65 proteins were used as stimulants. All basic assay parameters and reagents were tested and optimized to establish a user-friendly protocol and maximize the signal-to-noise ratio of the ELISpot assay. Optimized and standardized ELISpot revealed low intra-assay, inter-assay and inter-operator variability (coefficient of variation CV below 22%) and CV inter-site was lower than 40%. Good assay linearity was obtained between 6 × 10 4 and 2 × 10 5 PBMC per well upon stimulation with T-activated® IE-1 (R 2  = 0.97) and pp65 (R 2  = 0.99) antigens. Remarkably, stimulation of peripheral blood mononuclear cells (PBMC) with T-activated® IE-1 and pp65 proteins resulted in the activation of a broad range of CMV-reactive effector cells, including CD3 + CD4 + (Th), CD3 + CD8 + (CTL), CD3 - CD56 + (NK) and CD3 + CD56 + (NKT-like) cells. Accordingly, the optimized IFN-γ ELISpot assay revealed very high sensitivity (97%) in a cohort of 45 healthy donors, of which 32 were CMV IgG-seropositive. The combined use of T-activated® IE-1 and pp65 proteins for the stimulation of PBMC with the optimized IFN-γ ELISpot assay represents a highly standardized, valuable tool to monitor the functionality of CMV-specific CMI with great sensitivity and reliability.

  8. Measuring waste prevention.

    PubMed

    Zorpas, Antonis A; Lasaridi, Katia

    2013-05-01

    The Waste Framework Directive (WFD-2008/98/EC) has set clear waste prevention procedures, including reporting, reviewing, monitoring and evaluating. Based on the WFD, the European Commission and will offer support to Member States on how to develop waste prevention programmes through guidelines and information sharing on best practices. Monitoring and evaluating waste prevention activities are critical, as they constitute the main tools to enable policy makers, at the national and local level, to build their strategic plans and ensure that waste prevention initiatives are effective and deliver behaviour change. However, how one can measure something that is not there, remains an important and unresolved research question. The paper reviews and attempts to evaluate the methods that are being used for measuring waste prevention and the impact of relevant implemented activities at the household level, as the available data is still limited. Copyright © 2012 Elsevier Ltd. All rights reserved.

  9. Quantitative analysis of cytomegalovirus (CMV) viremia using the pp65 antigenemia assay and the COBAS AMPLICOR CMV MONITOR PCR test after blood and marrow allogeneic transplantation.

    PubMed

    Boivin, G; Bélanger, R; Delage, R; Béliveau, C; Demers, C; Goyette, N; Roy, J

    2000-12-01

    The performance of a commercially available qualitative PCR test for plasma (AMPLICOR CMV Test; Roche Diagnostics) and a quantitative PCR test for plasma and leukocytes (COBAS AMPLICOR CMV MONITOR Test; Roche Diagnostics) was evaluated with samples from 50 blood or marrow allogeneic transplant recipients who received short courses of sequential ganciclovir therapy (2 weeks intravenously followed by 2 weeks orally) based on a positive cytomegalovirus (CMV) pp65 antigenemia (AG) assay. The number of persons with a positive CMV test was significantly higher for leukocyte-based assays (AG, 67.5%; PCR, 62.5%) compared to both quantitative and qualitative PCR tests of plasma (42.5 and 35%, respectively). One person developed CMV disease during the study despite a negative AG assay; in this particular case, all PCR assays were found to be positive 10 days before his death. There was a trend for earlier positivity after transplantation and more rapid negativity after initiation of ganciclovir for the tests performed on leukocytes. The mean number of CMV copies as assessed by PCR was significantly higher in leukocytes than in plasma (P = 0.02). Overall, excellent agreement (kappa coefficient, >0.75) was found only between the two PCR assays (qualitative and quantitative) based on plasma. These results suggest that either the pp65 AG assay or the COBAS AMPLICOR CMV MONITOR Test using leukocytes could be used to safely monitor CMV viremia in related allogeneic blood or marrow transplant recipients. Such a strategy will result in preemptive treatment for about two-thirds of the persons with a relatively low rate (<33%) of secondary viremic episodes following short courses of ganciclovir therapy.

  10. Generic HPLC platform for automated enzyme reaction monitoring: Advancing the assay toolbox for transaminases and other PLP-dependent enzymes.

    PubMed

    Börner, Tim; Grey, Carl; Adlercreutz, Patrick

    2016-08-01

    Methods for rapid and direct quantification of enzyme kinetics independent of the substrate stand in high demand for both fundamental research and bioprocess development. This study addresses the need for a generic method by developing an automated, standardizable HPLC platform monitoring reaction progress in near real-time. The method was applied to amine transaminase (ATA) catalyzed reactions intensifying process development for chiral amine synthesis. Autosampler-assisted pipetting facilitates integrated mixing and sampling under controlled temperature. Crude enzyme formulations in high and low substrate concentrations can be employed. Sequential, small (1 µL) sample injections and immediate detection after separation permits fast reaction monitoring with excellent sensitivity, accuracy and reproducibility. Due to its modular design, different chromatographic techniques, e.g. reverse phase and size exclusion chromatography (SEC) can be employed. A novel assay for pyridoxal 5'-phosphate-dependent enzymes is presented using SEC for direct monitoring of enzyme-bound and free reaction intermediates. Time-resolved changes of the different cofactor states, e.g. pyridoxal 5'-phosphate, pyridoxamine 5'-phosphate and the internal aldimine were traced in both half reactions. The combination of the automated HPLC platform with SEC offers a method for substrate-independent screening, which renders a missing piece in the assay and screening toolbox for ATAs and other PLP-dependent enzymes. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Radiological Monitoring Results for Groundwater Samples Associated with the Industrial Wastewater Reuse Permit for the Materials and Fuels Complex Industrial Waste Ditch and Pond: November 1, 2011-October 31, 2012

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mike lewis

    2013-02-01

    This report summarizes radiological monitoring performed on samples from specific groundwater monitoring wells associated with the Industrial Wastewater Reuse Permit for the Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond WRU-I-0160-01, Modification 1 (formerly LA-000160-01). The radiological monitoring was performed to fulfill Department of Energy requirements under the Atomic Energy Act.

  12. Radiological Monitoring Results for Groundwater Samples Associated with the Industrial Wastewater Reuse Permit for the Materials and Fuels Complex Industrial Waste Ditch and Pond: November 1, 2012-October 31, 2013

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mike Lewis

    2014-02-01

    This report summarizes radiological monitoring performed on samples from specific groundwater monitoring wells associated with the Industrial Wastewater Reuse Permit for the Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond WRU-I-0160-01, Modification 1 (formerly LA-000160-01). The radiological monitoring was performed to fulfill Department of Energy requirements under the Atomic Energy Act.

  13. Traditional and Model Based Assay of Irregular Geometry Items

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    MOORE, FRANK S.; SALAYMEH, SALEEM

    The Analytical Development Section (ADS) of SRNL was requested to perform a waste disposal assay of two heater boxes which had been used in the HB Line dissolvers. They had been sent to SRNL for study to make recommendations on how to prevent future failure of the units when they were replaced. The study having been completed, the units needed to be characterized prior to sending to Solid Waste for disposal. An assay station consisting of a turntable, HPGe detector, CANBERRA Inspector, transmission source and a portable computer was set up to do the required assays. The assays indicate themore » presence of U-235, Pu-239 and Cs-137. No measurable amounts of U-235 or Pu-239 were found. Therefore the Minimum Detectable Activities for U-235 and Pu-239 were calculated. For Heater Box 1, 0.23 grams of U-235 and 0.24 grams of Pu-239. For Heater Box 2, the results were 0.21 grams of U-235 and 0.21 grams of Pu-239. This paper describes and documents the assays employed to determine the amount of U, Pu and Cs contents of the heater boxes. The paper provides results of SNM assays using traditional calibration of the system and on one based on modeling. It also provides the scientific community with data that will assist the user in determining the method of choice for assaying items with irregular geometries.« less

  14. Monitoring Ligand-Activated Protein-Protein Interactions Using Bioluminescent Resonance Energy Transfer (BRET) Assay.

    PubMed

    Coriano, Carlos; Powell, Emily; Xu, Wei

    2016-01-01

    The bioluminescent resonance energy transfer (BRET) assay has been extensively used in cell-based and in vivo imaging systems for detecting protein-protein interactions in the native environment of living cells. These protein-protein interactions are essential for the functional response of many signaling pathways to environmental chemicals. BRET has been used as a toxicological tool for identifying chemicals that either induce or inhibit these protein-protein interactions. This chapter focuses on describing the toxicological applications of BRET and its optimization as a high-throughput detection system in live cells. Here we review the construction of BRET fusion proteins, describe the BRET methodology, and outline strategies to overcome obstacles that may arise. Furthermore, we describe the advantage of BRET over other resonance energy transfer methods for monitoring protein-protein interactions.

  15. Monitoring estuarine circulation and ocean waste dispersion using an integrated satellite-aircraft-drogue approach. [Continental Shelf and Delaware Bay

    NASA Technical Reports Server (NTRS)

    Klemas, V. (Principal Investigator); Davis, G. R.; Wang, H.

    1975-01-01

    The author has identified the following significant results. An integrated satellite-aircraft-drogue approach was developed which employs remotely tracked expendable drogues together with satellite and aircraft observations of oil slicks, waste plumes, and natural tracers, such as suspended sediment. Tests conducted on the Continental Shelf and in Delaware Bay indicate that the system provides a cost effective means of monitoring current circulation and verifying oil slick and ocean waste dispersion models even under severe environmental conditions.

  16. Multilaboratory comparison of hepatitis C virus viral load assays.

    PubMed

    Caliendo, A M; Valsamakis, A; Zhou, Y; Yen-Lieberman, B; Andersen, J; Young, S; Ferreira-Gonzalez, A; Tsongalis, G J; Pyles, R; Bremer, J W; Lurain, N S

    2006-05-01

    We report a multilaboratory evaluation of hepatitis C virus (HCV) viral load assays to determine their linear range, reproducibility, subtype detection, and agreement. A panel of HCV RNA samples ranging in nominal concentration from 1.0 to 7.0 log10 IU/ml was constructed by diluting a clinical specimen (genotype 1b). Replicates of the panel were tested in multiple laboratories using the Abbott TaqMan analyte-specific reagent (Abbott reverse transcription-PCR [RT-PCR]), Roche TaqMan RUO (Roche RT-PCR), Roche Amplicor Monitor HCV 2.0 (Roche Monitor), and Bayer VERSANT HCV RNA 3.0 (Bayer bDNA) assays. Bayer bDNA-negative specimens were tested reflexively using the Bayer VERSANT HCV RNA qualitative assay (Bayer TMA). Abbott RT-PCR and Roche RT-PCR detected all 28 replicates with a concentration of 1.0 log10 IU/ml and were linear to 7.0 log10 IU/ml. Roche Monitor and Bayer bDNA detected 27 out of 28 and 13 out of 28 replicates, respectively, of 3.0 log10 IU/ml. Bayer TMA detected all seven replicates with 1.0 log10 IU/ml. Bayer bDNA was the most reproducible of the four assays. The mean viral load values for panel members in the linear ranges of the assays were within 0.5 log10 for the different tests. Eighty-nine clinical specimens of various genotypes (1 through 4) were tested in the Bayer bDNA, Abbott RT-PCR, and Roche RT-PCR assays. For Abbott RT-PCR, mean viral load values were 0.61 to 0.96 log10 greater than the values for Bayer bDNA assay for samples with genotype 1, 2, or 3 samples and 0.08 log10 greater for genotype 4 specimens. The Roche RT-PCR assay gave mean viral load values that were 0.28 to 0.82 log10 greater than those obtained with the Bayer bDNA assay for genotype 1, 2, and 3 samples. However, for genotype 4 samples the mean viral load value obtained with the Roche RT-PCR assay was, on average, 0.15 log10 lower than that of the Bayer bDNA. Based on these data, we conclude that the sensitivity and linear range of the Abbott and Roche RT-PCR assays

  17. Nevada National Security Site 2011 Waste Management Monitoring Report, Area 3 and Area 5 Radioactive Waste Management Site

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    NSTec Environmental Management

    2012-07-31

    Environmental monitoring data are collected at and around the Area 3 and Area 5 Radioactive Waste Management Sites (RWMSs) at the Nevada National Security Site (NNSS). These data are associated with radiation exposure, air, groundwater, meteorology, and vadose zone. This report summarizes the 2011 environmental data to provide an overall evaluation of RWMS performance and to support environmental compliance and performance assessment (PA) activities. Some of these data (e.g., radiation exposure, air, and groundwater) are presented in other reports. Direct radiation monitoring data indicate exposure levels at the RWMSs are within the range of background levels measured at the NNSS.more » Slightly elevated exposure levels outside the Area 3 RWMS are attributed to nearby historical aboveground nuclear weapons tests. Air monitoring data show tritium concentrations in water vapor and americium and plutonium concentrations in air particles are only slightly above detection limits and background levels. The measured levels of radionuclides in air particulates and moisture are below derived concentration guides for these radionuclides. During the last 2 weeks of March 2011, gamma spectroscopy results for air particles showed measurable activities of iodine-131 (131I), cesium-134 (134Cs), and cesium-137 (137Cs). These results are attributed to the release of fission products from the damaged Fukushima Daiichi power plant in Japan. The remaining gamma spectroscopy results for air particulates collected at the Area 3 and Area 5 RWMS were below minimum detectable concentrations. Groundwater monitoring data indicate the groundwater in the uppermost aquifer beneath the Area 5 RWMS is not impacted by RWMS operations. Results of groundwater analysis from wells around the Area 5 RWMS were all below established investigation levels. The 86.3 millimeters (mm) (3.40 inches [in.]) of precipitation at the Area 3 RWMS during 2011 is 44% below the average of 154.1 mm (6.07 in.), and the

  18. Tolerance assays: measuring the unknown.

    PubMed

    Newell, Kenneth A; Larsen, Christian P

    2006-06-15

    Distinguishing transplant recipients who will benefit from a reduction in, or even the withdrawal of, immunosuppression from those who require intensive, lifelong immunosuppression will be dependent on developing strategies for immune monitoring. Currently, no assays have been shown to accurately predict the development or presence of donor-specific tolerance in humans after transplantation. In this overview we describe and discuss those assays that we believe may be useful for identifying tolerant transplant recipients. Validation of "tolerance" assays will be critical for the safe development of tolerance regimens in humans.

  19. Monitoring regulation of DNA repair activities of cultured cells in-gel using the comet assay

    PubMed Central

    Nickson, Catherine M.; Parsons, Jason L.

    2014-01-01

    Base excision repair (BER) is the predominant cellular mechanism by which human cells repair DNA base damage, sites of base loss, and DNA single strand breaks of various complexity, that are generated in their thousands in every human cell per day as a consequence of cellular metabolism and exogenous agents, including ionizing radiation. Over the last three decades the comet assay has been employed in scientific research to examine the cellular response to these types of DNA damage in cultured cells, therefore revealing the efficiency and capacity of BER. We have recently pioneered new research demonstrating an important role for post-translational modifications (particularly ubiquitylation) in the regulation of cellular levels of BER proteins, and that subtle changes (∼20–50%) in protein levels following siRNA knockdown of E3 ubiquitin ligases or deubiquitylation enzymes can manifest in significant changes in DNA repair capacity monitored using the comet assay. For example, we have shown that the E3 ubiquitin ligase Mule, the tumor suppressor protein ARF, and the deubiquitylation enzyme USP47 modulate DNA repair by controlling cellular levels of DNA polymerase β, and also that polynucleotide kinase phosphatase levels are controlled by ATM-dependant phosphorylation and Cul4A–DDB1–STRAP-dependent ubiquitylation. In these studies we employed a modification of the comet assay whereby cultured cells, following DNA damage treatment, are embedded in agarose and allowed to repair in-gel prior to lysis and electrophoresis. Whilst this method does have its limitations, it avoids the extensive cell culture-based processing associated with the traditional approach using attached cells and also allows for the examination of much more precise DNA repair kinetics. In this review we will describe, using this modified comet assay, our accumulating evidence that ubiquitylation-dependant regulation of BER proteins has important consequences for overall cellular DNA repair

  20. Digital Automation and Real-Time Monitoring of an Original Installation for "Wet Combustion" of Organic Wastes

    NASA Astrophysics Data System (ADS)

    Morozov, Yegor; Tikhomirov, Alexander A.; Saltykov, Mikhail; Trifonov, Sergey V.; Kudenko, D.. Yurii A.

    2016-07-01

    An original method for "wet combustion" of organic wastes, which is being developed at the IBP SB RAS, is a very promising approach for regeneration of nutrient solutions for plants in future spacecraft closed Bioregenerative Life Support Systems (BLSS). The method is quick, ecofriendly, does not require special conditions such as high pressure and temperature, and the resulting nitrogen stays in forms easy for further preparation of the fertilizer. An experimental testbed of a new-generation closed ecosystem is being currently run at the IBP SB RAS to examine compatibility of the latest technologies for accelerating the cycling. Integration of "wet combustion" of organic wastes into the information system of closed ecosystem experimental testbed has been studied as part of preparatory work. Digital automation and real-time monitoring of original "wet combustion" installation operation parameters have been implemented. The new system enabled remotely controlled or automatic work of the installation. Data are stored in standard easily processed formats, allowing further mathematical processing where necessary. During ongoing experiments on improving "wet combustion" of organic wastes, automatic monitoring can notice slight changes in process parameters and record them in more detail. The ultimate goal of the study is to include the "wet combustion" installation into future full-scale experiment with humans, thus reducing the time spent by the crew on life support issues while living in the BLSS. The work was carried out with the financial support of the Russian Scientific Foundation (project 14-14-00599).

  1. Clinical performances of two real-time PCR assays and bDNA/TMA to early monitor treatment outcome in patients with chronic hepatitis C.

    PubMed

    Martinot-Peignoux, Michelle; Khiri, Hacène; Leclere, Laurence; Maylin, Sarah; Marcellin, Patrick; Halfon, Philippe

    2009-11-01

    Early viral monitoring is essential for the management of treatment outcome in patients with chronic hepatitis C. A variety of commercially available assays are now available to quantify HCV-RNA in routine clinical practice. Compare the clinical results of 3 commercially available assays to evaluate the positive predictive value (PPV) and the negative predictive value (NPV) of rapid virological response (RVR) at week 4 and early virological response (EVR) at week 12. 287 patients treated with standard care regimen combination therapy were studied. HCV-RNA values measured at baseline, week 4, week 12 with VERSANT HCV 3.0 Assay (bDNA), and VERSANT HCV-RNA Qualitative Assay (TMA) (bDNA/TMA); COBAS Ampliprep/COBAS/TaqMan (CAP/CTM) and Abbott m2000sp extraction/m2000rt amplification system (ART). RVR was defined as undetectable serum HCV-RNA and EVR as a > OR =2 log decline in baseline viral load (BLV). Median (range) BVLs were: 5.585(2.585-6.816), 5.189(2.792-7.747) and 4.804(2.380-6.580) log(10)IU/ml, with bDNA/TMA, CAP/CTM and ART, respectively (p<0.01); RVR was observed in 22%, 30% and 27% of the patients and PPVs were 97%, 91% and 94% with bDNA/TMA, CAP/CTM and ART, respectively (p=0.317). EVR was observed in 76%, 73% and 67% of the patients and NPVs were 93%, 83% and 79% with bDNA/TMA, CAP/CTM and ART, respectively (p=0.09). Treatment monitoring should include both detection of serum HCV-RNA at week 4 to predict SVR and at week 12 to predict non-SVR. The value of all 3 assays was similar for evaluating RVR or EVR. Because of viral load discrepancies the same assay should be used throughout patient treatment follow-up.

  2. Understanding the role of waste prevention in local waste management: A literature review.

    PubMed

    Zacho, Kristina O; Mosgaard, Mette A

    2016-10-01

    Local waste management has so far been characterised by end-of-pipe solutions, landfilling, incineration, and recycling. End-of-pipe solutions build on a different mind-set than life cycle-based approaches, and for this reason, local waste managers are reluctant to consider strategies for waste prevention. To accelerate the transition of waste and resource management towards a more integrated management, waste prevention needs to play a larger role in the local waste management. In this review article, we collect knowledge from the scientific community on waste prevention of relevance to local waste management. We analyse the trends in the waste prevention literature by organising the literature into four categories. The results indicate an increasing interest in waste prevention, but not much literature specifically concerns the integration of prevention into the local waste management. However, evidence from the literature can inform local waste management on the prevention potential; the environmental and social effects of prevention; how individuals in households can be motivated to reduce waste; and how the effects of prevention measures can be monitored. Nevertheless, knowledge is still lacking on local waste prevention, especially regarding the methods for monitoring and how local waste management systems can be designed to encourage waste reduction in the households. We end the article with recommendations for future research. The literature review can be useful for both practitioners in the waste sector and for academics seeking an overview of previous research on waste prevention. © The Author(s) 2016.

  3. NRC Waste Incidental to Reprocessing Program: Overview of Consultation and Monitoring Activities at the Idaho National Laboratory and the Savannah River Site - What We Have Learned - 12470

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Suber, Gregory

    2012-07-01

    In 2005 the U.S. Nuclear Regulatory Commission (NRC) began to implement a new set of responsibilities under the Ronald W. Reagan National Defense Authorization Act (NDAA) of Fiscal Year 2005. Section 3116 of the NDAA requires the U.S. Department of Energy (DOE) to consult with the NRC for certain non-high level waste determinations and also requires NRC to monitor DOE's disposal actions related to those determinations. In Fiscal Year 2005, the NRC staff began consulting with DOE and completed reviews of draft waste determinations for salt waste at the Savannah River Site. In 2006, a second review was completed onmore » tank waste residuals including sodium-bearing waste at the Idaho Nuclear Technology and Engineering Center Tank Farm at the Idaho National Laboratory. Monitoring Plans were developed for these activities and the NRC is actively monitoring disposal actions at both sites. NRC is currently in consultation with DOE on the F-Area Tank Farm closure and anticipates entering consultation on the H-Area Tank Farm at the Savannah River Site. This paper presents, from the NRC perspective, an overview of how the consultation and monitoring process has evolved since its conception in 2005. It addresses changes in methods and procedures used to collect and develop information used by the NRC in developing the technical evaluation report and monitoring plan under consultation and the implementation the plan under monitoring. It will address lessons learned and best practices developed throughout the process. The NDAA has presented significant challenges for the NRC and DOE. Past and current successes demonstrate that the NDAA can achieve its intended goal of facilitating tank closure at DOE legacy defense waste sites. The NRC believes many of the challenges in performing the WD reviews have been identified and addressed. Lessons learned have been collected and documented throughout the review process. Future success will be contingent on each agencies commitment

  4. Cytomegalovirus (CMV) immune monitoring with ELISPOT and QuantiFERON-CMV assay in seropositive kidney transplant recipients

    PubMed Central

    Ryu, Ji Hyeong; Choi, Ae-Ran; Yu, Ji Hyun; Lim, Jihyang; Han, Kyungja; Kim, Sang Il; Yang, Chul Woo; Chung, Byung Ha

    2017-01-01

    Although cytomegalovirus (CMV) specific cell-mediated immunity (CMI) has been suggested as a predictive marker for CMV infection, proper CMI monitoring strategy in CMV-seropositive recipients and optimal method are not defined. The aim of this study was to evaluate two interferon gamma release assays during early post-transplant period as a predictor of the development of CMV infection in CMV-seropositive patients. A total of 124 CMV-seropositive recipients who received kidney transplantation from CMV-seropositive donor were prospectively examined. At pre-transplant and post-transplant 1 and 3 months, CMV-CMIs were tested using QuantiFERON-CMV assay (QF-CMV) and CMV specific T cell ELISPOT against CMV pp65 and IE-1 antigens (pp65-ELISPOT, IE-1-ELISPOT). CMV DNAemia occurred in 16 (12.9%) patients within 3 months after transplant. Post-transplant pp65 or IE-1 ELISPOT response, but not QF-CMV, was significantly associated with CMV DNAemia. The pp65 ELISPOT (cut-off; 30 spots/200,000 cells) and IE-1 ELISPOT (10 spots/200,000 cells) at post-transplant 1 month predicted the risk of post-transplant CMV DNAemia (P = 0.019). Negative predictive values (NPV) for protection from CMV DNAemia in case of positive ELISPOT results were 94.5% (95% CI: 86.9–97.8%) and 97.6% (95% CI: 86.3–99.6%) in pp65-ELISPOT and IE-1-ELISPOT assays, respectively. These results suggest that the variability may exist between CMV ELISPOT assays and QF-CMV, and CMV ELISPOT at post-transplant 1 month can identify the risk of CMV DNAemia in seropositive kidney transplant recipients. PMID:29232714

  5. Cytomegalovirus (CMV) immune monitoring with ELISPOT and QuantiFERON-CMV assay in seropositive kidney transplant recipients.

    PubMed

    Lee, Hyeyoung; Park, Ki Hyun; Ryu, Ji Hyeong; Choi, Ae-Ran; Yu, Ji Hyun; Lim, Jihyang; Han, Kyungja; Kim, Sang Il; Yang, Chul Woo; Chung, Byung Ha; Oh, Eun-Jee

    2017-01-01

    Although cytomegalovirus (CMV) specific cell-mediated immunity (CMI) has been suggested as a predictive marker for CMV infection, proper CMI monitoring strategy in CMV-seropositive recipients and optimal method are not defined. The aim of this study was to evaluate two interferon gamma release assays during early post-transplant period as a predictor of the development of CMV infection in CMV-seropositive patients. A total of 124 CMV-seropositive recipients who received kidney transplantation from CMV-seropositive donor were prospectively examined. At pre-transplant and post-transplant 1 and 3 months, CMV-CMIs were tested using QuantiFERON-CMV assay (QF-CMV) and CMV specific T cell ELISPOT against CMV pp65 and IE-1 antigens (pp65-ELISPOT, IE-1-ELISPOT). CMV DNAemia occurred in 16 (12.9%) patients within 3 months after transplant. Post-transplant pp65 or IE-1 ELISPOT response, but not QF-CMV, was significantly associated with CMV DNAemia. The pp65 ELISPOT (cut-off; 30 spots/200,000 cells) and IE-1 ELISPOT (10 spots/200,000 cells) at post-transplant 1 month predicted the risk of post-transplant CMV DNAemia (P = 0.019). Negative predictive values (NPV) for protection from CMV DNAemia in case of positive ELISPOT results were 94.5% (95% CI: 86.9-97.8%) and 97.6% (95% CI: 86.3-99.6%) in pp65-ELISPOT and IE-1-ELISPOT assays, respectively. These results suggest that the variability may exist between CMV ELISPOT assays and QF-CMV, and CMV ELISPOT at post-transplant 1 month can identify the risk of CMV DNAemia in seropositive kidney transplant recipients.

  6. Remote Sensing Combined with Field Spectroscopy for the Detection and Monitoring of Heavy Metal Contamination from Informal E-waste Recycling

    NASA Astrophysics Data System (ADS)

    Friedlander, L. R.; Garb, Y.

    2017-12-01

    Electronic waste (e-waste) is one of today's fastest growing waste streams. Made up of discarded electronics, e-waste disposal is complex. However, e-waste also provides economic opportunity through the processing and extraction of precious metals. Sometimes referred to as "urban mining," this recycling operates informally or illegally and is characterized by dangerous practices such as, open-pit burning, acid leaching, and burning of low value wastes. Poorly controlled e-waste recycling releases dangerous contaminants, especially heavy metals, directly to the surface environment where they can infiltrate water resources and spread through precipitation events. Despite growing recognition of the prevalence of unregulated e-waste processing, systematic data on the extent and persistence of the released contamination is still limited. In general, contamination is established through techniques that provide only a snapshot in time and in a limited geographic area. Here we present preliminary results from attempts to combine field, laboratory, and remote sensing studies toward a systematic remote sensing methodology for e-waste contamination detection and monitoring. The ongoing work utilizes a tragic "natural experiment," in which over 500 e-waste burn sites were active over more than a decade in a variety of agricultural, residential, and natural contexts. We have collected over 100 soil samples for which we have both XRF and ICP-AES measurements showing soil Pb concentrations as high as 14000 ppm. We have also collected 480 in-situ reflectance spectra with corresponding soil samples over 4 field transects of areas with long-term burn activity. The most heavily contaminated samples come from within the burn sites and are made up of ash. Field spectra of these samples reflect their dark color with low overall reflectance and shallow spectral features. These spectra are challenging to use for image classification due to their similarity with other low-reflectance parts

  7. Multiplex Liquid Chromatography-Tandem Mass Spectrometry Assay for Simultaneous Therapeutic Drug Monitoring of Ribavirin, Boceprevir, and Telaprevir

    PubMed Central

    Aouri, Manel; Moradpour, Darius; Cavassini, Matthias; Mercier, Thomas; Buclin, Thierry; Csajka, Chantal; Telenti, Amalio; Rauch, Andri

    2013-01-01

    New directly acting antivirals (DAAs) that inhibit hepatitis C virus (HCV) replication are increasingly used for the treatment of chronic hepatitis C. A marked pharmacokinetic variability and a high potential for drug-drug interactions between DAAs and numerous drug classes have been identified. In addition, ribavirin (RBV), commonly associated with hemolytic anemia, often requires dose adjustment, advocating for therapeutic drug monitoring (TDM) in patients under combined antiviral therapy. However, an assay for the simultaneous analysis of RBV and DAAs constitutes an analytical challenge because of the large differences in polarity among these drugs, ranging from hydrophilic (RBV) to highly lipophilic (telaprevir [TVR]). Moreover, TVR is characterized by erratic behavior on standard octadecyl-based reversed-phase column chromatography and must be separated from VRT-127394, its inactive C-21 epimer metabolite. We have developed a convenient assay employing simple plasma protein precipitation, followed by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) for the simultaneous determination of levels of RBV, boceprevir, and TVR, as well as its metabolite VRT-127394, in plasma. This new, simple, rapid, and robust HPLC-MS/MS assay offers an efficient method of real-time TDM aimed at maximizing efficacy while minimizing the toxicity of antiviral therapy. PMID:23629707

  8. WASTE TREATMENT BUILDING SYSTEM DESCRIPTION DOCUMENT

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    F. Habashi

    2000-06-22

    The Waste Treatment Building System provides the space, layout, structures, and embedded subsystems that support the processing of low-level liquid and solid radioactive waste generated within the Monitored Geologic Repository (MGR). The activities conducted in the Waste Treatment Building include sorting, volume reduction, and packaging of dry waste, and collecting, processing, solidification, and packaging of liquid waste. The Waste Treatment Building System is located on the surface within the protected area of the MGR. The Waste Treatment Building System helps maintain a suitable environment for the waste processing and protects the systems within the Waste Treatment Building (WTB) from mostmore » of the natural and induced environments. The WTB also confines contaminants and provides radiological protection to personnel. In addition to the waste processing operations, the Waste Treatment Building System provides space and layout for staging of packaged waste for shipment, industrial and radiological safety systems, control and monitoring of operations, safeguards and security systems, and fire protection, ventilation and utilities systems. The Waste Treatment Building System also provides the required space and layout for maintenance activities, tool storage, and administrative facilities. The Waste Treatment Building System integrates waste processing systems within its protective structure to support the throughput rates established for the MGR. The Waste Treatment Building System also provides shielding, layout, and other design features to help limit personnel radiation exposures to levels which are as low as is reasonably achievable (ALARA). The Waste Treatment Building System interfaces with the Site Generated Radiological Waste Handling System, and with other MGR systems that support the waste processing operations. The Waste Treatment Building System interfaces with the General Site Transportation System, Site Communications System, Site Water System

  9. Active and passive computed tomography mixed waste focus area final report

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Roberson, G P

    1998-08-19

    The Mixed Waste Focus Area (MWFA) Characterization Development Strategy delineates an approach to resolve technology deficiencies associated with the characterization of mixed wastes. The intent of this strategy is to ensure the availability of technologies to support the Department of Energy's (DOE) mixed waste low-level or transuranic (TRU) contaminated waste characterization management needs. To this end the MWFA has defined and coordinated characterization development programs to ensure that data and test results necessary to evaluate the utility of non-destructive assay technologies are available to meet site contact handled waste management schedules. Requirements used as technology development project benchmarks are basedmore » in the National TRU Program Quality Assurance Program Plan. These requirements include the ability to determine total bias and total measurement uncertainty. These parameters must be completely evaluated for waste types to be processed through a given nondestructive waste assay system constituting the foundation of activities undertaken in technology development projects. Once development and testing activities have been completed, Innovative Technology Summary Reports are generated to provide results and conclusions to support EM-30, -40, or -60 end user/customer technology selection. The Active and Passive Computed Tomography non-destructive assay system is one of the technologies selected for development by the MWFA. Lawrence Livermore National Laboratory's (LLNL) is developing the Active and Passive Computed Tomography (A&PCT) nondestructive assay (NDA) technology to identify and accurately quantify all detectable radioisotopes in closed containers of waste. This technology will be applicable to all types of waste regardless of .their classification; low level, transuranic or provide results and conclusions to support EM-30, -40, or -60 end user/customer technology selection. The Active and Passive Computed Tomography non

  10. Evaluation Model of Plate Waste to Monitor Food Consumption in Two Different Catering Settings

    PubMed Central

    Scognamiglio, Umberto; Moroni, Catia; Marani, Alessandra; Calcaterra, Veronica; Amendola, Mariano; Civitelli, Giulia; Cattaruzza, Maria Sofia; Ermenegildi, Arianna; Morena, Valeria

    2014-01-01

    An increasing number of people regularly eats lunch away from home, using catering services. In this context, therefore, it is extremely important to improve the meals’ quality, remaining faithful to the principles of hygiene, nutritional and organoleptic quality and proper food handling. At the same time, it is necessary to promote food choices, nutritionally correct, by evaluations of appropriateness of menus. The study of food waste allows an evaluation of the nutritional habits of consumers and an important economic consideration of the costs incurred for the implementation of the service. This becomes even more important in some particularly sensitive groups, such as children and elderly. The purpose of this work is to test a model of semi-quantitative evaluation of waste to monitor food consumption in two different catering contexts (educational and business), in order to improve the service for school students and other consumers. PMID:27800337

  11. Evaluation Model of Plate Waste to Monitor Food Consumption in Two Different Catering Settings.

    PubMed

    Saccares, Stefano; Scognamiglio, Umberto; Moroni, Catia; Marani, Alessandra; Calcaterra, Veronica; Amendola, Mariano; Civitelli, Giulia; Cattaruzza, Maria Sofia; Ermenegildi, Arianna; Morena, Valeria

    2014-04-17

    An increasing number of people regularly eats lunch away from home, using catering services. In this context, therefore, it is extremely important to improve the meals' quality, remaining faithful to the principles of hygiene, nutritional and organoleptic quality and proper food handling. At the same time, it is necessary to promote food choices, nutritionally correct, by evaluations of appropriateness of menus. The study of food waste allows an evaluation of the nutritional habits of consumers and an important economic consideration of the costs incurred for the implementation of the service. This becomes even more important in some particularly sensitive groups, such as children and elderly. The purpose of this work is to test a model of semi-quantitative evaluation of waste to monitor food consumption in two different catering contexts (educational and business), in order to improve the service for school students and other consumers.

  12. INNOVATIVE CONCEPTS FOR DETECTING AND LOCATING LEAKS IN WASTE IMPOUNDMENT LINER SYSTEMS: ACOUSTIC EMISSION MONITORING AND TIME DOMAIN REFLECTOMETRY

    EPA Science Inventory

    This project is part of a program to investigate the use of innovative techniques for detecting and locating leaks in waste impoundment liners. Laboratory and small scale field studies were undertaken to evaluate the potential of Acoustic Emission Monitoring (AEM) and Time Domain...

  13. Environment Monitor

    NASA Technical Reports Server (NTRS)

    1988-01-01

    Viking landers touched down on Mars equipped with a variety of systems to conduct automated research, each carrying a compact but highly sophisticated instrument for analyzing Martian soil and atmosphere. Instrument called a Gas Chromatography/Mass Spectrometer (GC/MS) had to be small, lightweight, shock resistant, highly automated and extremely sensitive, yet require minimal electrical power. Viking Instruments Corporation commercialized this technology and targeted their primary market as environmental monitoring, especially toxic and hazardous waste site monitoring. Waste sites often contain chemicals in complex mixtures, and the conventional method of site characterization, taking samples on-site and sending them to a laboratory for analysis is time consuming and expensive. Other terrestrial applications are explosive detection in airports, drug detection, industrial air monitoring, medical metabolic monitoring and for military, chemical warfare agents.

  14. Implementation of spatial smart waste management system in malaysia

    NASA Astrophysics Data System (ADS)

    Omar, M. F.; Termizi, A. A. A.; Zainal, D.; Wahap, N. A.; Ismail, N. M.; Ahmad, N.

    2016-06-01

    One of the challenges to innovate and create an IoT -enabled solution is in monitoring and management of the environment. Waste collection utilizing the Internet of Things (IoT) with the technology of smart wireless sensors will able to gather fill-level data from waste containers hence providing a waste monitoring solution that brings up savings in waste collection costs. One of the challenges to the local authority is how to monitor the works of contractor effective and efficiently in waste management. This paper will propose to the local authority the implementation of smart waste management in Malaysia to improve the city management and to provide better services to the public towards smart city applications.

  15. DNA aptamer beacon assay for C-telopeptide and handheld fluorometer to monitor bone resorption.

    PubMed

    Bruno, John Gordon; Carrillo, Maria P; Phillips, Taylor; Hanson, Douglas; Bohmann, Jonathan A

    2011-09-01

    A novel DNA aptamer beacon is described for quantification of a 26-amino acid C-telopeptide (CTx) of human type I bone collagen. One aptamer sequence and its reverse complement dominated the aptamer pool (31.6% of sequenced clones). Secondary structures of these aptamers were examined for potential binding pockets. Three-dimensional computer models which analyzed docking topologies and binding energies were in agreement with empirical fluorescence experiments used to select one candidate loop for beacon assay development. All loop structures from the aptamer finalists were end-labeled with TYE 665 and Iowa Black quencher for comparison of beacon fluorescence levels as a function of CTx concentration. The optimal beacon, designated CTx 2R-2h yielded a low ng/ml limit of detection using a commercially available handheld fluorometer. The CTx aptamer beacon bound full-length 26-amino acid CTx peptide, but not a shorter 8-amino acid segment of CTx peptide which is a common target for commercial CTx ELISA kits. The prototype assay was shown to detect CTx peptide from human urine after creatinine and urea were removed by size-exclusion chromatography to prevent nonspecific denaturing of the aptamer beacon. This work demonstrates the potential of aptamer beacons to be utilized for rapid and sensitive bone health monitoring in a handheld or point-of-care format.

  16. Development and evaluation of a phenotypic assay monitoring resistance formation to protease inhibitors in HIV-1-infected patients.

    PubMed

    Gehringer, Heike; Von der Helm, Klaus; Seelmeir, Sigrid; Weissbrich, Benedikt; Eberle, Josef; Nitschko, Hans

    2003-05-01

    A novel phenotypic assay, based on recombinant expression of the HIV-1-protease was developed and evaluated; it monitors the formation of resistance to protease inhibitors. The HIV-1 protease-encoding region from the blood sample of patients was amplified, ligated into the expression vector pBD2, and recombinantly expressed in Escherichia coli TG1 cells. The resulting recombinant enzyme was purified by a newly developed one-step acid extraction protocol. The protease activity was determined in presence of five selected HIV protease inhibitors and the 50% inhibitory concentration (IC(50)) to the respective protease inhibitors determined. The degree of resistance was expressed in terms of x-fold increase in IC(50) compared to the IC(50) value of an HIV-1 wild type protease preparation. The established test system showed a reproducible recombinant expression of each individual patients' HIV-1 protease population. Samples of nine clinically well characterised HIV-1-infected patients with varying degrees of resistance were analysed. There was a good correlation between clinical parameters and the results obtained by this phenotypic assay. For the majority of patients a blind genotypic analysis of the patients' protease domain revealed a fair correlation to the results of the phenotypic assay. In a minority of patients our phenotypic results diverged from the genotypic ones. This novel phenotypic assay can be carried out within 8-10 days, and offers a significant advantage in time to the current employed phenotypic tests.

  17. Bioassay battery interlaboratory investigation of emerging contaminants in spiked water extracts - Towards the implementation of bioanalytical monitoring tools in water quality assessment and monitoring.

    PubMed

    Di Paolo, Carolina; Ottermanns, Richard; Keiter, Steffen; Ait-Aissa, Selim; Bluhm, Kerstin; Brack, Werner; Breitholtz, Magnus; Buchinger, Sebastian; Carere, Mario; Chalon, Carole; Cousin, Xavier; Dulio, Valeria; Escher, Beate I; Hamers, Timo; Hilscherová, Klára; Jarque, Sergio; Jonas, Adam; Maillot-Marechal, Emmanuelle; Marneffe, Yves; Nguyen, Mai Thao; Pandard, Pascal; Schifferli, Andrea; Schulze, Tobias; Seidensticker, Sven; Seiler, Thomas-Benjamin; Tang, Janet; van der Oost, Ron; Vermeirssen, Etienne; Zounková, Radka; Zwart, Nick; Hollert, Henner

    2016-11-01

    Bioassays are particularly useful tools to link the chemical and ecological assessments in water quality monitoring. Different methods cover a broad range of toxicity mechanisms in diverse organisms, and account for risks posed by non-target compounds and mixtures. Many tests are already applied in chemical and waste assessments, and stakeholders from the science-police interface have recommended their integration in regulatory water quality monitoring. Still, there is a need to address bioassay suitability to evaluate water samples containing emerging pollutants, which are a current priority in water quality monitoring. The presented interlaboratory study (ILS) verified whether a battery of miniaturized bioassays, conducted in 11 different laboratories following their own protocols, would produce comparable results when applied to evaluate blinded samples consisting of a pristine water extract spiked with four emerging pollutants as single chemicals or mixtures, i.e. triclosan, acridine, 17α-ethinylestradiol (EE2) and 3-nitrobenzanthrone (3-NBA). Assays evaluated effects on aquatic organisms from three different trophic levels (algae, daphnids, zebrafish embryos) and mechanism-specific effects using in vitro estrogenicity (ER-Luc, YES) and mutagenicity (Ames fluctuation) assays. The test battery presented complementary sensitivity and specificity to evaluate the different blinded water extract spikes. Aquatic organisms differed in terms of sensitivity to triclosan (algae > daphnids > fish) and acridine (fish > daphnids > algae) spikes, confirming the complementary role of the three taxa for water quality assessment. Estrogenicity and mutagenicity assays identified with high precision the respective mechanism-specific effects of spikes even when non-specific toxicity occurred in mixture. For estrogenicity, although differences were observed between assays and models, EE2 spike relative induction EC 50 values were comparable to the literature, and E2/EE2

  18. Characterization and Disposition of Legacy Low-Level Waste at the Y-12 National Security Complex - 12133

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tharp, Tim; Donnelly, Jim

    2012-07-01

    The Y-12 National Security Complex (Y-12) is concluding a multi-year program to characterize and dispose of all legacy low-level waste (LLW). The inventory of legacy waste at Y-12 has been reduced from over 3500 containers in Fiscal Year (FY) 2000 to 6 containers at the end of FY2011. In addition, the site recently eliminated the inventory of other low-level waste that is greater than 365 days old (i.e., >365-Day LLW), to be in full compliance with DOE Order 435.1. A consistent technical characterization approach emerged for both of these populations of backlogged waste: (1) compile existing historical data and processmore » knowledge and conduct interviews with site personnel; (2) inspect the containers and any tags, labels, or other markings to confirm or glean additional data; (3) with appropriate monitoring, open the container, visually inspect and photograph the contents while obtaining preliminary radiological surveys; (4) obtain gross weight and field non-destructive assay (NDA) data as needed; (5) use the non-public Oak Ridge Reservation Haul Road to ship the container to a local offsite vendor for waste sorting and segregation; (6) sort, drain, sample, and remove prohibited items; and (7) compile final data and prepare for shipment to disposal. After disposing of this backlog, the focus has now turned to avoiding the recurrence of this situation by maintaining low inventories of low-level waste and shortening the duration between waste generation and disposal. An enhanced waste tracking system and monthly metric charts are used to monitor and report progress to contractor and federal site office management. During the past 2 years, the average age of LLW onsite at Y-12 has decreased from more than 180 days to less than 60 days. (authors)« less

  19. First Industrial Tests of a Matrix Monitor Correction for the Differential Die-away Technique of Historical Waste Drums

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Antoni, Rodolphe; Passard, Christian; Perot, Bertrand

    2015-07-01

    The fissile mass in radioactive waste drums filled with compacted metallic residues (spent fuel hulls and nozzles) produced at AREVA NC La Hague reprocessing plant is measured by neutron interrogation with the Differential Die-away measurement Technique (DDT). In the next years, old hulls and nozzles mixed with Ion-Exchange Resins will be measured. The ion-exchange resins increase neutron moderation in the matrix, compared to the waste measured in the current process. In this context, the Nuclear Measurement Laboratory (LMN) of CEA Cadarache has studied a matrix effect correction method, based on a drum monitor, namely a 3He proportional counter located insidemore » the measurement cavity. After feasibility studies performed with LMN's PROMETHEE 6 laboratory measurement cell and with MCNPX simulations, this paper presents first experimental tests performed on the industrial ACC (hulls and nozzles compaction facility) measurement system. A calculation vs. experiment benchmark has been carried out by performing dedicated calibration measurements with a representative drum and {sup 235}U samples. The comparison between calculation and experiment shows a satisfactory agreement for the drum monitor. The final objective of this work is to confirm the reliability of the modeling approach and the industrial feasibility of the method, which will be implemented on the industrial station for the measurement of historical wastes. (authors)« less

  20. Identification of inhibitors using a cell-based assay for monitoring Golgi-resident protease activity.

    PubMed

    Coppola, Julia M; Hamilton, Christin A; Bhojani, Mahaveer S; Larsen, Martha J; Ross, Brian D; Rehemtulla, Alnawaz

    2007-05-01

    Noninvasive real-time quantification of cellular protease activity allows monitoring of enzymatic activity and identification of activity modulators within the protease's natural milieu. We developed a protease activity assay based on differential localization of a recombinant reporter consisting of a Golgi retention signal and a protease cleavage sequence fused to alkaline phosphatase (AP). When expressed in mammalian cells, this protein localizes to Golgi bodies and, on protease-mediated cleavage, AP translocates to the extracellular medium where its activity is measured. We used this system to monitor the Golgi-associated protease furin, a pluripotent enzyme with a key role in tumorigenesis, viral propagation of avian influenza, ebola, and HIV as well as in activation of anthrax, pseudomonas, and diphtheria toxins. This technology was adapted for high-throughput screening of 39,000-compound small molecule libraries, leading to identification of furin inhibitors. Furthermore, this strategy was used to identify inhibitors of another Golgi protease, the beta-site amyloid precursor protein (APP)-cleaving enzyme (BACE). BACE cleavage of the APP leads to formation of the Abeta peptide, a key event that leads to Alzheimer's disease. In conclusion, we describe a customizable noninvasive technology for real-time assessment of Golgi protease activity used to identify inhibitors of furin and BACE.

  1. Identification of inhibitors using a cell based assay for monitoring golgi-resident protease activity

    PubMed Central

    Coppola, Julia M.; Hamilton, Christin A.; Bhojani, Mahaveer S.; Larsen, Martha J.; Ross, Brian D.; Rehemtulla, Alnawaz

    2007-01-01

    Non-invasive real time quantification of cellular protease activity allows monitoring of enzymatic activity and identification of activity modulators within the protease’s natural milieu. We developed a protease-activity assay based on differential localization of a recombinant reporter consisting of a Golgi retention signal and a protease cleavage sequence fused to alkaline phosphatase (AP). When expressed in mammalian cells, this protein localizes to Golgi bodies and, upon protease mediated cleavage, AP translocates to the extracellular medium where its activity is measured. We used this system to monitor the Golgi-associated protease furin, a pluripotent enzyme with a key role in tumorigenesis, viral propagation of avian influenza, ebola, and HIV, and in activation of anthrax, pseudomonas, and diphtheria toxins. This technology was adapted for high throughput screening of 30,000 compound small molecule libraries, leading to identification of furin inhibitors. Further, this strategy was utilized to identify inhibitors of another Golgi protease, the β-site APP-cleaving enzyme (BACE). BACE cleavage of the amyloid precursor protein leads to formation of the Aβ peptide, a key event that leads to Alzheimer’s disease. In conclusion, we describe a customizable, non-invasive technology for real time assessment of Golgi protease activity used to identify inhibitors of furin and BACE. PMID:17316541

  2. Transboundary hazardous waste management. Part I: Waste management policy of importing countries.

    PubMed

    Fan, Kuo-Shuh; Chang, Tien Chin; Ni, Shih-Piao; Lee, Ching-Hwa

    2005-12-01

    Mixed metal-containing waste, polychlorinated biphenyls (PCB) containing capacitors, printed circuit boards, steel mill dust and metal sludge were among the most common wastes exported from Taiwan. Before the implementation of the self-monitoring model programme of the Basel Convention (secretariat of the Basel Convention 2001) in the Asia region, Taiwan conducted a comprehensive 4-year follow-up project involving government authorities and the waste disposal facilities of the importing countries. A total of five countries and nine plants were visited in 2001-2002. The following outcomes can be drawn from these investigations. The Chinese government adopts the strategies of 'on-site processing' and 'relative centralization' on the waste management by tightening permitting and increasing site inspection. A three-level reviewing system is adopted for the import application. The United States have not signed the Basel Convention yet; the procedures of hazardous waste import rely on bilateral agreements. Importers are not required to provide official notification from the waste exporting countries. The operation, administration, monitoring and licensing of waste treatment plants are governed by the state environmental bureau. Finland, France and Belgium are members of the European Union. The procedures and policies of waste import are similar. All of the documents associated with transboundary movement require the approval of each government involved. Practically, the notification forms and tracking forms effectively manage the waste movement.

  3. Enzyme and inhibition assay of urease by continuous monitoring of the ammonium formation based on capillary electrophoresis.

    PubMed

    Liu, Xiaoxia; Yang, Jiqing; Sun, Shucheng; Guo, Liping; Yang, Li

    2016-10-01

    We present here an easy-to-operate and efficient method for enzyme and inhibition assays of urease, which is a widely distributed and important enzyme that catalyzes the hydrolysis of urea to ammonia and CO 2 . The assay was achieved by integrating CE technique and rapid on-line derivatization method, allowing us to continuously drive the sample to the capillary, thus to measure the amount of the product ammonia from the beginning to the end of the reaction. The method exhibits excellent repeatability with RSD as low as 2.5% for the initial reaction rate (n = 5), with the LOD of ammonia of 20 μM (S/N = 5). The enzyme activity as well as the inhibition of urease by Cu 2+ were investigated using the present method. The results show that Cu 2+ is a noncompetitive inhibitor on urease, in accordance with the result published in the literature. The enzyme activity and inhibition kinetic constants were obtained and were found to be consistent with the results of traditional off-line enzyme assays. Our study indicates that the present approach is a reliable and convenient method for analysis of the urease activity and inhibition kinetics by continuous on-line monitoring of the ammonium formation based on CE. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Current status of therapeutic drug monitoring in Australia and New Zealand: a need for improved assay evaluation, best practice guidelines, and professional development.

    PubMed

    Norris, Ross L; Martin, Jennifer H; Thompson, Erin; Ray, John E; Fullinfaw, Robert O; Joyce, David; Barras, Michael; Jones, Graham R; Morris, Raymond G

    2010-10-01

    The measurement of drug concentrations, for clinical purposes, occurs in many diagnostic laboratories throughout Australia and New Zealand. However, the provision of a comprehensive therapeutic drug monitoring (TDM) service requires the additional elements of pre- and postanalytical advice to ensure that concentrations reported are meaningful, interpretable, and clinically applicable to the individual patient. The aim of this project was to assess the status of TDM services in Australia and New Zealand. A range of professions involved in key aspects of TDM was surveyed by questionnaire in late 2007. Information gathered included: the list of drugs assayed; analytical methods used; interpretation services offered; interpretative methods used; and further monitoring advice provided. Fifty-seven responses were received, of which 42% were from hospitals (public and/or private); 11% a hospital (public and/or private) and pathology provider; and 47% a pathology provider only (public and/or private). Results showed that TDM is applied to a large number of different drugs. Poorly performing assay methods were used in some cases, even when published guidelines recommended alternative practices. Although there was a wide array of assays available, the evidence suggested a need for better selection of assay methods. In addition, only limited advice and/or interpretation of results was offered. Of concern, less than 50% of those providing advice on aminoglycoside dosing in adults used pharmacokinetic tools with six of 37 (16.2%) respondents using Bayesian pharmacokinetic tools, the method recommended in the Australian Therapeutic Guidelines: Antibiotic. In conclusion, the survey highlighted deficiencies in the provision of TDM services, in particular assay method selection and both quality and quantity of postanalytical advice. A range of recommendations, some of which may have international implications, are discussed. There is a need to include measures of impact on clinical

  5. Bioremediation of cooking oil waste using lipases from wastes

    PubMed Central

    do Prado, Débora Zanoni; Facanali, Roselaine; Marques, Márcia Mayo Ortiz; Nascimento, Augusto Santana; Fernandes, Célio Junior da Costa; Zambuzzi, William Fernando

    2017-01-01

    Cooking oil waste leads to well-known environmental impacts and its bioremediation by lipase-based enzymatic activity can minimize the high cytotoxic potential. In addition, they are among the biocatalysts most commercialized worldwide due to the versatility of reactions and substrates. However, although lipases are able to process cooking oil wastes, the products generated from this process do not necessarily become less toxic. Thus, the aim of the current study is to analyze the bioremediation of lipase-catalyzed cooking oil wastes, as well as their effect on the cytotoxicity of both the oil and its waste before and after enzymatic treatment. Thus, assessed the post-frying modification in soybean oil and in its waste, which was caused by hydrolysis reaction catalyzed by commercial and home-made lipases. The presence of lipases in the extracts obtained from orange wastes was identified by zymography. The profile of the fatty acid esters formed after these reactions was detected and quantified through gas chromatography and fatty acids profile compared through multivariate statistical analyses. Finally, the soybean oil and its waste, with and without enzymatic treatment, were assessed for toxicity in cytotoxicity assays conducted in vitro using fibroblast cell culture. The soybean oil wastes treated with core and frit lipases through transesterification reaction were less toxic than the untreated oils, thus confirming that cooking oil wastes can be bioremediated using orange lipases. PMID:29073166

  6. Bioremediation of cooking oil waste using lipases from wastes.

    PubMed

    Okino-Delgado, Clarissa Hamaio; Prado, Débora Zanoni do; Facanali, Roselaine; Marques, Márcia Mayo Ortiz; Nascimento, Augusto Santana; Fernandes, Célio Junior da Costa; Zambuzzi, William Fernando; Fleuri, Luciana Francisco

    2017-01-01

    Cooking oil waste leads to well-known environmental impacts and its bioremediation by lipase-based enzymatic activity can minimize the high cytotoxic potential. In addition, they are among the biocatalysts most commercialized worldwide due to the versatility of reactions and substrates. However, although lipases are able to process cooking oil wastes, the products generated from this process do not necessarily become less toxic. Thus, the aim of the current study is to analyze the bioremediation of lipase-catalyzed cooking oil wastes, as well as their effect on the cytotoxicity of both the oil and its waste before and after enzymatic treatment. Thus, assessed the post-frying modification in soybean oil and in its waste, which was caused by hydrolysis reaction catalyzed by commercial and home-made lipases. The presence of lipases in the extracts obtained from orange wastes was identified by zymography. The profile of the fatty acid esters formed after these reactions was detected and quantified through gas chromatography and fatty acids profile compared through multivariate statistical analyses. Finally, the soybean oil and its waste, with and without enzymatic treatment, were assessed for toxicity in cytotoxicity assays conducted in vitro using fibroblast cell culture. The soybean oil wastes treated with core and frit lipases through transesterification reaction were less toxic than the untreated oils, thus confirming that cooking oil wastes can be bioremediated using orange lipases.

  7. A simple reagent-free spectrophotometric assay for monitoring metronidazole therapy in aquarium water.

    PubMed

    Webb, D Harry; Marrero, Cynthia; Ellis, Helen; Merriwether, Lea; Dove, Alistair D M

    2013-09-01

    A reagent-free spectrophotometric assay was developed to measure the concentration of metronidazole (a 5-nitroimidazole) in both freshwater and seawater matrices. This assay is simple, repeatable, sensitive, and precise and is ideal for use when a rapid, selective test to determine metronidazole concentration in aqueous matrices is necessary. The assay was practically tested on a South American fishes display during treatment with metronidazole for an outbreak of the flagellated parasite Spironucleus in a mixed cichlid (family Cichlidae) and tetra (family Characidae) community. The assay clearly illustrated the course of treatment for the system during a real clinical application. The assay is not without limitations, as interferences can occur from other drugs in the matrix with similar absorbance spectra. Nonetheless, this type of assay illustrates the potential for use of native absorbance assays in aqueous matrices for this and other therapeutic compounds.

  8. A highly rapid and simple competitive enzyme-linked immunosorbent assay for monitoring paralytic shellfish poisoning toxins in shellfish.

    PubMed

    Kawatsu, Kentaro; Kanki, Masashi; Harada, Tetsuya; Kumeda, Yuko

    2014-11-01

    Using a streptavidin-coated well plate, a biotin-labelled anti-gonyautoxin 2/3 monoclonal antibody GT-13A, and a decarbamoyl saxitoxin-peroxidase conjugate, a direct competitive enzyme-linked immunosorbent assay (PSP-ELISA) was developed for monitoring paralytic shellfish poisoning (PSP) toxins in shellfish. This assay is simple to perform and can be completed in approximately 20 min. The PSP-ELISA was compared to the mouse bioassay (MBA) for the detection of PSP toxins in shellfish samples (n=83) collected from the coast of Osaka Prefecture, Japan. When positive and negative results were indicated based on the regulatory limit for PSP toxins (4 mouse unit(MU)/g of shellfish meat), the PSP-ELISA results showed a sensitivity of 100% (25 of 25) and a specificity of 89.7% (52 of 58 samples) compared to the MBA results. These results suggest that the PSP-ELISA could be used as a rapid and simple screening method prior to the MBA. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. F-Area Hazardous Waste Management Facility groundwater monitoring report, Third and fourth quarters 1995: Volume 1

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    NONE

    1996-03-01

    Groundwater at the F-Area Hazardous Waste Management Facility (HWMF) is monitored in compliance with applicable regulations. Monitoring results are compared to the South Carolina Department of Health and Environmental Control (SCDHEC) Groundwater Protection Standard (GWPS). Historically and currently, gross alpha, nitrates, nonvolatile beta, and tritium are among the primary constituents to exceed standards. Numerous other radionuclides and hazardous constituents also exceed the GWPS in the groundwater during the second half of 1995, notably cadmium, lead, radium-226, radium-228, strontium-90, and total alpha-emitting radium. The elevated constituents were found primarily in the water table (aquifer zone IIB{sub 2}), however, several other aquifermore » unit monitoring wells contained elevated levels of constituents. Water-level maps indicate that the groundwater flow rates and directions at the F-Area HWMF have remained relatively constant since the basins ceased to be active in 1988.« less

  10. Solid Waste Management Units And Areas Of Concern Annual Long-Term Monitoring & Maintenance Report For Calendar Year 2016.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dotson, Patrick Wells; Little, Bonnie Colleen

    Long-term controls were maintained at 21 Solid Waste Management Units (SWMUs) and Areas of Concern (AOCs) in accordance with the requirements of the “Long-Term Monitoring and Maintenance Plan for SWMUs and AOCs Granted Corrective Action Complete with Controls” in Attachment M of the Resource Conservation and Recovery Act Facility Operating Permit, which took effect February 26, 2015. Maintenance and controls at these SWMUs and AOCs are described and documented in this report. Conditions requiring maintenance or repair activities were not identified for any of the inspected SWMUs or AOCs. Based upon the inspections performed and site conditions observed, the administrativemore » and physical institutional controls in place at the SWMUs and AOCs are effectively providing continued protection of human health and the environment. This report does not present monitoring and maintenance activities for SWMU 76, the Mixed Waste Landfill; those activities adhere to the approved MWL LTMM Plan, Section 4.8.1 requiring a separate annual report which will be submitted to the NMED by June 30, 2017.« less

  11. Measurement of radionuclides in waste packages

    DOEpatents

    Brodzinski, R.L.; Perkins, R.W.; Rieck, H.G.; Wogman, N.A.

    1984-09-12

    A method is described for non-destructively assaying the radionuclide content of solid waste in a sealed container by analysis of the waste's gamma-ray spectrum and neutron emissions. Some radionuclides are measured by characteristic photopeaks in the gamma-ray spectrum; transuranic nuclides are measured by neutron emission rate; other radionuclides are measured by correlation with those already measured.

  12. Measurement of radionuclides in waste packages

    DOEpatents

    Brodzinski, Ronald L.; Perkins, Richard W.; Rieck, Henry G.; Wogman, Ned A.

    1986-01-01

    A method is described for non-destructively assaying the radionuclide content of solid waste in a sealed container by analysis of the waste's gamma-ray spectrum and neutron emissions. Some radionuclides are measured by characteristic photopeaks in the gamma-ray spectrum; transuranic nuclides are measured by neutron emission rate; other radionuclides are measured by correlation with those already measured.

  13. Developing Tribal Integrated Waste Management Plans

    EPA Pesticide Factsheets

    An IWMP outlines how the tribe will reduce, manage, and dispose of its waste. It identifies existing waste systems, assesses needs, and sets forth the ways to design, implement, and monitor a more effective and sustainable waste management program.

  14. Surface water sampling and analysis plan for environmental monitoring in Waste Area Grouping 6 at Oak Ridge National Laboratory, Oak Ridge, Tennessee. Environmental Restoration Program

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Not Available

    1994-06-01

    This Sampling and Analysis Plan addresses surface water monitoring, sampling, and analysis activities that will be conducted in support of the Environmental Monitoring Plan for Waste Area Grouping (WAG) 6. WAG 6 is a shallow-burial land disposal facility for low-level radioactive waste at the Oak Ridge National Laboratory, a research facility owned by the US Department of Energy and managed by Martin Marietta Energy Systems, Inc. Surface water monitoring will be conducted at nine sites within WAG 6. Activities to be conducted will include the installation, inspection, and maintenance of automatic flow-monitoring and sampling equipment and manual collection of variousmore » water and sediment samples. The samples will be analyzed for various organic, inorganic, and radiological parameters. The information derived from the surface water monitoring, sampling, and analysis will aid in evaluating risk associated with contaminants migrating off-WAG, and will be used in calculations to establish relationships between contaminant concentration (C) and flow (Q). The C-Q relationship will be used in calculating the cumulative risk associated with the off-WAG migration of contaminants.« less

  15. Toxicity assessment of carbon black waste: A by-product from oil refineries.

    PubMed

    Zhen, Xu; Ng, Wei Cheng; Fendy; Tong, Yen Wah; Dai, Yanjun; Neoh, Koon Gee; Wang, Chi-Hwa

    2017-01-05

    In Singapore, approximately 30t/day of carbon-based solid waste are produced from petrochemical processes. This carbon black waste has been shown to possess physical properties that are characteristic of a good adsorbent such as high external surface area. Therefore, there is a growing interest to reutilize and process this carbon black waste into secondary materials such as adsorbents. However, the carbon black waste obtained from petrochemical industries may contain heavy metals that are hazardous to human health and the environment, hence restricting its full potential for re-utilization. Therefore, it is important to examine the possible toxicity effects and toxicity mechanism of carbon black waste on human health. In this study, inductively coupled plasma optical emission spectroscopy (ICP-OES) analysis showed that the heavy metals, vanadium (V), molybdenum (Mo) and nickel (Ni), were present in the carbon black waste in high concentrations. Three human cell lines (HepG2 cells, MRC-5 cells and MDA-MB-231 cells) were used to investigate the toxicity of carbon black waste extract in a variety of in vitro assays. Results from MTS assays indicated that carbon black waste extract decreased the viability of all three cell lines in a dose and time-dependent manner. Observations from confocal microscopy further confirmed this phenomenon. Flow cytometry assay also showed that carbon black waste extract induced apoptosis of human cell lines, and the level of apoptosis increased with increasing waste concentration. Results from reactive oxygen species (ROS) assay indicated that carbon black waste extract induced oxidative stress by increasing intracellular ROS generation in these three human cell lines. Moreover, induction of oxidative damage in these cells was also observed through the alteration of glutathione (GSH) and superoxide dismutase (SOD) activities. Last but not least, by treating the cells with V-spiked solution of concentration equivalent to that found in the

  16. Raman lidar for hydrogen gas concentration monitoring and future radioactive waste management.

    PubMed

    Liméry, Anasthase; Cézard, Nicolas; Fleury, Didier; Goular, Didier; Planchat, Christophe; Bertrand, Johan; Hauchecorne, Alain

    2017-11-27

    A multi-channel Raman lidar has been developed, allowing for the first time simultaneous and high-resolution profiling of hydrogen gas and water vapor. The lidar measures vibrational Raman scattering in the UV (355 nm) domain. It works in a high-bandwidth photon counting regime using fast SiPM detectors and takes into account the spectral overlap between hydrogen and water vapor Raman spectra. Measurement of concentration profiles of H 2 and H 2 O are demonstrated along a 5-meter-long open gas cell with 1-meter resolution at 85 meters. The instrument precision is investigated by numerical simulation to anticipate the potential performance at longer range. This lidar could find applications in the French project Cigéo for monitoring radioactive waste disposal cells.

  17. A serum and platelet-rich plasma serotonin assay using liquid chromatography tandem mass spectrometry for monitoring of neuroendocrine tumor patients.

    PubMed

    Korse, Catharina M; Buning-Kager, Johanna C G M; Linders, Theodora C; Heijboer, Annemieke C; van den Broek, Daan; Tesselaar, Margot E T; van Tellingen, Olaf; van Rossum, Huub H

    2017-06-01

    Serotonin is used for the diagnosis and follow-up of neuroendocrine tumors (NET). We describe the analytical and clinical validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) based serotonin assay for serum and platelet-rich plasma (PRP). An LC-MS/MS based method for serum and PRP serotonin was validated by determination of assay imprecision, carry-over, linearity, interference, recovery, sample stability and a matrix/method comparison of serum and PRP serotonin was made with whole blood serotonin. Furthermore, upper limits of normal were determined and serotonin concentrations of healthy individuals, 14 NET patients without evidence of disease and 51 NET patients with evidence of disease were compared. For serum and PRP fractions, total assay imprecision was <5%. All correlation coefficients were 0.98 and the serum and platelet-rich serotonin upper limit of normal were 5.5nmol/10 9 platelet and 5.1nmol/10 9 platelet, respectively. NET patients with confirmed evidence of disease had significantly higher serum and PRP serotonin levels when compared to NET patients without evidence of disease and healthy volunteers. LC-MS/MS based serum and PRP serotonin assays were developed with suitable analytical characteristics. Furthermore, serum and PRP serotonin was found to be useful for monitoring NET patients. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Recombinant to modified factor VIII and factor IX - chromogenic and one-stage assays issues.

    PubMed

    Kitchen, S; Kershaw, G; Tiefenbacher, S

    2016-07-01

    The recent development of modified recombinant factor VIII (FVIII) and factor IX (FIX) therapeutic products with extended half-lives will create challenges for the haemostasis laboratory in obtaining recovery estimates of these products in clinical samples using existing assays. The new long-acting therapeutic concentrates contain molecular modifications of Fc fusion, site-specific of polyethylene glycol or albumin fusion. The optimum methods for monitoring each new product will need to be assessed individually and laboratories should select an assay which gives similar results to the assay used to assign potency to the product in question. For some extended half-life FVIII and FIX products some one stage assays are entirely unsuitable for monitoring purposes. For most products and assay reagents studied so far, and reviewed in this manuscript, chromogenic FVIII or FIX assays can be safely used with conventional plasma standards. If one stage assays are used then they should be performed using carefully selected reagents/methods which have been shown to recover activity close to the labelled potency for the specific product being monitored. © 2016 John Wiley & Sons Ltd.

  19. GeneXpert HIV-1 quant assay, a new tool for scale up of viral load monitoring in the success of ART programme in India.

    PubMed

    Kulkarni, Smita; Jadhav, Sushama; Khopkar, Priyanka; Sane, Suvarna; Londhe, Rajkumar; Chimanpure, Vaishali; Dhilpe, Veronica; Ghate, Manisha; Yelagate, Rajendra; Panchal, Narayan; Rahane, Girish; Kadam, Dilip; Gaikwad, Nitin; Rewari, Bharat; Gangakhedkar, Raman

    2017-07-21

    Recent WHO guidelines identify virologic monitoring for diagnosing and confirming ART failure. In view of this, validation and scale up of point of care viral load technologies is essential in resource limited settings. A systematic validation of the GeneXpert® HIV-1 Quant assay (a point-of-care technology) in view of scaling up HIV-1 viral load in India to monitor the success of national ART programme was carried out. Two hundred nineteen plasma specimens falling in nine viral load ranges (<40 to >5 L copies/ml) were tested by the Abbott m2000rt Real Time and GeneXpert HIV-1 Quant assays. Additionally, 20 seronegative; 16 stored specimens and 10 spiked controls were also tested. Statistical analysis was done using Stata/IC and sensitivity, specificity, PPV, NPV and %misclassification rates were calculated as per DHSs/AISs, WHO, NACO cut-offs for virological failure. The GeneXpert assay compared well with the Abbott assay with a higher sensitivity (97%), specificity (97-100%) and concordance (91.32%). The correlation between two assays (r = 0.886) was statistically significant (p < 0.01), the linear regression showed a moderate fit (R 2  = 0.784) and differences were within limits of agreement. Reproducibility showed an average variation of 4.15 and 3.52% while Lower limit of detection (LLD) and Upper limit of detection (ULD) were 42 and 1,740,000 copies/ml respectively. The misclassification rates for three viral load cut offs were not statistically different (p = 0.736). All seronegative samples were negative and viral loads of the stored samples showed a good fit (R 2  = 0.896 to 0.982). The viral load results of GeneXpert HIV-1 Quant assay compared well with Abbott HIV-1 m2000 Real Time PCR; suggesting its use as a Point of care assay for viral load estimation in resource limited settings. Its ease of performance and rapidity will aid in timely diagnosis of ART failures, integrated HIV-TB management and will facilitate the UNAIDS 90-90-90 target.

  20. Polyhydroxybutyrate (PHB) Synthesis by Spirulina sp. LEB 18 Using Biopolymer Extraction Waste.

    PubMed

    da Silva, Cleber Klasener; Costa, Jorge Alberto Vieira; de Morais, Michele Greque

    2018-01-20

    The reuse of waste as well as the production of biodegradable compounds has for years been the object of studies and of global interest as a way to reduce the environmental impact generated by unsustainable exploratory processes. The conversion of linear processes into cyclical processes has environmental and economic advantages, reducing waste deposition and reducing costs. The objective of this work was to use biopolymer extraction waste in the cultivation of Spirulina sp. LEB 18, for the cyclic process of polyhydroxybutyrate (PHB) synthesis. Concentrations of 10, 15, 20, 25, and 30% (v/v) of biopolymer extraction waste were tested. For comparison, two assays were used without addition of waste, Zarrouk (SZ) and modified Zarrouk (ZM), with reduction of nitrogen. The assays were carried out in triplicate and evaluated for the production of microalgal biomass and PHB. The tests with addition of waste presented a biomass production statistically equal to ZM (0.79 g L -1 ) (p < 0.1). The production of PHB in the assay containing 25% of waste was higher when compared to the other cultivations, obtaining 10.6% (w/w) of biopolymer. From the results obtained, it is affirmed that the use of PHB extraction waste in the microalgal cultivation, aiming at the synthesis of biopolymers, can occur in a cyclic process, reducing process costs and the deposition of waste, thus favoring the preservation of the environment.

  1. Basic design of MRM assays for peptide quantification.

    PubMed

    James, Andrew; Jorgensen, Claus

    2010-01-01

    With the recent availability and accessibility of mass spectrometry for basic and clinical research, the requirement for stable, sensitive, and reproducible assays to specifically detect proteins of interest has increased. Multiple reaction monitoring (MRM) or selective reaction monitoring (SRM) is a highly selective, sensitive, and robust assay to monitor the presence and amount of biomolecules. Until recently, MRM was typically used for the detection of drugs and other biomolecules from body fluids. With increased focus on biomarkers and systems biology approaches, researchers in the proteomics field have taken advantage of this approach. In this chapter, we will introduce the reader to the basic principle of designing and optimizing an MRM workflow. We provide examples of MRM workflows for standard proteomic samples and provide suggestions for the reader who is interested in using MRM for quantification.

  2. Advances in Developing HIV-1 Viral Load Assays for Resource-Limited Settings

    PubMed Central

    Wang, ShuQi; Xu, Feng; Demirci, Utkan

    2010-01-01

    Commercial HIV-1 RNA viral load assays have been routinely used in developed countries to monitor antiretroviral treatment (ART). However, these assays require expensive equipment and reagents, well-trained operators, and established laboratory infrastructure. These requirements restrict their use in resource-limited settings where people are most afflicted with the HIV-1 epidemic. Inexpensive alternatives such as the Ultrasensitive p24 assay, the Reverse Transcriptase (RT) assay and in-house reverse transcription quantitative polymerase chain reaction (RT-qPCR) have been developed. However, they are still time-consuming, technologically complex and inappropriate for decentralized laboratories as point-of-care (POC) tests. Recent advances in microfluidics and nanotechnology offer new strategies to develop low-cost, rapid, robust and simple HIV-1 viral load monitoring systems. We review state-of-the-art technologies used for HIV-1 viral load monitoring in both developed and developing settings. Emerging approaches based on microfluidics and nanotechnology, which have potential to be integrated into POC HIV-1 viral load assays, are also discussed. PMID:20600784

  3. 40 CFR 258.51 - Ground-water monitoring systems.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 26 2013-07-01 2013-07-01 false Ground-water monitoring systems. 258.51 Section 258.51 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS Ground-Water Monitoring and Corrective Action § 258.51 Ground-water monitoring systems. (a) A...

  4. Lessons Learned from Radioactive Waste Storage and Disposal Facilities

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Esh, David W.; Bradford, Anna H.

    2008-01-15

    The safety of radioactive waste disposal facilities and the decommissioning of complex sites may be predicated on the performance of engineered and natural barriers. For assessing the safety of a waste disposal facility or a decommissioned site, a performance assessment or similar analysis is often completed. The analysis is typically based on a site conceptual model that is developed from site characterization information, observations, and, in many cases, expert judgment. Because waste disposal facilities are sited, constructed, monitored, and maintained, a fair amount of data has been generated at a variety of sites in a variety of natural systems. Thismore » paper provides select examples of lessons learned from the observations developed from the monitoring of various radioactive waste facilities (storage and disposal), and discusses the implications for modeling of future waste disposal facilities that are yet to be constructed or for the development of dose assessments for the release of decommissioning sites. Monitoring has been and continues to be performed at a variety of different facilities for the disposal of radioactive waste. These include facilities for the disposal of commercial low-level waste (LLW), reprocessing wastes, and uranium mill tailings. Many of the lessons learned and problems encountered provide a unique opportunity to improve future designs of waste disposal facilities, to improve dose modeling for decommissioning sites, and to be proactive in identifying future problems. Typically, an initial conceptual model was developed and the siting and design of the disposal facility was based on the conceptual model. After facility construction and operation, monitoring data was collected and evaluated. In many cases the monitoring data did not comport with the original site conceptual model, leading to additional investigation and changes to the site conceptual model and modifications to the design of the facility. The following cases are

  5. Five-year performance monitoring of a high-density polyethylene (HDPE) cover system at a reclaimed mine waste rock pile in the Sydney Coalfield (Nova Scotia, Canada).

    PubMed

    Power, Christopher; Ramasamy, Murugan; MacAskill, Devin; Shea, Joseph; MacPhee, Joseph; Mayich, David; Baechler, Fred; Mkandawire, Martin

    2017-12-01

    Cover systems are commonly placed over waste rock piles (WRPs) to limit atmospheric water and oxygen ingress and control the generation and release of acid mine drainage (AMD) to the receiving environment. Although covers containing geomembranes such as high-density polyethylene (HDPE) exhibit the attributes to be highly effective, there are few, if any, published studies monitoring their performance at full-scale WRPs. In 2011, a HDPE cover was installed over the Scotchtown Summit WRP in Nova Scotia, Canada, and extensive field performance monitoring was conducted over the next five years. A range of parameters within the atmosphere, cover, waste rock, groundwater and surface water, were monitored and integrated into a comprehensive hydrogeochemical conceptual model to assess (i) atmospheric ingress to the waste rock, (ii) waste rock acidity and depletion and (iii) evolution of groundwater and surface water quality. Results demonstrate that the cover is effective and meeting site closure objectives. Depletion in oxygen influx resulted in slower sulphide oxidation and AMD generation, while a significant reduction in water influx (i.e. 512 to 50 mm/year) resulted in diminished AMD release. Consistent improvements in groundwater quality (decrease in sulphate and metals; increase in pH) beneath and downgradient of the WRP were observed. Protection and/or significant improvement in surface water quality was evident in all surrounding watercourses due to the improved groundwater plume and elimination of contaminated runoff over previously exposed waste rock. A variably saturated flow and contaminant transport model is currently being developed to predict long-term cover system performance.

  6. Using RT-PCR and bDNA assays to measure non-clade B HIV-1 subtype RNA.

    PubMed

    Pasquier, C; Sandres, K; Salama, G; Puel, J; Izopet, J

    1999-08-01

    The performance of the new version of RT-PCR assay (Amplicor HIV-1 Monitor v1.5) was assessed. The quantification of non-B subtype HIV-1 plasma RNA (30A, 1C, 1D, 3E, 2F, 3G) obtained using Monitor v1.5 was compared to the former version of this assay (Monitor v1.0) and to the Quantiplex v2.0 bDNA assay. The new primers used in Monitor v1.5 were similar to the former version in both specificity and sensitivity. The new primers corrected the detection and quantification defect observed previously for HIV-1 non-B subtypes and gave slightly higher RNA concentrations than those measured using the bDNA assay (+0.39 log copies/ml).

  7. Multicenter evaluation of the Bayer Immuno I CA 15-3 assay.

    PubMed

    Cheli, C D; Morris, D L; Kish, L; Goldblatt, J; Neaman, I; Allard, W J; Yeung, K K; Wu, A H; Moore, R; Chan, D W; Fritsche, H A; Schwartz, M K; Very, D L

    1998-04-01

    We conducted a multicenter evaluation of the analytical and clinical features of the automated Bayer Immuno 1 CA 15-3 assay and compared assay performance to two manual tests. Results of the 10-day imprecision study of the Bayer Immuno 1 assay pooled across four evaluation sites and three lots of reagent produced total CV < or = 4%. Lot-to-lot reproducibility for 26 different lots of reagents and calibrators manufactured over a 2-year period was demonstrated (CV, 1.1%). Results for the Bayer Immuno 1 assay correlated well with the Biomira TRUQUANT BR 27.29 and Centocor CA 15-3 RIAs (r > or = 0.94). The upper limit of the reference interval for the Bayer Immuno 1 assay was 35.9 kilounits/L (35.9 units/mL); values were similar for all methods. Longitudinal monitoring of healthy women yielded assay values with an average CV of 11% and 21% for the Bayer Immuno 1 and Biomira assays, respectively. The Bayer Immuno 1 assay demonstrated the analytical features, intermethod correlation, and long-term performance characteristics that are essential for longitudinal monitoring of breast cancer patients.

  8. Biomass conversion determined via fluorescent cellulose decay assay.

    PubMed

    Wischmann, Bente; Toft, Marianne; Malten, Marco; McFarland, K C

    2012-01-01

    An example of a rapid microtiter plate assay (fluorescence cellulose decay, FCD) that determines the conversion of cellulose in a washed biomass substrate is reported. The conversion, as verified by HPLC, is shown to correlate to the monitored FCD in the assay. The FCD assay activity correlates to the performance of multicomponent enzyme mixtures and is thus useful for the biomass industry. The development of an optimized setup of the 96-well microtiter plate is described, and is used to test a model that shortens the assay incubation time from 72 to 24h. A step-by-step procedure of the final assay is described. Copyright © 2012 Elsevier Inc. All rights reserved.

  9. Cytogenetic status of healthy children assessed with the alkaline comet assay and the cytokinesis-block micronucleus cytome assay.

    PubMed

    Gajski, Goran; Gerić, Marko; Oreščanin, Višnja; Garaj-Vrhovac, Vera

    2013-01-20

    In the present study the alkaline comet assay and the cytokinesis-block micronucleus cytome (CBMN Cyt) assay were used to evaluate the baseline frequency of cytogenetic damage in peripheral blood lymphocytes (PBLs) of 50 healthy children from the general population in Croatia (age, 11.62±1.81 years). Mean values of tail length, tail intensity and tail moment, as comet assay parameters, were 12.92±0.10, 0.73±0.06 and 0.08±0.01, respectively. The mean frequency of micronuclei (MN) for all subjects was 2.32±0.28 per 1000 bi-nucleated cells, while the mean frequency of nucleoplasmic bridges (NPBs) was 1.72±0.24 and of nuclear buds (NBUDs) 1.44±0.19. The mean nuclear division index (NDI) was 1.70±0.05. When comet-assay parameters were considered, higher mean values for all three were found for the female population. According to the Mann-Whitney U test applied on the results of the comet assay, the only statistically significant difference between the male and female populations was found for tail length. Similar to the results obtained by the comet assay, girls showed higher mean values of all three measured parameters of the CBMN Cyt assay. This difference was statistically significant for total number of NPBs only. In the case of the NDI, a higher mean value was also obtained in girls, but this difference was not statistically significant. The results obtained present background data that could be considered as normal values for healthy children living in urban areas, and can later on serve as baseline values for further toxicological monitoring. Additionally, the usefulness of both techniques in measuring cytogenetic damage during bio-monitoring of children is confirmed. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Physicochemical and immunochemical assays for monitoring consistent production of tetanus toxoid.

    PubMed

    Metz, Bernard; Tilstra, Wichard; van der Put, Robert; Spruit, Nanda; van den Ijssel, Jan; Robert, Jolanda; Hendriksen, Coenraad; Kersten, Gideon

    2013-07-01

    The detoxification of tetanus toxin by formaldehyde is a crucial step in the production of tetanus toxoid. The inactivation results in chemically modified proteins and it determines largely the ultimate efficacy and safety of the vaccine. Currently, the quality of tetanus toxoid lots is evaluated in potency and safety tests performed in animals. As a possible alternative, this article describes a panel of in vitro methods, which provides detailed information about the quality of tetanus toxoid. Ten experimental lots of tetanus toxoid were prepared using increasing concentrations of formaldehyde and glycine to obtain tetanus toxoids having differences in antigenicity, immunogenicity, residual toxicity and protein structure. The structural properties of each individual toxoid were determined using immunochemical and physicochemical methods, including biosensor analysis, ELISA, circular dichroism, TNBS assay, differential scanning calorimetry, fluorescence and SDS-PAGE. The quality of a tetanus toxoid lot can be assessed by these set of analytical techniques. Based on antigenicity, immunogenicity and residual toxicity data, criteria are formulated that tetanus toxoids lot have to meet in order to have a high quality. The in vitro methods are a valuable selection of techniques for monitoring consistency of production of tetanus toxoid, especially for the detoxification process of tetanus toxin. Copyright © 2013 The International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  11. M-Area and Metallurgical Laboratory Hazardous Waste Management Facilities groundwater monitoring and corrective-action report (U). Third and fourth quarters 1996, Vol. I

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    NONE

    1997-03-01

    This report describes the groundwater monitoring and corrective-action program at the M-Area Hazardous Waste Management Facility (HWMF) and the Metallurgical Laboratory (Met Lab) HWMF at the Savannah River Site (SRS) during 1996.

  12. FIREFLY LUCIFERASE ATP ASSAY DEVELOPMENT FOR MONITORING BACTERIAL CONCENTRATIONS IN WATER SUPPLIES

    EPA Science Inventory

    This research program was initiated to develop a rapid, automatable system for measuring total viable microorganisms in potable drinking water supplies using the firefly luciferase ATP assay. The assay was adapted to an automatable flow system that provided comparable sensitivity...

  13. Development of Automated Monitoring and Management System of Municipal Solid Waste Landfill Based on the Industrial OMRON Controller

    NASA Astrophysics Data System (ADS)

    Kostarev, S. N.; Sereda, T. G.

    2018-01-01

    The application of the programmable logic integrated circuits (PLC) for creating the software and hardware complexes of the medium complexity is an economically sound solution. The application of the OMRON controller to solve the monitoring and management tasks of safety of the municipal solid waste (MSW) landfill with the use of technology of the filtrate recirculation and the landfill maps irrigation is shown in the article. The article contains the technical solution connected with the implementation of the 2162059RU invention patent for the municipal solid waste landfill management in the Kurgan region of Russia. The calculation of maps and ponds was made with consideration of the limited sanitary and protection zone. The GRUNDFOS dosing and reactor equipment was proposed to use in the project.

  14. Quantitative Analysis of Energy Metabolic Pathways in MCF-7 Breast Cancer Cells by Selected Reaction Monitoring Assay*

    PubMed Central

    Drabovich, Andrei P.; Pavlou, Maria P.; Dimitromanolakis, Apostolos; Diamandis, Eleftherios P.

    2012-01-01

    To investigate the quantitative response of energy metabolic pathways in human MCF-7 breast cancer cells to hypoxia, glucose deprivation, and estradiol stimulation, we developed a targeted proteomics assay for accurate quantification of protein expression in glycolysis/gluconeogenesis, TCA cycle, and pentose phosphate pathways. Cell growth conditions were selected to roughly mimic the exposure of cells in the cancer tissue to the intermittent hypoxia, glucose deprivation, and hormonal stimulation. Targeted proteomics assay allowed for reproducible quantification of 76 proteins in four different growth conditions after 24 and 48 h of perturbation. Differential expression of a number of control and metabolic pathway proteins in response to the change of growth conditions was found. Elevated expression of the majority of glycolytic enzymes was observed in hypoxia. Cancer cells, as opposed to near-normal MCF-10A cells, exhibited significantly increased expression of key energy metabolic pathway enzymes (FBP1, IDH2, and G6PD) that are known to redirect cellular metabolism and increase carbon flux through the pentose phosphate pathway. Our quantitative proteomic protocol is based on a mass spectrometry-compatible acid-labile detergent and is described in detail. Optimized parameters of a multiplex selected reaction monitoring (SRM) assay for 76 proteins, 134 proteotypic peptides, and 401 transitions are included and can be downloaded and used with any SRM-compatible mass spectrometer. The presented workflow is an integrated tool for hypothesis-driven studies of mammalian cells as well as functional studies of proteins, and can greatly complement experimental methods in systems biology, metabolic engineering, and metabolic transformation of cancer cells. PMID:22535206

  15. Mass spectrometric real-time monitoring of an enzymatic phosphorylation assay using internal standards and data-handling freeware.

    PubMed

    Krappmann, Michael; de Boer, Arjen R; Kool, Daniël R W; Irth, Hubertus; Letzel, Thomas

    2016-04-30

    Continuous-flow reaction detection systems (monitoring enzymatic reactions with mass spectrometry (MS)) lack quantitative values so far. Therefore, two independent internal standards (IS) are implemented in a way that the online system stability can be observed, quantitative conversion values for substrate and product can be obtained and they can be used as mass calibration standards for high MS accuracy. An application previously developed for the MS detection of peptide phosphorylation by cAMP-dependent protein kinase A (PKA) (De Boer et al., Anal. Bioanal. Chem. 2005, 381, 647-655) was transferred to a continuous-flow reaction detection system. This enzymatic reaction, involving enzyme activation as well as the transfer of a phosphate group from ATP to a peptide substrate, was used to prove the compatibility of a quantitative enzymatic assay in a continuous-flow real-time system (connected to MS). Moreover (using internal standards), the critical parameter reaction temperature (including solution density variations depending on temperature) was studied in the continuous-flow mixing system. Furthermore, two substrates (malantide and kemptide), two enzyme types (catalytic subunit of PKA and complete PKA) and one inhibitor were tested to determine system robustness and long-term availability. Even spraying solutions that contained significant amount of MS contaminants (e.g. the polluted catalytic subunit) resulted in quantifiable MS signal intensities. Subsequent recalculations using the internal standards led to results representing the power of this application. The presented methodology and the data evaluation with available Achroma freeware enable the direct coupling of biochemical assays with quantitative MS detection. Monitoring changes such as temperature, reaction time, inhibition, or compound concentrations can be observed quantitatively and thus enzymatic activity can be calculated. Copyright © 2016 John Wiley & Sons, Ltd.

  16. Proactive therapeutic drug monitoring of infliximab: a comparative study of a new point-of-care quantitative test with two established ELISA assays.

    PubMed

    Afonso, J; Lopes, S; Gonçalves, R; Caldeira, P; Lago, P; Tavares de Sousa, H; Ramos, J; Gonçalves, A R; Ministro, P; Rosa, I; Vieira, A I; Dias, C C; Magro, F

    2016-10-01

    Therapeutic drug monitoring is a powerful strategy known to improve the clinical outcomes and to optimise the healthcare resources in the treatment of autoimmune diseases. Currently, most of the methods commercially available for the quantification of infliximab (IFX) are ELISA-based, with a turnaround time of approximately 8 h, and delaying the target dosage adjustment to the following infusion. To validate the first point-of-care IFX quantification device available in the market - the Quantum Blue Infliximab assay (Buhlmann, Schonenbuch, Switzerland) - by comparing it with two well-established methods. The three methods were used to assay the IFX concentration of spiked samples and of the serum of 299 inflammatory bowel diseases (IBD) patients undergoing IFX therapy. The point-of-care assay had an average IFX recovery of 92%, being the most precise among the tested methods. The Intraclass Correlation Coefficients of the point-of-care IFX assay vs. the two ELISA-based established methods were 0.889 and 0.939. Moreover, the accuracy of the point-of-care IFX compared with each of the two reference methods was 77% and 83%, and the kappa statistics revealed a substantial agreement (0.648 and 0.738). The Quantum Blue IFX assay can successfully replace the commonly used ELISA-based IFX quantification kits. This point-of-care IFX assay is able to deliver the results within 15 min makes it ideal for an immediate target concentration adjusted dosing. Moreover, it is a user-friendly desktop device that does not require specific laboratory facilities or highly specialised personnel. © 2016 John Wiley & Sons Ltd.

  17. Factors influencing naproxen metabolite interference in total bilirubin assays.

    PubMed

    Saifee, Nabiha Huq; Ranjitkar, Pratistha; Greene, Dina N

    2016-04-01

    The factors influencing naproxen metabolite O-desmethylnaproxen (ODMN) positive interference in diazo-based Jendrassik and Grof (JG) total bilirubin (Tbil) assays and lack of interference in direct bilirubin (Dbil) assays have not been resolved. The objective of this study was to understand the conditions causing this interference pattern. Pooled normal and ultra-filtered plasma samples spiked with ODMN and naproxen were measured on the Beckman Coulter DxC and AU instruments. Absorbance spectra were obtained for ODMN mixed with Dbil reagent at original and adjusted pH. Absorbance spectra were also obtained for ODMN and bilirubin samples mixed with Tbil assay reagents. ODMN produces a positive interference in the DxC JG Tbil assays, but not the AU Tbil or Dbil assays or the DxC Dbil assay. Neutralizing the acidic pH of AU and DxC Dbil reagents allows ODMN to react with diazo salts. ODMN samples mixed with DxC and AU Tbil reagents produce broad peaks from 450 to 560nm and 400 to 540nm, respectively. The DxC JG Tbil assay monitors a change in absorbance at 520nm close to peak absorbance wavelength of diazo-reacted ODMN, whereas the AU Tbil assay monitors a change in absorbance at 570/660nm, beyond the peak absorbance wavelengths of diazo-reacted ODMN. The acidic pH of diazo-based Dbil assay reagents inhibits the reaction of ODMN with diazo salts. The AU JG Tbil assay is a reliable method to measure Tbil in the setting of naproxen overdose. Copyright © 2015 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  18. Air quality assessment for land disposal of industrial wastes

    NASA Astrophysics Data System (ADS)

    Shen, Thomas T.

    1982-07-01

    Air pollution from hazardous waste landfills and lagoons is largely unknown. Routine monitoring of toxic air contaminants associated with hazardous waste facilities is difficult and very costly. The method presented in this paper would be useful for air quality assessment in the absence of monitoring data. It may be used as a screening process to examine the question of whether or not volatilization is considered to be significant for a given contaminant and also to evaluate permit applications for new hazardous waste facilities concerning waste volatilization problems.

  19. NCI Launches Proteomics Assay Portal | Office of Cancer Clinical Proteomics Research

    Cancer.gov

    In a paper recently published by the journal Nature Methods, Investigators from the National Cancer Institute’s Clinical Proteomic Tumor Analysis Consortium (NCI-CPTAC) announced the launch of a proteomics Assay Portal for multiple reaction monitoring-mass spectrometry (MRM-MS) assays.  This community web-based repository for well-characterized quantitative proteomic assays currently consists of 456 unique peptide assays to 282 unique proteins and ser

  20. Apparatus and method for quantitative assay of generic transuranic wastes from nuclear reactors

    DOEpatents

    Caldwell, J.T.; Kunz, W.E.; Atencio, J.D.

    1982-03-31

    A combination of passive and active neutron measurements which yields quantitative information about the isotopic composition of transuranic wastes from nuclear power or weapons material manufacture reactors is described. From the measurement of prompt and delayed neutron emission and the incidence of two coincidentally emitted neutrons from induced fission of fissile material in the sample, one can quantify /sup 233/U, /sup 235/U and /sup 239/Pu isotopes in waste samples. Passive coincidence counting, including neutron multiplicity measurement and determination of the overall passive neutron flux additionally enables the separate quantitative evaluation of spontaneous fission isotopes such as /sup 240/Pu, /sup 244/Cm and /sup 252/Cf, and the spontaneous alpha particle emitter /sup 241/Am. These seven isotopes are the most important constituents of wastes from nuclear power reactors and once the mass of each isotope present is determined by the apparatus and method of the instant invention, the overall alpha particle activity can be determined to better than 1 nCi/g from known radioactivity data. Therefore, in addition to the quantitative analysis of the waste sample useful for later reclamation purposes, the alpha particle activity can be determined to decide whether permanent low-level burial is appropriate for the waste sample.

  1. Apparatus and method for quantitative assay of generic transuranic wastes from nuclear reactors

    DOEpatents

    Caldwell, John T.; Kunz, Walter E.; Atencio, James D.

    1984-01-01

    A combination of passive and active neutron measurements which yields quantitative information about the isotopic composition of transuranic wastes from nuclear power or weapons material manufacture reactors is described. From the measurement of prompt and delayed neutron emission and the incidence of two coincidentally emitted neutrons from induced fission of fissile material in the sample, one can quantify .sup.233 U, .sup.235 U and .sup.239 Pu isotopes in waste samples. Passive coincidence counting, including neutron multiplicity measurement and determination of the overall passive neutron flux additionally enables the separate quantitative evaluation of spontaneous fission isotopes such as .sup.240 Pu, .sup.244 Cm and .sup.252 Cf, and the spontaneous alpha particle emitter .sup.241 Am. These seven isotopes are the most important constituents of wastes from nuclear power reactors and once the mass of each isotope present is determined by the apparatus and method of the instant invention, the overall alpha particle activity can be determined to better than 1 nCi/g from known radioactivity data. Therefore, in addition to the quantitative analysis of the waste sample useful for later reclamation purposes, the alpha particle activity can be determined to decide whether "permanent" low-level burial is appropriate for the waste sample.

  2. Monitoring of DNA breakage in embryonic stages of the African catfish Clarias gariepinus (Burchell, 1822) after exposure to lead nitrate using alkaline comet assay.

    PubMed

    Osman, Alaa G M; Mekkawy, Imam A; Verreth, Johan; Wuertz, Sven; Kloas, Werner; Kirschbaum, Frank

    2008-12-01

    Increasing lead contamination in Egyptian ecosystems and high lead concentrations in food items have raised concern for human health and stimulated studies on monitoring ecotoxicological impact of lead-caused genotoxicity. In this work, the alkaline comet assay was modified for monitoring DNA strand breakage in sensitive early life stages of the African catfish Clarias gariepinus. Following exposure to 100, 300, and 500 microg/L lead nitrate, DNA strand breakage was quantified in embryos at 30, 48, 96, 144, and 168 h post-fertilization (PFS). For quantitative analysis, four commonly used parameters (tail % DNA, %TDNA; head % DNA, %HDNA; tail length, TL; tail moment, TM) were analyzed in 96 nuclei (in triplicates) at each sampling point. The parameter %TDNA revealed highest resolution and lowest variation. A strong correlation between lead concentration, time of exposure, and DNA strand breakage was observed. Here, genotoxicity detected by comet assay preceded the manifested malformations assessed with conventional histology. Qualitative evaluation was carried out using five categories are as follows: undamaged (%TDNA < or = 10%), low damaged (10% < %TDNA < or = 25%), median damaged (25 < %TDNA < or = 50%), highly damaged (50 < %TDNA < or = 75%), and extremely damaged (%TDNA > 75%) nuclei confirming a dose and time-dependent shift towards increased frequencies of highly and extremely damaged nuclei. A protective capacity provided by a hardened chorion is a an interesting finding in this study as DNA damage in the prehatching stages 30 h-PFS and 48 h-PFS was low in all treatments (qualitative and quantitative analyses). These results clearly show that the comet assay is a sensitive tool for the detection of genotoxicity in vulnerable early life stages of the African catfish and is a method more sensitive than histological parameters for monitoring genotoxic effects. 2008 Wiley Periodicals, Inc.

  3. Multiple reaction monitoring and multiple reaction monitoring cubed based assays for the quantitation of apolipoprotein F.

    PubMed

    Kumar, Abhinav; Gangadharan, Bevin; Zitzmann, Nicole

    2016-10-15

    Apolipoprotein F (APO-F) is a novel low abundance liver fibrosis biomarker and its concentration decreases in human serum and plasma across liver fibrosis stages. Current antibody based assays for APO-F suffer from limitations such as unspecific binding, antibody availability and undetectable target if the protein is degraded; and so an antibody-free assay has the potential to be a valuable diagnostic tool. We report an antibody-free, rapid, sensitive, selective and robust LC-MS/MS (MRM and MRM(3)) method for the detection and quantitation of APO-F in healthy human plasma. With further analysis of clinical samples, this LC-MS based method could be established as the first ever antibody-free biomarker assay for liver fibrosis. We explain the use of Skyline software for peptide selection and the creation of a reference library to aid in true peak identification of endogenous APO-F peptides in digests of human plasma without protein or peptide enrichment. Detection of a glycopeptide using MRM-EPI mode and reduction of interferences using MRM3 are explained. The amount of APO-F in human plasma from a healthy volunteer was determined to be 445.2ng/mL, the coefficient of variation (CV) of precision for 20 injections was <12% and the percentage error of each point along the calibration curve was calculated to be <8%, which is in line with the assay requirements for clinical samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Using ATP-driven bioluminescence assay to monitor microbial safety in a contemporary human cadaver laboratory.

    PubMed

    Benninger, Brion; Maier, Thomas

    2015-03-01

    The objective of this study was to utilize a cost-effective method for assessing the levels of bacterial, yeast, and mold activity during a human dissection laboratory course. Nowadays, compliance with safety regulations is policed by institutions at higher standards than ever before. Fear of acquiring an unknown infection is one of the top concerns of professional healthcare students, and it provokes anti-laboratory anxiety. Human cadavers are not routinely tested for bacteria and viruses prior to embalming. Human anatomy dissecting rooms that house embalmed cadavers are normally cleaned after the dissected cadavers have been removed. There is no evidence that investigators have ever assessed bacterial and fungal activities using adenosine triphosphate (ATP)-driven bioluminescence assays. A literature search was conducted on texts, journals, and websites regarding bacterial, yeast, and mold activities in an active cadaver laboratory. Midway into a clinical anatomy course, ATP bioluminescence assays were used to swab various sites within the dissection room, including entrance and exiting door handles, water taps, cadaver tables, counter tops, imaging material, X-ray box switches, and the cadaver surfaces. The results demonstrated very low activities on cadaver tables, washing up areas, and exiting door handles. There was low activity on counter tops and X-ray boxes. There was medium activity on the entrance door handles. These findings suggest an inexpensive and accurate method for monitoring safety compliance and microbial activity. Students can feel confident and safe in the environment in which they work. © 2014 Wiley Periodicals, Inc.

  5. Development of a versatile lab-on-a-chip enzyme assay platform for pathogen detection in CBRNE scenarios

    NASA Astrophysics Data System (ADS)

    Klemm, Richard; Schattschneider, Sebastian; Jahn, Tobias; Hlawatsch, Nadine; Julich, Sandra; Becker, Holger; Gärtner, Claudia

    2013-05-01

    The ability to integrate complete assays on a microfluidic chip helps to greatly simplify instrument requirements and allows the use of lab-on-a-chip technology in the field. A core application for such field-portable systems is the detection of pathogens in a CBRNE scenario such as permanent monitoring of airborne pathogens, e.g. in metro stations or hospitals etc. As one assay methodology for the pathogen identification, enzymatic assays were chosen. In order evaluate different detection strategies, the realized on-chip enzyme assay module has been designed as a general platform chip. In all application cases, the assays are based on immobilized probes located in microfluidic channels. Therefore a microfluidic chip was realized containing a set of three individually addressable channels, not only for detection of the sample itself also to have a set of references for a quantitative analysis. It furthermore includes two turning valves and a waste container for clear and sealed storage of potential pathogenic liquids to avoid contamination of the environment. All liquids remain in the chip and can be disposed of in proper way subsequently to the analysis. The chip design includes four inlet ports consisting of one sample port (Luer interface) and three mini Luer interfaces for fluidic support of e.g. washing buffer, substrate and enzyme solution. The sample can be applied via a special, sealable sampling vessel with integrated female Luer interface. Thereby also pre-anaytical contamination of the environment can be provided. Other reagents that are required for analysis will be stored off chip.

  6. Metrological tests of a 200 L calibration source for HPGE detector systems for assay of radioactive waste drums.

    PubMed

    Boshkova, T; Mitev, K

    2016-03-01

    In this work we present test procedures, approval criteria and results from two metrological inspections of a certified large volume (152)Eu source (drum about 200L) intended for calibration of HPGe gamma assay systems used for activity measurement of radioactive waste drums. The aim of the inspections was to prove the stability of the calibration source during its working life. The large volume source was designed and produced in 2007. It consists of 448 identical sealed radioactive sources (modules) apportioned in 32 transparent plastic tubes which were placed in a wooden matrix which filled the drum. During the inspections the modules were subjected to tests for verification of their certified characteristics. The results show a perfect compliance with the NIST basic guidelines for the properties of a radioactive certified reference material (CRM) and demonstrate the stability of the large volume CRM-drum after 7 years of operation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Overview of the improvement of the ring-stage survival assay-a novel phenotypic assay for the detection of artemisinin-resistant Plasmodium falciparum.

    PubMed

    Zhang, Jie; Feng, Guo-Hua; Zou, Chun-Yan; Su, Pin-Can; Liu, Huai-E; Yang, Zhao-Qing

    2017-11-18

    Artemisinin resistance in Plasmodium falciparum threatens the remarkable efficacy of artemisinin-based combination therapies worldwide. Thus, greater insight into the resistance mechanism using monitoring tools is essential. The ring-stage survival assay is used for phenotyping artemisinin-resistance or decreased artemisinin sensitivity. Here, we review the progress of this measurement assay and explore its limitations and potential applications.

  8. Who needs environmental monitoring?

    Treesearch

    Gary M. Lovett; Douglas A. Burns; Charles T. Driscoll; Jennifer C. Jenkins; Myron J. Mitchell; Lindsey Rustad; James B. Shanley; Gene E. Likens; Richard Haeuber

    2007-01-01

    Environmental monitoring is often criticized as being unscientific, too expensive, and wasteful. While some monitoring studies do suffer from these problems, there are also many highly successful long-term monitoring programs that have provided important scientific advances and crucial information for environmental policy. Here, we discuss the characteristics of...

  9. Waste isolation and contaminant migration - Tools and techniques for monitoring the saturated zone-unsaturated zone-plant-atmosphere continuum

    USGS Publications Warehouse

    Andraski, Brian J.; Stonestrom, David A.; Nicholson, T.J.; Arlt, H.D.

    2011-01-01

    In 1976 the U.S. Geological Survey (USGS) began studies of unsaturated zone hydrology next to the Nation’s first commercial disposal facility for low-level radioactive waste (LLRW) near Beatty, NV. Recognizing the need for long-term data collection, the USGS in 1983 established research management areas in the vicinity of the waste-burial facility through agreements with the Bureau of Land Management and the State of Nevada. Within this framework, the Amargosa Desert Research Site (ADRS; http://nevada.usgs.gov/adrs/) is serving as a field laboratory for the sustained study of water-, gas-, and contaminant-transport processes, and the development of models and methods to characterize flow and transport. The research is built on multiple lines of data that include: micrometeorology; evapotranspiration; plant metrics; soil and sediment properties; unsaturated-zone moisture, temperature, and gas composition; geology and geophysics; and groundwater. Contaminant data include tritium, radiocarbon, volatile-organic compounds (VOCs), and elemental mercury. Presented here is a summary of monitoring tools and techniques that are being applied in studies of waste isolation and contaminant migration.

  10. Non-radioactive TRF assay modifications to improve telomeric DNA detection efficiency in plants

    PubMed Central

    Nigmatullina, Liliia R.; Sharipova, Margarita R.; Shakirov, Eugene V.

    2016-01-01

    The length of telomeric DNA is often considered a cellular biomarker of aging and general health status. Several telomere length measuring assays have been developed, of which the most common is the Telomere Restriction Fragment (TRF) analysis, which typically involves the use of radioactively labeled oligonucleotide probes. While highly effective, this method potentially poses substantial health concerns and generates radioactive waste. Digoxigenin (DIG) alternatives to radioactive probes have been developed and used successfully in a number of assays. Here we optimize the DIG protocol to measure telomere length in the model plant Arabidopsis thaliana and present evidence that this approach can be used successfully to efficiently and accurately measure telomere length in plants. Specifically, hybridization temperature of 42 °C instead of the typical 55 °C appears to generate stronger signals. In addition, DIG incorporation at 5′-end instead of 3′-end of the labeled oligonucleotide greatly enhances signal. We conclude that non-radioactive TRF assays can be as efficient as radioactive methods in detecting and measuring telomere length in plants, making this assay suitable for medical and research laboratories unable to utilize radioactivity due to hazardous waste disposal and safety concerns. PMID:28133587

  11. From Space to the Patient: A New Cytokine Release Assay to Monitor the Immune Status of HIV Infected Patients and Sepsis Patients

    NASA Technical Reports Server (NTRS)

    Kaufmann, I.; Draenert, R.; Gruber, M.; Feuerecker, M.; Crucian, B. E.; Mehta, S. L.; Roider, J.; Pierson, D. L.; Briegel, J. M.; Schelling, G.; hide

    2013-01-01

    Monitoring of humans either in the healthy men under extreme environmental stress like space flight, in human immunodeficiency virus (HIV) infected patients or in sepsis is of critical importance with regard to the timing of adequate therapeutic (counter-)measures. The in vivo skin delayed-type hypersensitivity test (DTH) served for many years as a tool to evaluate cell mediated immunity. However, this standardised in vivo test was removed from the market in 2002 due to the risk of antigen stabilization. To the best of our knowledge an alternative test as monitoring tool to determine cell mediated immunity is not available so far. For this purpose we tested a new alternative assay using elements of the skin DTH which is based on an ex vivo cytokine release from whole blood and asked if it is suitable and applicable to monitor immune changes in HIV infected patients and in patients with septic shock.

  12. Low level waste management: a compilation of models and monitoring techniques. Volume 1

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mosier, J.E.; Fowler, J.R.; Barton, C.J.

    1980-04-01

    In support of the National Low-Level Waste (LLW) Management Research and Development Program being carried out at Oak Ridge National Laboratory, Science Applications, Inc., conducted a survey of models and monitoring techniques associated with the transport of radionuclides and other chemical species from LLW burial sites. As a result of this survey, approximately 350 models were identified. For each model the purpose and a brief description are presented. To the extent possible, a point of contact and reference material are identified. The models are organized into six technical categories: atmospheric transport, dosimetry, food chain, groundwater transport, soil transport, and surfacemore » water transport. About 4% of the models identified covered other aspects of LLW management and are placed in a miscellaneous category. A preliminary assessment of all these models was performed to determine their ability to analyze the transport of other chemical species. The models that appeared to be applicable are identified. A brief survey of the state-of-the-art techniques employed to monitor LLW burial sites is also presented, along with a very brief discussion of up-to-date burial techniques.« less

  13. Enhancing biogas production from anaerobic biodegradation of the organic fraction of municipal solid waste through leachate blending and recirculation.

    PubMed

    Nair, Arjun; Sartaj, Majid; Kennedy, Kevin; Coelho, Nuno M G

    2014-10-01

    Leachate recirculation has a profound advantage on biodegradation of the organic fraction of municipal solid waste in landfills. Mature leachate from older sections of landfills (>10 years) and young leachate were blended and added to organic fraction of municipal solid waste in a series of biomethane potential assay experiments with different mixing ratios of mature and young leachate and their effect on biogas production was monitored. The improvement in biogas production was in the range of 19%-41% depending on the ratio of mixing old and new leachate. The results are conclusive that the biogas generation could be improved by blending the old and new leachate in a bioreactor landfill system as compared with a conventional system employed in bioreactor landfills today for recirculating the same age leachate. © The Author(s) 2014.

  14. SITE GENERATED RADIOLOGICAL WASTE HANDLING SYSTEM DESCRIPTION DOCUMENT

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    S. C. Khamankar

    2000-06-20

    The Site Generated Radiological Waste Handling System handles radioactive waste products that are generated at the geologic repository operations area. The waste is collected, treated if required, packaged for shipment, and shipped to a disposal site. Waste streams include low-level waste (LLW) in solid and liquid forms, as-well-as mixed waste that contains hazardous and radioactive constituents. Liquid LLW is segregated into two streams, non-recyclable and recyclable. The non-recyclable stream may contain detergents or other non-hazardous cleaning agents and is packaged for shipment. The recyclable stream is treated to recycle a large portion of the water while the remaining concentrated wastemore » is packaged for shipment; this greatly reduces the volume of waste requiring disposal. There will be no liquid LLW discharge. Solid LLW consists of wet solids such as ion exchange resins and filter cartridges, as-well-as dry active waste such as tools, protective clothing, and poly bags. Solids will be sorted, volume reduced, and packaged for shipment. The generation of mixed waste at the Monitored Geologic Repository (MGR) is not planned; however, if it does come into existence, it will be collected and packaged for disposal at its point of occurrence, temporarily staged, then shipped to government-approved off-site facilities for disposal. The Site Generated Radiological Waste Handling System has equipment located in both the Waste Treatment Building (WTB) and in the Waste Handling Building (WHB). All types of liquid and solid LLW are processed in the WTB, while wet solid waste from the Pool Water Treatment and Cooling System is packaged where received in the WHB. There is no installed hardware for mixed waste. The Site Generated Radiological Waste Handling System receives waste from locations where water is used for decontamination functions. In most cases the water is piped back to the WTB for processing. The WTB and WHB provide staging areas for storing and

  15. Real-time monitoring system for improving corona electrostatic separation in the process of recovering waste printed circuit boards.

    PubMed

    Li, Jia; Zhou, Quan; Xu, Zhenming

    2014-12-01

    Although corona electrostatic separation is successfully used in recycling waste printed circuit boards in industrial applications, there are problems that cannot be resolved completely, such as nonmetal particle aggregation and spark discharge. Both of these problems damage the process of separation and are not easy to identify during the process of separation in industrial applications. This paper provides a systematic study on a real-time monitoring system. Weight monitoring systems were established to continuously monitor the separation process. A Virtual Instrumentation program written by LabVIEW was utilized to sample and analyse the mass increment of the middling product. It includes four modules: historical data storage, steady-state analysis, data computing and alarm. Three kinds of operating conditions were used to verify the applicability of the monitoring system. It was found that the system achieved the goal of monitoring during the separation process and realized the function of real-time analysis of the received data. The system also gave comprehensible feedback on the accidents of material blockages in the feed inlet and high-voltage spark discharge. With the warning function of the alarm system, the whole monitoring system could save the human cost and help the new technology to be more easily applied in industry. © The Author(s) 2014.

  16. Clinical evaluation of the COBAS Amplicor HBV monitor test for measuring serum HBV DNA and comparison with the Quantiplex branched DNA signal amplification assay in Taiwan.

    PubMed

    Dai, C-Y; Yu, M-L; Chen, S-C; Lin, Z-Y; Hsieh, M-Y; Wang, L-Y; Tsai, J-F; Chuang, W-L; Chang, W-Y

    2004-02-01

    To evaluate the performance characteristics and clinical usefulness of the COBAS Amplicor HBV monitor (COBAS-AM) test in Taiwan and to examine its correlation with the Quantiplex branched DNA signal amplification (bDNA) assay for measuring serum hepatitis B virus (HBV) DNA concentrations. HBV DNA was measured by the COBAS-AM test in 149 sera from chronic HBV infected patients that had previously been analysed by the bDNA assay. The COBAS-AM test showed good reproducibility, with acceptable intra-assay and interassay coefficients of variation (1.6% and 0.9%, respectively) and good linearity (r2=0.98). The overall sensitivity of the COBAS-AM test was significantly higher than that of the bDNA assay (95.3% v 83.2%): 69.6% of samples with HBV DNA below the detection limit of the bDNA assay could be measured by the COBAS-AM test. There was a significant correlation between the results of the two assays (r=0.901; p<0.0001). On average, the results derived from the COBAS-AM test were 0.55 log lower than those of the bDNA assay. HBV DNA concentrations were significantly higher among HBV e antigen (HBeAg) positive patients than negative ones, and higher among patients with abnormal alanine aminotransferase (ALT) concentrations than those with normal ALT concentrations (p=0.0003). The COBAS-AM assay, more sensitive in HBeAg negative samples than the bDNA assay, can effectively measure HBV DNA concentrations in Taiwanese patients. HBV DNA values measured by the COBAS-AM test and bDNA assay correlate significantly.

  17. MONITORING OF INCINERATOR EMISSIONS

    EPA Science Inventory

    Monitoring of Incinerator Emissions is a chapter to be included in a book entitled Hazardous Waste Incineration, edited by A. Sarofim and D. Pershing, and published by John Wiley and Sons. he chapter describes stack sampling and analysis procedures in use on hazardous waste incin...

  18. Gamma neutron assay method and apparatus

    DOEpatents

    Cole, J.D.; Aryaeinejad, R.; Greenwood, R.C.

    1995-01-03

    The gamma neutron assay technique is an alternative method to standard safeguards techniques for the identification and assaying of special nuclear materials in a field or laboratory environment, as a tool for dismantlement and destruction of nuclear weapons, and to determine the isotopic ratios for a blend-down program on uranium. It is capable of determining the isotopic ratios of fissionable material from the spontaneous or induced fission of a sample to within approximately 0.5%. This is based upon the prompt coincidence relationships that occur in the fission process and the proton conservation and quasi-conservation of nuclear mass (A) that exists between the two fission fragments. The system is used in both passive (without an external neutron source) and active (with an external neutron source) mode. The apparatus consists of an array of neutron and gamma-ray detectors electronically connected to determine coincident events. The method can also be used to assay radioactive waste which contains fissile material, even in the presence of a high background radiation field. 7 figures.

  19. Gamma neutron assay method and apparatus

    DOEpatents

    Cole, Jerald D.; Aryaeinejad, Rahmat; Greenwood, Reginald C.

    1995-01-01

    The gamma neutron assay technique is an alternative method to standard safeguards techniques for the identification and assaying of special nuclear materials in a field or laboratory environment, as a tool for dismantlement and destruction of nuclear weapons, and to determine the isotopic ratios for a blend-down program on uranium. It is capable of determining the isotopic ratios of fissionable material from the spontaneous or induced fission of a sample to within approximately 0.5%. This is based upon the prompt coincidence relationships that occur in the fission process and the proton conservation and quasi-conservation of nuclear mass (A) that exists between the two fission fragments. The system is used in both passive (without an external neutron source and active (with an external neutron source) mode. The apparatus consists of an array of neutron and gamma-ray detectors electronically connected to determine coincident events. The method can also be used to assay radioactive waste which contains fissile material, even in the presence of a high background radiation field.

  20. Evaluation of the 5 and 8 pH point titration methods for monitoring anaerobic digesters treating solid waste.

    PubMed

    Vannecke, T P W; Lampens, D R A; Ekama, G A; Volcke, E I P

    2015-01-01

    Simple titration methods certainly deserve consideration for on-site routine monitoring of volatile fatty acid (VFA) concentration and alkalinity during anaerobic digestion (AD), because of their simplicity, speed and cost-effectiveness. In this study, the 5 and 8 pH point titration methods for measuring the VFA concentration and carbonate system alkalinity (H2CO3*-alkalinity) were assessed and compared. For this purpose, synthetic solutions with known H2CO3*-alkalinity and VFA concentration as well as samples from anaerobic digesters treating three different kind of solid wastes were analysed. The results of these two related titration methods were verified with photometric and high-pressure liquid chromatography measurements. It was shown that photometric measurements lead to overestimations of the VFA concentration in the case of coloured samples. In contrast, the 5 pH point titration method provides an accurate estimation of the VFA concentration, clearly corresponding with the true value. Concerning the H2CO3*-alkalinity, the most accurate and precise estimations, showing very similar results for repeated measurements, were obtained using the 8 pH point titration. Overall, it was concluded that the 5 pH point titration method is the preferred method for the practical monitoring of AD of solid wastes due to its robustness, cost efficiency and user-friendliness.

  1. Monitoring the performances of a real scale municipal solid waste composting and a biodrying facility using respiration activity indices.

    PubMed

    Evangelou, Alexandros; Gerassimidou, Spyridoula; Mavrakis, Nikitas; Komilis, Dimitrios

    2016-05-01

    Objective of the work was to monitor two full-scale commingled municipal solid waste (MSW) mechanical and biological pretreatment (MBT) facilities in Greece, namely a biodrying and a composting facility. Monitoring data from a 1.5-year sampling period is presented, whilst microbial respiration indices were used to monitor the decomposition process and the stability status of the wastes in both facilities during the process. Results showed that in the composting facility, the organic matter reduced by 35 % after 8 weeks of combined composting/curing. Material exiting the biocells had a moisture content of less than 30 % (wb) indicating a moisture limitation during the active composting process. The static respiration indexes indicated that some stabilization occurred during the process, but the final material could not be characterized as stable compost. In the biodrying facility, the initial and final moisture contents were 50 % and less than 20 % wb, respectively, and the biodrying index was equal to 4.1 indicating effective biodrying. Lower heating values at the inlet and outlet were approximately 5.5 and 10 MJ/wet kg, respectively. The organic matter was reduced by 20 % during the process and specifically from a range of 63-77 % dw (inlet) to a range of 61-70 % dw. A significant respiration activity reduction was observed for some of the biodrying samples. A statistically significant correlation among all three respiration activity indices was recorded, with the two oxygen related activity indices (CRI7 and SRI24) observing the highest correlation.

  2. WASTE HANDLING BUILDING ELECTRICAL SYSTEM DESCRIPTION DOCUMENT

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    S.C. Khamamkar

    2000-06-23

    The Waste Handling Building Electrical System performs the function of receiving, distributing, transforming, monitoring, and controlling AC and DC power to all waste handling building electrical loads. The system distributes normal electrical power to support all loads that are within the Waste Handling Building (WHB). The system also generates and distributes emergency power to support designated emergency loads within the WHB within specified time limits. The system provides the capability to transfer between normal and emergency power. The system provides emergency power via independent and physically separated distribution feeds from the normal supply. The designated emergency electrical equipment will bemore » designed to operate during and after design basis events (DBEs). The system also provides lighting, grounding, and lightning protection for the Waste Handling Building. The system is located in the Waste Handling Building System. The system consists of a diesel generator, power distribution cables, transformers, switch gear, motor controllers, power panel boards, lighting panel boards, lighting equipment, lightning protection equipment, control cabling, and grounding system. Emergency power is generated with a diesel generator located in a QL-2 structure and connected to the QL-2 bus. The Waste Handling Building Electrical System distributes and controls primary power to acceptable industry standards, and with a dependability compatible with waste handling building reliability objectives for non-safety electrical loads. It also generates and distributes emergency power to the designated emergency loads. The Waste Handling Building Electrical System receives power from the Site Electrical Power System. The primary material handling power interfaces include the Carrier/Cask Handling System, Canister Transfer System, Assembly Transfer System, Waste Package Remediation System, and Disposal Container Handling Systems. The system interfaces with the MGR

  3. Waste Management with Earth Observation Technologies

    NASA Astrophysics Data System (ADS)

    Margarit, Gerard; Tabasco, A.

    2010-05-01

    The range of applications where Earth Observation (EO) can be useful has been notably increased due to the maturity reached in the adopted technology and techniques. In most of the cases, EO provides a manner to remotely monitor particular variables and parameters with a more efficient usage of the available resources. Typical examples are environmental (forest, marine, resources…) monitoring, precision farming, security and surveillance (land, maritime…) and risk / disaster management (subsidence, volcanoes…). In this context, this paper presents a methodology to monitor waste disposal sites with EO. In particular, the explored technology is Interferometric Synthetic Aperture Radar (InSAR), which applies the interferometric concept to SAR images. SAR is an advanced radar concept able to acquire 2D coherent microwave reflectivity images for large scenes (tens of thousands kilometres) with fine resolution (< 1 m). The main product of InSAR is Digital Elevation Models (DEM) that provide key information about the tri-dimensional configuration of a scene, that is, a height map of the scene. In practice, this represents an alternative way to obtain the same information than in-situ altimetry can provide. In the case of waste management, InSAR has been used to evaluate the potentiality of EO to monitor the disposed volume along a specific range of time. This activity has been developed in collaboration with the Agència de Resídus de Catalunya (ARC) (The Waste Agency of Catalonia), Spain, in the framework of a pilot project. The motivation comes from the new law promoted by the regional Government that taxes the volume of disposed waste. This law put ARC in duty to control that the real volume matches the numbers provided by the waste processing firms so that they can not commit illegal actions. Right now, this task is performed with in-situ altimetry. But despite of the accurate results, this option is completely inefficient and limits the numbers of polls that

  4. WASTE HANDLING BUILDING VENTILATION SYSTEM DESCRIPTION DOCUMENT

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    P.A. Kumar

    2000-06-21

    The Waste Handling Building Ventilation System provides heating, ventilation, and air conditioning (HVAC) for the contaminated, potentially contaminated, and uncontaminated areas of the Monitored Geologic Repository's (MGR) Waste Handling Building (WHB). In the uncontaminated areas, the non-confinement area ventilation system maintains the proper environmental conditions for equipment operation and personnel comfort. In the contaminated and potentially contaminated areas, in addition to maintaining the proper environmental conditions for equipment operation and personnel comfort, the contamination confinement area ventilation system directs potentially contaminated air away from personnel in the WHB and confines the contamination within high-efficiency particulate air (HEPA) filtration units. Themore » contamination confinement areas ventilation system creates airflow paths and pressure zones to minimize the potential for spreading contamination within the building. The contamination confinement ventilation system also protects the environment and the public by limiting airborne releases of radioactive or other hazardous contaminants from the WHB. The Waste Handling Building Ventilation System is designed to perform its safety functions under accident conditions and other Design Basis Events (DBEs) (such as earthquakes, tornadoes, fires, and loss of the primary electric power). Additional system design features (such as compartmentalization with independent subsystems) limit the potential for cross-contamination within the WHB. The system provides status of important system parameters and equipment operation, and provides audible and/or visual indication of off-normal conditions and equipment failures. The Waste Handling Building Ventilation System confines the radioactive and hazardous material within the building such that the release rates comply with regulatory limits. The system design, operations, and maintenance activities incorporate ALARA (as low as is

  5. Use of viscoelastic haemostatic assay in emergency and elective surgery.

    PubMed

    Yeung, Maximus C F; Tong, Steven Y T; Tong, Paul Y W; Cheung, Billy H H; Ng, Joanne Y W; Leung, Gilberto K K

    2015-02-01

    To review the current evidence for the use of viscoelastic haemostatic assays in different surgical settings including trauma, cardiac surgery, liver transplantation, as well as the monitoring of antiplatelet agents and anticoagulants prior to surgery. PubMed database. Key words for the literature search were "thromboelastography" or "ROTEM" in combination with "trauma", "antiplatelet", "cardiac surgery", "liver transplantation" or "anticoagulants". Original and major review articles related to the use of viscoelastic haemostatic assays. Haemostatic function is a critical factor determining patient outcomes in emergency or elective surgery. The increasing use of antiplatelet agents and anticoagulants has potentially increased the risks of haemorrhages and the need for transfusion. Conventional coagulation tests have limitations in detecting haemostatic dysfunctions in subgroups of patients and are largely ineffective in diagnosing hyperfibrinolysis. The viscoelastic haemostatic assays are potentially useful point-of-care tools that provide information on clot formation, clot strength, and fibrinolysis, as well as to guide goal-directed transfusion and antifibrinolytic therapy. They may also be used to monitor antiplatelet and anticoagulant therapy. However, standardisation of techniques and reference ranges is required before these tests can be widely used in different clinical settings. Viscoelastic haemostatic assays, as compared with conventional coagulation tests, are better for detecting coagulopathy and are the only tests that can provide rapid diagnosis of hyperfibrinolysis. Goal-directed administration of blood products based on the results of viscoelastic haemostatic assays was associated with reduction in allogeneic blood product transfusions in trauma, cardiac surgery, and liver transplantation cases. However, there is currently no evidence to support the routine use of viscoelastic haemostatic assays for monitoring platelet function prior to surgery.

  6. Nuclear waste

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Not Available

    1991-09-01

    Radioactive waste is mounting at U.S. nuclear power plants at a rate of more than 2,000 metric tons a year. Pursuant to statute and anticipating that a geologic repository would be available in 1998, the Department of Energy (DOE) entered into disposal contracts with nuclear utilities. Now, however, DOE does not expect the repository to be ready before 2010. For this reason, DOE does not want to develop a facility for monitored retrievable storage (MRS) by 1998. This book is concerned about how best to store the waste until a repository is available, congressional requesters asked GAO to review themore » alternatives of continued storage at utilities' reactor sites or transferring waste to an MRS facility, GAO assessed the likelihood of an MRSA facility operating by 1998, legal implications if DOE is not able to take delivery of wastes in 1998, propriety of using the Nuclear Waste Fund-from which DOE's waste program costs are paid-to pay utilities for on-site storage capacity added after 1998, ability of utilities to store their waste on-site until a repository is operating, and relative costs and safety of the two storage alternatives.« less

  7. Annual Progress Report on the Development of Waste Tank Leak Monitoring and Detection and Mitigation Activities in Support of M-45-08

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    DEFIGH PRICE, C.

    2000-09-25

    Milestone M-45-09E of the Hanford Federal Facility Agreement and Consent Order (Tri-Party Agreement or TPA) [TPA 1996] requires submittal of an annual progress report on the development of waste tank leak detection, monitoring, and mitigation (LDMM) activities associated with the retrieval of waste from single-shell tanks (SSTs). This report details progress for fiscal year 2000, building on the current LDMM strategy and including discussion of technologies, applications, cost, schedule, and technical data. The report also includes discussion of demonstrations conducted and recommendations for additional testing. Tri-Party Agreement Milestones M-45-08A and M-45-08B required design and demonstration of LDMM systems for initialmore » retrieval of SST waste. These specific milestones have recently been deleted as part of the M-45-00A change package. Future LDMM development work has been incorporated into specific technology demonstration milestones and SST waste retrieval milestones in the M-45-03 and M-45-05 milestone series.« less

  8. The role of interferon gamma release assays in the monitoring of response to anti-tuberculosis treatment in children.

    PubMed

    Shaik, Junaid; Pillay, Manormoney; Jeena, Prakash

    2014-09-01

    Successful control of childhood TB requires early diagnosis, effective chemotherapy and a method of evaluating the response to therapy. Identification of suitable biomarkers that predict the response to anti-TB therapy may allow the duration of treatment to be shortened. The majority of biomarker studies in paediatric TB have focused on the role of T cell-based interferon-gamma (IFN-γ) release assays (IGRAs) in the diagnosis of either latent or active disease. Little has been published on the role of IGRAs in the monitoring response to therapy in children. We reviewed the available literature to ascertain the value of IGRAs in the monitoring of response to anti-TB therapy in children. We explored the results of the few studies that have investigated the role of IGRAs as markers of response to anti-TB treatment in children. We conclude that the role of IGRAs as surrogate markers appears promising. Robust clinical trials are, however, needed to entrench the value of IGRAs as surrogate biomarkers of response to anti-TB therapy in children. Copyright © 2013 Elsevier Ltd. All rights reserved.

  9. Enhanced surveillance strategies for detecting and monitoring chronic wasting disease in free-ranging cervids

    USGS Publications Warehouse

    Walsh, Daniel P.

    2012-01-01

    The purpose of this document is to provide wildlife management agencies with the foundation upon which they can build scientifically rigorous and cost-effective surveillance and monitoring programs for chronic wasting disease (CWD) or refine their existing programs. The first chapter provides an overview of potential demographic and spatial risk factors of susceptible wildlife populations that may be exploited for CWD surveillance and monitoring. The information contained in this chapter explores historic as well as recent developments in our understanding of CWD disease dynamics. It also contains many literature references for readers who may desire a more thorough review of the topics or CWD in general. The second chapter examines methods for enhancing efforts to detect CWD on the landscape where it is not presently known to exist and focuses on the efficiency and cost-effectiveness of the surveillance program. Specifically, it describes the means of exploiting current knowledge of demographic and spatial risk factors, as described in the first chapter, through a two-stage surveillance scheme that utilizes traditional design-based sampling approaches and novel statistical methods to incorporate information about the attributes of the landscape, environment, populations and individual animals into CWD surveillance activities. By accounting for these attributes, efficiencies can be gained and cost-savings can be realized. The final chapter is unique in relation to the first two chapters. Its focus is on designing programs to monitor CWD once it is discovered within a jurisdiction. Unlike the prior chapters that are more detailed or prescriptive, this chapter by design is considerably more general because providing comprehensive direction for creating monitoring programs for jurisdictions without consideration of their monitoring goals, sociopolitical constraints, or their biological systems, is not possible. Therefore, the authors draw upon their collective

  10. A review on technologies and their usage in solid waste monitoring and management systems: Issues and challenges.

    PubMed

    Hannan, M A; Abdulla Al Mamun, Md; Hussain, Aini; Basri, Hassan; Begum, R A

    2015-09-01

    In the backdrop of prompt advancement, information and communication technology (ICT) has become an inevitable part to plan and design of modern solid waste management (SWM) systems. This study presents a critical review of the existing ICTs and their usage in SWM systems to unfold the issues and challenges towards using integrated technologies based system. To plan, monitor, collect and manage solid waste, the ICTs are divided into four categories such as spatial technologies, identification technologies, data acquisition technologies and data communication technologies. The ICT based SWM systems classified in this paper are based on the first three technologies while the forth one is employed by almost every systems. This review may guide the reader about the basics of available ICTs and their application in SWM to facilitate the search for planning and design of a sustainable new system. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Comparative evaluation of the Cobas Amplicor HIV-1 Monitor Ultrasensitive Test, the new Cobas AmpliPrep/Cobas Amplicor HIV-1 Monitor Ultrasensitive Test and the Versant HIV RNA 3.0 assays for quantitation of HIV-1 RNA in plasma samples.

    PubMed

    Berger, Annemarie; Scherzed, Lina; Stürmer, Martin; Preiser, Wolfgang; Doerr, Hans Wilhelm; Rabenau, Holger Felix

    2005-05-01

    There are several commercially available assays for the quantitation of HIV RNA. A new automated specimen preparation system, the Cobas AmpliPrep, was developed to automate this last part of the PCR. We compared the results obtained by the Roche Cobas Amplicor HIV-1 Monitor Ultrasensitive Test (MCA, manual sample preparation) with those by the Versant HIV-1 RNA 3.0 assay (bDNA). Secondly we compared the MCA with the new Cobas AmpliPrep/Cobas Amplicor HIV Monitor Ultrasensitive Test (CAP/CA, automated specimen preparation) by investigating clinical patient samples and a panel of HIV-1 non-B subtypes. Furthermore, we assessed the assay throughput and workflow (especially hands-on time) for all three assays. Seventy-two percent of the 140 investigated patient samples gave concordant results in the bDNA and MCA assays. The MCA values were regularly higher than the bDNA values. One sample was detected only by the MCA within the linear range of quantification. In contrast, 38 samples with results <50 copies/ml in the MCA showed in the bDNA results between 51 and 1644 copies/ml (mean value 74 copies/ml); 21 of these specimens were shown to have detectable HIV RNA < 50 copies/ml in the MCA assay. The overall agreement between the MCA and the CAP/CA was 94.3% (551/584). The quantification results showed significant correlation, although the CAP/CA generated values slightly lower than those generated by the manual procedure. We found that the CAP/CA produced comparable results with the MCA test in a panel of HIV-1 non-B subtypes. All three assays showed comparable results. The bDNA provides a high sample throughput without the need of full automation. The new CAP/CA provides reliable test results with no HIV-subtype specific influence and releases time for other works in the laboratory; thus it is suitable for routine diagnostic PCR.

  12. Comparative Performance of Three Viral Load Assays on Human Immunodeficiency Virus Type 1 (HIV-1) Isolates Representing Group M (Subtypes A to G) and Group O: LCx HIV RNA Quantitative, AMPLICOR HIV-1 MONITOR Version 1.5, and Quantiplex HIV-1 RNA Version 3.0

    PubMed Central

    Swanson, Priscilla; Soriano, Vincent; Devare, Sushil G.; Hackett, John

    2001-01-01

    The performance of the LCx HIV RNA Quantitative (LCx HIV), AMPLICOR HIV-1 MONITOR version 1.5 (MONITOR v1.5), and Quantiplex HIV-1 RNA version 3.0 (bDNA v3.0) viral load assays was evaluated with 39 viral isolates (3 A, 7 B, 6 C, 4 D, 8 E, 4 F, 1 G, 4 mosaic, and 2 group O). Quantitation across the assay dynamic ranges was assessed using serial fivefold dilutions of the viruses. In addition, sequences of gag-encoded p24 (gag p24), pol-encoded integrase, and env-encoded gp41 were analyzed to assign group and subtype and to assess nucleotide mismatches at primer and probe binding sites. For group M isolates, quantification was highly correlated among all three assays. In contrast, only the LCx HIV assay reliably quantified group O isolates. The bDNA v3.0 assay detected but consistently underquantified group O viruses, whereas the MONITOR v1.5 test failed to detect group O viruses. Analysis of target regions revealed fewer primer or probe mismatches in the LCx HIV assay than in the MONITOR v1.5 test. Consistent with the high level of nucleotide conservation is the ability of the LCx HIV assay to quantify efficiently human immunodeficiency virus type 1 group M and the genetically diverse group O. PMID:11230396

  13. Performance Assessment of Human and Cattle Associated Quantitative Real-time PCR Assays - slides

    EPA Science Inventory

    The presentation overview is (1) Single laboratory performance assessment of human- and cattle associated PCR assays and (2) A Field Study: Evaluation of two human fecal waste management practices in Ohio watershed.

  14. Data analytics approach to create waste generation profiles for waste management and collection.

    PubMed

    Niska, Harri; Serkkola, Ari

    2018-04-30

    Extensive monitoring data on waste generation is increasingly collected in order to implement cost-efficient and sustainable waste management operations. In addition, geospatial data from different registries of the society are opening for free usage. Novel data analytics approaches can be built on the top of the data to produce more detailed, and in-time waste generation information for the basis of waste management and collection. In this paper, a data-based approach based on the self-organizing map (SOM) and the k-means algorithm is developed for creating a set of waste generation type profiles. The approach is demonstrated using the extensive container-level waste weighting data collected in the metropolitan area of Helsinki, Finland. The results obtained highlight the potential of advanced data analytic approaches in producing more detailed waste generation information e.g. for the basis of tailored feedback services for waste producers and the planning and optimization of waste collection and recycling. Copyright © 2018 Elsevier Ltd. All rights reserved.

  15. Indonesia municiple solid waste life cycle and environmental monitoring: current situation, before and future challenges

    NASA Astrophysics Data System (ADS)

    Susmono

    2017-03-01

    Indonesia is a big country with circa 250 million population, with more than 500 Local Governments and they are going to improve their municiple solid waste dumping method from Open Dumping to Sanitary Landfill (SLF) and to promote Reduce-Reuse-Recycling (3R) since many years ago, and it is strengthened by issuing of Solid Waste Management Act No.18/2008, MSW Government Regulation No.12/2012 and other regulations which are issued by Central Government and Local Governments. During “Water and Sanitation Decade 1980-1990” through “Integrated Urban Infrastructures Development Program” some pilot project such as 30 units of 3R station were developed in the urban areas, and modified or simplification of SLF call Controlled Landfill (CLF) were implemented. In the year of 2002 about 45 units of composting pilot projects were developed under “Western Java Environmental Management Project”, and the result was notified that some of them are not sustain because many aspects. At the beginning of 2007 until now, some pilot projects of 3R were continued in some cities and since 2011 some Waste Banks are growing fast. In the year of 2014 was recorded that of 70 % of 3Rs in Java Island well developed (2014, Directorate of Environment Sanitation Report), and in the year of 2012 was recorded that development of Communal Waste Banks were growing fast during two months from 400 units to 800 units (2012, Ministry of Environment report), now more Communal Waste Banks all ready exist. After the last overview monitoring activity by Ministry of Environment and JICA (2008), because of lack of data is very difficult to give current accurate information of Municiple Solid Waste Handling in Indonesia. Nevertheless some innovation are developed because of impact of many pilot projects, Adipura City Cleanest Competition among Local Governments and growing of the spirit of autonomous policy of Local Governments, but some Local Governments still dependence on Central Government support

  16. An enzyme-coupled continuous fluorescence assay for farnesyl diphosphate synthases

    PubMed Central

    Dozier, Jonathan K; Distefano, Mark D

    2012-01-01

    Farnesyl diphosphate synthase (FDPS) catalyzes the conversion of isopentenyl diphosphate and dimethylallyl diphosphate to farnesyl diphosphate, a crucial metabolic intermediate in the synthesis of cholesterol, ubiquinone and prenylated proteins; consequently, much effort has gone into developing inhibitors that target FDPS. Currently most FDPS assays use either radiolabeled substrates and are discontinuous, or monitor pyrophosphate release and not farnesyl diphosphate (FPP) creation. Here we report the development of a continuous coupled enzyme assay for FDPS activity that involves the subsequent incorporation of the FPP product of that reaction into a peptide via the action of protein farnesyltransferase (PFTase). By using a dansylated peptide whose fluorescence quantum yield increases upon farnesylation, the rate of FDPS-catalyzed FPP production can be measured. We show that this assay is more sensitive than existing coupled assays, that it can be used to conveniently monitor FDPS activity in a 96-well plate format and that it can reproduce IC50 values for several previously reported FDPS inhibitors. This new method offers a simple, safe and continuous method to assay FDPS activity that should greatly facilitate the screening of inhibitors of this important target. PMID:22085443

  17. Optical assay for biotechnology and clinical diagnosis.

    PubMed

    Moczko, Ewa; Cauchi, Michael; Turner, Claire; Meglinski, Igor; Piletsky, Sergey

    2011-08-01

    In this paper, we present an optical diagnostic assay consisting of a mixture of environmental-sensitive fluorescent dyes combined with multivariate data analysis for quantitative and qualitative examination of biological and clinical samples. The performance of the assay is based on the analysis of spectrum of the selected fluorescent dyes with the operational principle similar to electronic nose and electronic tongue systems. This approach has been successfully applied for monitoring of growing cell cultures and identification of gastrointestinal diseases in humans.

  18. Proteomics: from hypothesis to quantitative assay on a single platform. Guidelines for developing MRM assays using ion trap mass spectrometers.

    PubMed

    Han, Bomie; Higgs, Richard E

    2008-09-01

    High-throughput HPLC-mass spectrometry (HPLC-MS) is routinely used to profile biological samples for potential protein markers of disease, drug efficacy and toxicity. The discovery technology has advanced to the point where translating hypotheses from proteomic profiling studies into clinical use is the bottleneck to realizing the full potential of these approaches. The first step in this translation is the development and analytical validation of a higher throughput assay with improved sensitivity and selectivity relative to typical profiling assays. Multiple reaction monitoring (MRM) assays are an attractive approach for this stage of biomarker development given their improved sensitivity and specificity, the speed at which the assays can be developed and the quantitative nature of the assay. While the profiling assays are performed with ion trap mass spectrometers, MRM assays are traditionally developed in quadrupole-based mass spectrometers. Development of MRM assays from the same instrument used in the profiling analysis enables a seamless and rapid transition from hypothesis generation to validation. This report provides guidelines for rapidly developing an MRM assay using the same mass spectrometry platform used for profiling experiments (typically ion traps) and reviews methodological and analytical validation considerations. The analytical validation guidelines presented are drawn from existing practices on immunological assays and are applicable to any mass spectrometry platform technology.

  19. Biofiltration of composting gases using different municipal solid waste-pruning residue composts: monitoring by using an electronic nose.

    PubMed

    López, R; Cabeza, I O; Giráldez, I; Díaz, M J

    2011-09-01

    The concentration of volatile organic compounds (VOCs) during the composting of kitchen waste and pruning residues, and the abatement of VOCs by different compost biofilters was studied. VOCs removal efficiencies greater than 90% were obtained using composts of municipal solid waste (MSW) or MSW-pruning residue as biofilter material. An electronic nose identified qualitative differences among the biofilter output gases at very low concentrations of VOCs. These differences were related to compost constituents, compost particle size (2-7 or 7-20mm), and a combination of both factors. The total concentration of VOCs determined by a photoionization analyser and inferred from electronic nose data sets were correlated over an ample range of concentrations of VOCs, showing that these techniques could be specially adapted for the monitoring of these processes. Copyright © 2011 Elsevier Ltd. All rights reserved.

  20. Impedance-based cellular assay technologies: recent advances, future promise.

    PubMed

    McGuinness, Ryan

    2007-10-01

    Cell-based assays are continuing to grow in importance in the drug discovery workflow. Their early introduction holds the promise of limiting attrition in the later, more costly phases of the process. This article reviews recent advances in the development of impedance technologies for label-free cell-based assays. These systems are capable of monitoring endogenous receptor activation, and thus generate more physiologically relevant measures of pharmacological endpoints. Primary cells can be investigated as well, thus producing disease relevant information. Label-free assays significantly decrease assay development efforts and avoid many complications inherent in recombinant readout systems. Impedance-based systems have great potential to advance the utility of cell-based assays as they are applied to drug discovery and pharmacology.

  1. Immune monitoring of clinical trials with biotherapies.

    PubMed

    Whiteside, Theresa L

    2008-01-01

    Immune monitoring of biotherapy clinical trials has undergone a considerable change in recent years. Technical advances together with new insights into molecular immunology have ushered a new genre of assays into immune monitoring. Single-cell assays, multiplex profiling, and signaling molecule detection have replaced formerly used bulk assays, such as proliferation or cytotoxicity. The emphasis on immune cell functions and quantitation of antigen-specific T cells has been playing a major role in attempts to establish correlations between therapy-induced alterations in immune responses and clinical endpoints. However, this has been an elusive goal to achieve, and there is a special need for improving the quality of serial monitoring to ensure that it adequately and reliably measures changes induced by administered biotherapy. In this respect, monitoring performed in specialized reference laboratories operating as good laboratory practice (GLP) facilities and strengthening of interactions between the clinical investigator, the clinical immunologist, and the biostatistician are crucial for successful use of immune monitoring in clinical studies.

  2. ER Operations Installation of Three FLUTe Soil-Vapor Monitoring Wells (MWL-SV03 MWL-SV04 and MWL-SV05) at the Mixed Waste Landfill.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Copland, John Robin

    2014-09-01

    This installation report describes the May through July 2014 drilling activities performed for the installation of three multi-port soil-vapor monitoring wells (MWL-SV03, MWL-SV04, and MWL-SV05) at the Mixed Waste Landfill (MWL), which is located at Sandia National Laboratories, New Mexico (SNL/NM). SNL/NM is managed and operated by Sandia Corporation (Sandia), a wholly owned subsidiary of Lockheed Martin Corporation, for the U.S. Department of Energy (DOE)/National Nuclear Security Administration. The MWL is designated as Solid Waste Management Unit (SWMU) 76 and is located in Technical Area (TA) III (Figure 1-1). The locations of the three soil-vapor monitoring wells (MWL-SV03, MWL-SV04, andmore » MWL-SV05) are shown in Figure 1-2« less

  3. [Shellfish monitoring system for paralytic shellfish toxins using enzyme-linked immunosorbent assay].

    PubMed

    Shinozaki, Takashi; Watanabe, Ryuichi; Kawatsu, Kentaro; Sakurada, Kiyonari; Takahi, Shinya; Ueno, Ken-ichi; Matsushima, Ryoji; Suzuki, Toshiyuki

    2013-01-01

    We investigated the applicability of enzyme-linked immunosorbent assay (PSP-ELISA) using a monoclonal antibody against paralytic shellfish toxins (PST) for screening oysters collected at several coastal areas in Kumamoto prefecture, Japan. Oysters collected between 2007 and 2010 were analyzed by PSP-ELISA. As an alternative calibrant, a naturally contaminated oyster extract was used to quantify toxins in the oyster samples. The toxicity of the calibrant oyster extract determined by the official testing method, mouse bioassay (MBA), was 4 MU/g. Oyster samples collected over 3 years showed a similar toxin profile to the alternative standard, resulting in good agreement between the PSP-ELISA and the MBA. The PSP-ELISA method was better than the MBA in terms of sensitivity, indicating that it may be useful for earlier warning of contamination of oysters by PST in the distinct coastal areas. To use the PSP-ELISA as a screening method prior to MBA, we finally set a screening level at 2 MU/g PSP-ELISA for oyster monitoring in Kumamoto prefecture. We confirmed that there were on samples exceeding the quarantine level (4 MU/g) in MBA among samples quantified as below the screening level by the PSP-ELISA. It was concluded that the use of PSP-ELISA could reduce the numbers of animals needed for MBA testing.

  4. Monitour: Tracking global routes of electronic waste.

    PubMed

    Lee, David; Offenhuber, Dietmar; Duarte, Fábio; Biderman, Assaf; Ratti, Carlo

    2018-02-01

    Many nations seek to control or prevent the inflow of waste electronic and electrical equipment, but such flows are difficult to track due to undocumented, often illegal global trade in e-waste. We apply wireless GPS location trackers to this problem, detecting potential cases of non-compliant recycling operations in the United States as well as the global trajectories of exported e-waste. By planting hidden trackers inside discarded computer monitors and printers, we tracked dozens of devices being sent overseas to various ports in Asia, flows likely unreported in official trade data. We discuss how location tracking enables new ways to monitor, regulate, and enforce rules on the international movement of hazardous electronic waste materials, and the limitations of such methods. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Robotics for mixed waste operations, demonstration description

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ward, C.R.

    The Department of Energy (DOE) Office of Technology Development (OTD) is developing technology to aid in the cleanup of DOE sites. Included in the OTD program are the Robotics Technology Development Program and the Mixed Waste Integrated Program. These two programs are working together to provide technology for the cleanup of mixed waste, which is waste that has both radioactive and hazardous constituents. There are over 240,000 cubic meters of mixed low level waste accumulated at DOE sites and the cleanup is expected to generate about 900,000 cubic meters of mixed low level waste over the next five years. Thismore » waste must be monitored during storage and then treated and disposed of in a cost effective manner acceptable to regulators and the states involved. The Robotics Technology Development Program is developing robotics technology to make these tasks safer, better, faster and cheaper through the Mixed Waste Operations team. This technology will also apply to treatment of transuranic waste. The demonstration at the Savannah River Site on November 2-4, 1993, showed the progress of this technology by DOE, universities and industry over the previous year. Robotics technology for the handling, characterization and treatment of mixed waste as well robotics technology for monitoring of stored waste was demonstrated. It was shown that robotics technology can make future waste storage and waste treatment facilities better, faster, safer and cheaper.« less

  6. Active waste-injection systems in Florida, 1976

    USGS Publications Warehouse

    Vecchioli, John; McKenzie, D.J.; Pascale, C.A.; Wilson, W.E.

    1979-01-01

    As of the end of 1976, seven systems were injecting liquid wastes into Florida 's subsurface environment at a combined average rate of 15 million gallons per day. This report presents for each of these systems information on the kind and amount of waste injected and type of pretreatment, construction characteristics of the injection and monitor wells, type of test and monitoring data available, and brief discussion of any operational problems experienced. (Kosco-USGS)

  7. A FLUORESCENCE BASED ASSAY FOR DNA DAMAGE INDUCED BY STYRENE OXIDE

    EPA Science Inventory

    A rapid and simple assay to detect DNA damage to calf thymus DNA caused by styrene oxide (SO) is reported. This assay is based on changes observed in the melting and annealing behavior of the damaged DNA. The melting annealing process was monitored using a fluorescence indicat...

  8. Validation of an LC-MS/MS assay to simultaneously monitor the intracellular active metabolites of tenofovir, emtricitabine, and lamivudine in dried blood spots.

    PubMed

    Schauer, Amanda P; Sykes, Craig; Cottrell, Mackenzie L; Prince, Heather; Kashuba, Angela D M

    2018-02-05

    The ability to monitor adherence to antiretroviral therapy is critical for the interpretation of outcomes from clinical studies of HIV, and for optimizing patient care. The antiretrovirals tenofovir (TFV), emtricitabine (FTC), and lamivudine (3TC) are commonly included in drug regimens for HIV prevention and treatment. The active form of the drugs tenofovir diphosphate (TFVdp), emtricitabine triphosphate (FTCtp), and lamivudine triphosphate (3TCtp) are found intracellularly in erythrocytes and peripheral blood mononuclear cells (PBMCs). The ability to collect and analyze dried blood spot (DBS) samples is an attractive alternative to PBMC sampling in many resource limited settings. We developed and validated an assay to quantify all three intracellular metabolites over the range of 100-25000 fmol/sample. This assay utilizes a simple protein precipitation/liquid-liquid extraction of a single 3-mm DBS punch (from a Whatman 903 Protein Saver card) with isotopically labeled 13 C 5 -TFVdp included as the internal standard. Following extraction, samples are analyzed by anion exchange chromatography on a Thermo Biobasic AX 5μm column with detection by electrospray ionization in the positive mode on a AB Sciex API-5000 triple quadrupole mass spectrometer with a total run time of 8min. The assay was linear over the entire range (R 2 >0.996). The assay was accurate (inter-assay%bias within ±3.0%) and precise (inter-assay % CV≤9.8%). The assay was also reproducible from multiple punches within a spot as well as punches from separate blood spots. Stability was established at room temperature for 3days, and at -80°C for up to 63days. Clinical samples were analyzed from subjects on Truvada ® , Stribild ® , Descovy ® , and Triumeq ® regimens and intracellular metabolites were detected in all samples as expected, indicating the assay performs well for all current formulations of TFV, FTC, and 3TC. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Comet Assay in Cancer Chemoprevention.

    PubMed

    Santoro, Raffaela; Ferraiuolo, Maria; Morgano, Gian Paolo; Muti, Paola; Strano, Sabrina

    2016-01-01

    The comet assay can be useful in monitoring DNA damage in single cells caused by exposure to genotoxic agents, such as those causing air, water, and soil pollution (e.g., pesticides, dioxins, electromagnetic fields) and chemo- and radiotherapy in cancer patients, or in the assessment of genoprotective effects of chemopreventive molecules. Therefore, it has particular importance in the fields of pharmacology and toxicology, and in both environmental and human biomonitoring. It allows the detection of single strand breaks as well as double-strand breaks and can be used in both normal and cancer cells. Here we describe the alkali method for comet assay, which allows to detect both single- and double-strand DNA breaks.

  10. Recent Advances in Point-of-Access Water Quality Monitoring

    NASA Astrophysics Data System (ADS)

    Korostynska, O.; Arshak, K.; Velusamy, V.; Arshak, A.; Vaseashta, Ashok

    Clean water is one of our most valuable natural resources. In addition to providing safe drinking water it assures functional ecosystems that support fisheries and recreation. Human population growth and its associated increased demands on water pose risks to maintaining acceptable water quality. It is vital to assess source waters and the aquatic systems that receive inputs from industrial waste and sewage treatment plants, storm water systems, and runoff from urban and agricultural lands. Rapid and confident assessments of aquatic resources form the basis for sound environmental management. Current methods engaged in tracing the presence of various bacteria in water employ bulky laboratory equipment and are time consuming. Thus, real-time water quality monitoring is essential for National and International Health and Safety. Environmental water monitoring includes measurements of physical characteristics (e.g. pH, temperature, conductivity), chemical parameters (e.g. oxygen, alkalinity, nitrogen and phosphorus compounds), and abundance of certain biological taxa. Monitoring could also include assays of biological activity such as alkaline phosphatase, tests for toxins such as microcystins and direct measurements of pollutants such as heavy metals or hydrocarbons. Real time detection can significantly reduce the level of damage and also the cost to remedy the problem. This paper presents overview of state-of-the-art methods and devices used for point-of-access water quality monitoring and suggest further developments in this area.

  11. Evaluation of the Abbott ARCHITECT Toxo IgM assay.

    PubMed

    Sickinger, Eva; Braun, Hans-Bertram; Praast, Gerald; Stieler, Myriam; Gundlach, Cordelia; Birkenbach, Claudia; Prostko, John; Palafox, Mary Ann; Frias, Edwin; Hsu, Stephen; Matias, Matthew; Pucci, Dominick; Hausmann, Michael; Sagel, Ulrich; Smith, Darwin

    2009-07-01

    Development of the ARCHITECT Toxo IgM assay has been done to assist the clinician in acute Toxoplasma gondii infection detection, especially in pregnant women. Its use, in conjunction with ARCHITECT Toxo IgG and Toxo Avidity assays, will provide an array of assays particularly useful in the monitoring of pregnant females to determine the risk of maternal transmission of the parasite. Specificity results from 2 testing sites, using populations of pregnant females, hospital patients, and blood donors, demonstrated that the assay has an overall resolved relative specificity of 99.89% (confidence interval, 99.68-99.98%). Relative specificity for pregnant female specimens was 99.95% (n = 2031). Excellent seroconversion sensitivity was observed for the ARCHITECT Toxo IgM assay, which was similar to the Abbott AxSYM Toxo IgM assay (Abbott Laboratories, Abbott Park, IL). In more than 90% of the panels tested, the 1st bleed detected in the serial bleeds was the same for both assays.

  12. Functional requirements for Waste Area Grouping 6 Monitoring Station 3 upgrade

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fiuzat, A.A.

    1991-06-01

    The surface Waste Area Grouping (WAG)6 at ORNL may undergo many changes in the future as part of the closure activities. Changes in surface characteristics will cause changes in runoff characteristics of the MS 3 watershed. Appropriate assumptions are to be made in this project regarding the future surface conditions of the watershed. The extent to which the assumed surface conditions will affect the project objectives (defined in Section 1.0) is to be identified. The purpose of this FRD is to establish performance requirements for MS 3 consisting of the following: (1) The expected range of discharges passing through themore » station and the required accuracy levels for discharge measurements. (2) The equipment required to measure the expected discharges at the required accuracy levels. (3) The sampling requirements for monitoring water quality. (4) The hydraulic requirements for the discharge conveyance structure to be located under the IWMF access road. (5) The design loads to be used for the bridge over the IWMF access road. 12 refs., 5 figs.« less

  13. Programmable flow system for automation of oxygen radical absorbance capacity assay using pyrogallol red for estimation of antioxidant reactivity.

    PubMed

    Ramos, Inês I; Gregório, Bruno J R; Barreiros, Luísa; Magalhães, Luís M; Tóth, Ildikó V; Reis, Salette; Lima, José L F C; Segundo, Marcela A

    2016-04-01

    An automated oxygen radical absorbance capacity (ORAC) method based on programmable flow injection analysis was developed for the assessment of antioxidant reactivity. The method relies on real time spectrophotometric monitoring (540 nm) of pyrogallol red (PGR) bleaching mediated by peroxyl radicals in the presence of antioxidant compounds within the first minute of reaction, providing information about their initial reactivity against this type of radicals. The ORAC-PGR assay under programmable flow format affords a strict control of reaction conditions namely reagent mixing, temperature and reaction timing, which are critical parameters for in situ generation of peroxyl radical from 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH). The influence of reagent concentrations and programmable flow conditions on reaction development was studied, with application of 37.5 µM of PGR and 125 mM of AAPH in the flow cell, guaranteeing first order kinetics towards peroxyl radicals and pseudo-zero order towards PGR. Peroxyl-scavenging reactivity of antioxidants, bioactive compounds and phenolic-rich beverages was estimated employing the proposed methodology. Recovery assays using synthetic saliva provided values of 90 ± 5% for reduced glutathione. Detection limit calculated using the standard antioxidant compound Trolox was 8 μM. RSD values were <3.4 and <4.9%, for intra and inter-assay precision, respectively. Compared to previous batch automated ORAC assays, the developed system also accounted for high sampling frequency (29 h(-1)), low operating costs and low generation of waste. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. 40 CFR 258.55 - Assessment monitoring program.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Section 258.55 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS Ground-Water Monitoring and Corrective Action § 258.55 Assessment... be shown that the removed constituents are not reasonably expected to be in or derived from the waste...

  15. Comparison of Free Light Chain Assays:  Freelite and N Latex in Diagnosis, Monitoring, and Predicting Survival in Light Chain Amyloidosis.

    PubMed

    Mahmood, Shameem; Wassef, Nancy L; Salter, Simon J; Sachchithanantham, Sajitha; Lane, T; Foard, D; Whelan, Carol J; Lachmann, Helen J; Gillmore, Julian D; Hawkins, Philip N; Wechalekar, Ashutosh D

    2016-07-01

    Measurement of serum free light chains (FLCs) is critical in diagnosis, prognosis, and monitoring treatment responses in light chain (AL) amyloidosis. We compare the Freelite assay (polyclonal antibodies to hidden light chain epitopes), which is the current gold standard, with a new assay: a mixture of monoclonal antibodies to light chain epitopes (N Latex). We collected 240 serum samples from 94 consecutive patients with newly diagnosed AL amyloidosis (at least three serial serum samples during the first 6 months) analyzed at the National Amyloidosis Centre, London, from January 2011 to April 2012. Concordance in detecting abnormal light chain components and hematologic response was assessed at 2, 4, and 6 months. The κ and λ clonal light chain involvement was 21% and 79%, respectively, with an abnormal κ/λ ratio or detectable protein in 78.7%. Median κ, λ, and difference in involved and uninvolved FLCs by Freelite and N Latex assays were 17.3 vs 16 mg/L (R(2 ) = 0.91), 48.8 vs 52.6 mg/L (R(2) = 0.52), and 43.2 vs 39.1 mg/L, respectively. Discordant κ/λ ratios at presentation were as follows: 10 of 90 abnormal by Freelite/normal by N Latex and 11 of 90 abnormal by N Latex/normal by Freelite. Both FLC assays show good correlation in detecting the abnormal light chain subtype with discordance in absolute values and thus are not interchangeable. © American Society for Clinical Pathology, 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  16. Health care industries: potential generators of genotoxic waste.

    PubMed

    Sharma, Pratibha; Kumar, Manish; Mathur, N; Singh, A; Bhatnagar, P; Sogani, M

    2013-08-01

    Health care waste includes all the waste generated by health care establishments, research facilities, and laboratories. This constitutes a variety of chemical substances, such as pharmaceuticals, radionuclides, solvents, and disinfectants. Recently, scientists and environmentalists have discovered that wastewater produced by hospitals possesses toxic properties due to various toxic chemicals and pharmaceuticals capable of causing environmental impacts and even lethal effects to organisms in aquatic ecosystems. Many of these compounds resist normal wastewater treatment and end up in surface waters. Besides aquatic organisms, humans can be exposed through drinking water produced from contaminated surface water. Indeed, some of the substances found in wastewaters are genotoxic and are suspected to be potential contributors to certain cancers. The aim of this study was to evaluate the genotoxic and cytotoxic potential of wastewaters from two hospitals and three clinical diagnostic centers located in Jaipur (Rajasthan State), India using the prokaryotic Salmonella mutagenicity assay (Ames assay) and the eukaryotic Saccharomyces cerevisiae respiration inhibition assay. In the Ames assay, untreated wastewaters from both of the health care sectors resulted in significantly increased numbers of revertant colonies up to 1,000-4,050 as measured by the Salmonella typhimurium TA98 and TA100 strains (with and without metabolic activation) after exposure to undiluted samples, which indicated the highly genotoxic nature of these wastewaters. Furthermore, both hospital and diagnostic samples were found to be highly cytotoxic. Effective concentrations at which 20 % (EC20) and 50 % (EC50) inhibition of the respiration rate of the cells occurred ranged between ~0.00 and 0.52 % and between 0.005 and 41.30 % (calculated with the help of the MS excel software XLSTAT 2012.1.01; Addinsoft), respectively, as determined by the S. cerevisiae assay. The results indicated that hospital

  17. Active Sites Environmental Monitoring Program: Mid-FY 1991 report

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ashwood, T.L.; Wickliff, D.S.; Morrissey, C.M.

    1991-10-01

    This report summarizes the activities of the Active Sites Environmental Monitoring Program (ASEMP) from October 1990 through March 1991. The ASEMP was established in 1989 by Solid Waste Operations and the Environmental Sciences Division to provide early detection and performance monitoring at active low-level radioactive waste (LLW) disposal sites in Solid Waste Storage Area (SWSA) 6 and transuranic (TRU) waste storage sites in SWSA 5 as required by chapters II and III of US Department of Energy Order 5820.2A. Monitoring results continue to demonstrate the no LLW is being leached from the storage vaults on the tumulus pads. Loading ofmore » vaults on Tumulus II began during this reporting period and 115 vaults had been loaded by the end of March 1991.« less

  18. Battery operated preconcentration-assisted lateral flow assay.

    PubMed

    Kim, Cheonjung; Yoo, Yong Kyoung; Han, Sung Il; Lee, Junwoo; Lee, Dohwan; Lee, Kyungjae; Hwang, Kyo Seon; Lee, Kyu Hyoung; Chung, Seok; Lee, Jeong Hoon

    2017-07-11

    Paper-based analytical devices (e.g. lateral flow assays) are highly advantageous as portable diagnostic systems owing to their low costs and ease of use. Because of their low sensitivity and detection limits for biomolecules, these devices have several limitations in applications for real-field diagnosis. Here, we demonstrate a paper-based preconcentration enhanced lateral flow assay using a commercial β-hCG-based test. Utilizing a simple 9 V battery operation with a low power consumption of approximately 81 μW, we acquire a 25-fold preconcentration factor, demonstrating a clear sensitivity enhancement in the colorimetric lateral flow assay; consequently, clear colors are observed in a rapid kit test line, which cannot be monitored without preconcentration. This device can also facilitate a semi-quantitative platform using the saturation value and/or color intensity in both paper-based colorimetric assays and smartphone-based diagnostics.

  19. Immunoturbidimetric assay for estimating free light chains of immunoglobulins in urine and serum.

    PubMed Central

    Tillyer, C R; Iqbal, J; Raymond, J; Gore, M; McIlwain, T J

    1991-01-01

    An immunoturbidimetric assay for the assessment of free kappa and lambda light chains of immunoglobulins was developed using a commercial polyclonal antiserum with reactivity towards epitopes on the light chains, which are not expressed when they are bound to heavy chains. The assay, on a centrifugal analyser, is simple and rapid. The limit of detection is 5 mg/l of free light chain, with an assay range of 5-120 mg/l, intrabatch precisions from 1.5-6.4%, and interbatch precisions from 6.5-8.9%. The assay was only slightly less sensitive than colloidal gold staining of cellulose acetate electrophoreses for the detection of Bence-Jones protein in urine. For the serial monitoring of response to chemotherapy in patients with myeloma, the assay correlated well with serum paraprotein estimates obtained by densitometric scanning of Ponceau stained cellulose acetate electrophoreses, but not with serum beta-2 microglobulin measurements, even after correction for the effects of creatinine. These assays may prove to be of use for the monitoring of tumour response in the treatment of Bence-Jones myeloma. PMID:1906071

  20. Qualification of a select one-stage activated partial thromboplastin time-based clotting assay and two chromogenic assays for the post-administration monitoring of nonacog beta pegol.

    PubMed

    Tiefenbacher, S; Bohra, R; Amiral, J; Bowyer, A; Kitchen, S; Lochu, A; Rosén, S; Ezban, M

    2017-10-01

    Essentials Nonacog beta pegol (N9-GP) is an extended half-life, recombinant human factor IX (FIX). One-stage clotting (OSC) and chromogenic FIX activity assays were assessed for N9-GP recovery. OSC STA ® -Cephascreen ® , ROX FIX and BIOPHEN FIX chromogenic assays were qualified for N9-GP. Other extended half-life factor products should be assessed in a similar way prior to approval. Background Nonacog beta pegol (N9-GP) is an extended half-life, glycoPEGylated recombinant human factor IX that is under development for the prophylaxis and treatment of bleeding episodes in hemophilia B patients. Considerable reagent-dependent variability has been observed when one-stage clotting assays are used to measure the recovery of recombinant FIX products, including N9-GP. Objective To qualify select one-stage clotting and chromogenic FIX activity assays for measuring N9-GP recovery. Methods The accuracy and precision of the one-stage clotting assay (with the STA-Cephascreen activated partial thromboplastin [APTT] reagent) and the ROX Factor IX and BIOPHEN Factor IX chromogenic assays for measuring N9-GP recovery were assessed in N9-GP-spiked hemophilia B plasma samples in a systematic manner at three independent sites, with manufacturer-recommended protocols and/or site-specific assay setups, including different instruments. Results For each of the three FIX activity assays qualified on five different reagent-instrument systems, acceptable intra-assay and interassay accuracy and precision, dilution integrity, reagent robustness and freeze-thaw and short-term sample stabilities were demonstrated. The STA-Cephascreen assay showed a limited reportable range at one of the three qualification sites, and the BIOPHEN Factor IX assay showed suspect low-end sensitivity at one of the three qualification sites. An individual laboratory would account for these limitations by adjusting the assay's reportable range; thus, these findings are not considered to impact the respective

  1. Determination of estrogenic potential in waste water without sample extraction.

    PubMed

    Avberšek, Miha; Žegura, Bojana; Filipič, Metka; Uranjek-Ževart, Nataša; Heath, Ester

    2013-09-15

    This study describes the modification of the ER-Calux assay for testing water samples without sample extraction (NE-(ER-Calux) assay). The results are compared to those obtained with ER-Calux assay and a theoretical estrogenic potential obtained by GC-MSD. For spiked tap and waste water samples there was no statistical difference between estrogenic potentials obtained by the three methods. Application of NE-(ER-Calux) to "real" influent and effluents from municipal waste water treatment plants and receiving surface waters found that the NE-(ER-Calux) assay gave higher values compared to ER-Calux assay and GC-MSD. This is explained by the presence of water soluble endocrine agonists that are usually removed during extraction. Intraday dynamics of the estrogenic potential of a WWTP influent and effluent revealed an increase in the estrogenic potential of the influent from 12.9 ng(EEQ)/L in the morning to a peak value of 40.0 ng(EEQ)/L in the afternoon. The estrogenic potential of the effluent was

  2. Effect of leachate recirculation on mesophilic anaerobic digestion of food waste.

    PubMed

    Shahriari, Haleh; Warith, Mostafa; Hamoda, Mohamed; Kennedy, Kevin J

    2012-03-01

    The effects of using untreated leachate for supplemental water addition and liquid recirculation on anaerobic digestion of food waste was evaluated by combining cyclic water recycle operations with batch mesophilic biochemical methane potential (BMP) assays. Cyclic BMP assays indicated that using an appropriate fraction of recycled leachate and fresh make up water can stimulate methanogenic activity and enhance biogas production. Conversely increasing the percentage of recycled leachate in the make up water eventually causes methanogenic inhibition and decrease in the rate of food waste stabilization. The decrease in activity is exacerbated as the number cycles increases. Inhibition is possibly attributed to accumulation and elevated concentrations of ammonia as well as other waste by products in the recycled leachate that inhibit methanogenesis. Copyright © 2011 Elsevier Ltd. All rights reserved.

  3. ORGANOPHOSPHORUS HYDROLASE-BASED ASSAY FOR ORGANOPHOSPHATE PESTICIDES

    EPA Science Inventory

    We report a rapid and versatile Organophosphorus hydrolase (OPH)-based method for measurement of organophosphates. This assay is based on a substrate-dependent change in pH at the local vicinity of the enzyme. The pH change is monitored using fluorescein isothiocyanate (FITC), ...

  4. MRMPlus: an open source quality control and assessment tool for SRM/MRM assay development.

    PubMed

    Aiyetan, Paul; Thomas, Stefani N; Zhang, Zhen; Zhang, Hui

    2015-12-12

    Selected and multiple reaction monitoring involves monitoring a multiplexed assay of proteotypic peptides and associated transitions in mass spectrometry runs. To describe peptide and associated transitions as stable, quantifiable, and reproducible representatives of proteins of interest, experimental and analytical validation is required. However, inadequate and disparate analytical tools and validation methods predispose assay performance measures to errors and inconsistencies. Implemented as a freely available, open-source tool in the platform independent Java programing language, MRMPlus computes analytical measures as recommended recently by the Clinical Proteomics Tumor Analysis Consortium Assay Development Working Group for "Tier 2" assays - that is, non-clinical assays sufficient enough to measure changes due to both biological and experimental perturbations. Computed measures include; limit of detection, lower limit of quantification, linearity, carry-over, partial validation of specificity, and upper limit of quantification. MRMPlus streamlines assay development analytical workflow and therefore minimizes error predisposition. MRMPlus may also be used for performance estimation for targeted assays not described by the Assay Development Working Group. MRMPlus' source codes and compiled binaries can be freely downloaded from https://bitbucket.org/paiyetan/mrmplusgui and https://bitbucket.org/paiyetan/mrmplusgui/downloads respectively.

  5. Characterization of the non-metal fraction of the processed waste printed circuit boards.

    PubMed

    Kumar, Amit; Holuszko, Maria E; Janke, Travis

    2018-05-01

    Electronic waste is one the fastest growing waste streams in the world and waste printed circuit boards (PCB) are the most valuable part of this stream due to the presence of gold, silver, copper, and palladium. The metal present in PCBs is mostly recovered for the market value whereas the nonmetal fractions are often ignored. This research explored the characteristics of the non-metal fraction (NMF) obtained after the processing of milled waste PCBs with a focus on responsible end-of-life solutions, in the form of non-hazardous landfilling or incineration. The NMF was characterized using sizing, assaying, loss on ignition, calorific value measurement, and thermogravimetric analysis (TGA). The result showed that the metal content in the NMF increased with decrease in the particle size for most of the metals except antimony and the result from loss on ignition (LOI) also showed that over 50% of the coarser fraction represented organic matter compared to less than 30% for the finest fraction. The study also showed that after the recovery of metals from the waste PCBs, landfill leaching for most of the metal is reduced below the environmental limits, with lead being the only exception. The lead leachate concentration of 18 mg/L was observed, which requires further treatment prior to landfilling. With an energy value of 16 GJ/t, the NMF could provide high energy recovery if incinerated but 194 mg/kg of hazardous flame retardants present in the NMF might be released if the combustion process is not closely monitored. Copyright © 2018 Elsevier Ltd. All rights reserved.

  6. Clonogenic assay: adherent cells.

    PubMed

    Rafehi, Haloom; Orlowski, Christian; Georgiadis, George T; Ververis, Katherine; El-Osta, Assam; Karagiannis, Tom C

    2011-03-13

    The clonogenic (or colony forming) assay has been established for more than 50 years; the original paper describing the technique was published in 1956. Apart from documenting the method, the initial landmark study generated the first radiation-dose response curve for X-ray irradiated mammalian (HeLa) cells in culture. Basically, the clonogenic assay enables an assessment of the differences in reproductive viability (capacity of cells to produce progeny; i.e. a single cell to form a colony of 50 or more cells) between control untreated cells and cells that have undergone various treatments such as exposure to ionising radiation, various chemical compounds (e.g. cytotoxic agents) or in other cases genetic manipulation. The assay has become the most widely accepted technique in radiation biology and has been widely used for evaluating the radiation sensitivity of different cell lines. Further, the clonogenic assay is commonly used for monitoring the efficacy of radiation modifying compounds and for determining the effects of cytotoxic agents and other anti-cancer therapeutics on colony forming ability, in different cell lines. A typical clonogenic survival experiment using adherent cells lines involves three distinct components, 1) treatment of the cell monolayer in tissue culture flasks, 2) preparation of single cell suspensions and plating an appropriate number of cells in petri dishes and 3) fixing and staining colonies following a relevant incubation period, which could range from 1-3 weeks, depending on the cell line. Here we demonstrate the general procedure for performing the clonogenic assay with adherent cell lines with the use of an immortalized human keratinocyte cell line (FEP-1811). Also, our aims are to describe common features of clonogenic assays including calculation of the plating efficiency and survival fractions after exposure of cells to radiation, and to exemplify modification of radiation-response with the use of a natural antioxidant

  7. Clonogenic Assay: Adherent Cells

    PubMed Central

    Rafehi, Haloom; Orlowski, Christian; Georgiadis, George T.; Ververis, Katherine; El-Osta, Assam; Karagiannis, Tom C.

    2011-01-01

    The clonogenic (or colony forming) assay has been established for more than 50 years; the original paper describing the technique was published in 19561. Apart from documenting the method, the initial landmark study generated the first radiation-dose response curve for X-ray irradiated mammalian (HeLa) cells in culture1. Basically, the clonogenic assay enables an assessment of the differences in reproductive viability (capacity of cells to produce progeny; i.e. a single cell to form a colony of 50 or more cells) between control untreated cells and cells that have undergone various treatments such as exposure to ionising radiation, various chemical compounds (e.g. cytotoxic agents) or in other cases genetic manipulation. The assay has become the most widely accepted technique in radiation biology and has been widely used for evaluating the radiation sensitivity of different cell lines. Further, the clonogenic assay is commonly used for monitoring the efficacy of radiation modifying compounds and for determining the effects of cytotoxic agents and other anti-cancer therapeutics on colony forming ability, in different cell lines. A typical clonogenic survival experiment using adherent cells lines involves three distinct components, 1) treatment of the cell monolayer in tissue culture flasks, 2) preparation of single cell suspensions and plating an appropriate number of cells in petri dishes and 3) fixing and staining colonies following a relevant incubation period, which could range from 1-3 weeks, depending on the cell line. Here we demonstrate the general procedure for performing the clonogenic assay with adherent cell lines with the use of an immortalized human keratinocyte cell line (FEP-1811)2. Also, our aims are to describe common features of clonogenic assays including calculation of the plating efficiency and survival fractions after exposure of cells to radiation, and to exemplify modification of radiation-response with the use of a natural antioxidant

  8. Studies concerning the durability of concrete vaults for intermediate level radioactive waste disposal: Electrochemical monitoring and corrosion aspects

    NASA Astrophysics Data System (ADS)

    Duffó, G. S.; Farina, S. B.; Arva, E. A.; Giordano, C. M.; Lafont, C. J.

    2006-11-01

    The Argentine Atomic Energy Commission (CNEA) is responsible of the development of a management nuclear waste disposal programme. This programme contemplates the design and construction of a facility for the final disposal of intermediate-level radioactive waste. The proposed concept is the near-surface monolithic repository similar to those in operation in El Cabril, Spain. The design of this type of repository is based on the use of multiple, independent and redundant barriers. Since the vault and cover are major components of the engineered barriers, the durability of these concrete structures is an important aspect for the facilities integrity. This work presents a laboratory and field investigation performed for the last 6 years on reinforced concrete specimens, in order to predict the service life of the intermediate level radioactive waste disposal vaults from data obtained from electrochemical techniques. On the other hand, the development of sensors that allow on-line measurements of rebar corrosion potential and corrosion current density; incoming oxygen flow that reaches the metal surface; concrete electrical resistivity and chloride concentration is shown. Those sensors, properly embedded in a new full scale vault (nowadays in construction), will allow the monitoring of the corrosion process of the steel rebars embedded in thestructure.

  9. Capillary electrophoresis-based assay of phosphofructokinase-1.

    PubMed

    Malina, Andrew; Bryant, Sherrisse K; Chang, Simon H; Waldrop, Grover L; Gilman, S Douglass

    2014-02-15

    An assay was developed for phosphofructokinase-1 (PFK-1) using capillary electrophoresis (CE). In the glycolytic pathway, this enzyme catalyzes the rate-limiting step from fructose-6-phosphate and magnesium-bound adenosine triphosphate (Mg-ATP) to fructose-1,6-bisphosphate and magnesium-bound adenosine diphosphate (Mg-ADP). This enzyme has recently become a research target because of the importance of glycolysis in cancer and obesity. The CE assay for PFK-1 is based on the separation and detection by ultraviolet (UV) absorbance at 260 nm of Mg-ATP and Mg-ADP. The separation was enhanced by the addition of Mg²⁺ to the separation buffer. Inhibition studies of PFK-1 by aurintricarboxylic acid and palmitoyl coenzyme A were also performed. An IC₅₀ value was determined for aurintricarboxylic acid, and this value matched values in the literature obtained using coupled spectrophotometric assays. This assay for PFK-1 directly monitors the enzyme-catalyzed reaction, and the CE separation reduces the potential of spectral interference by inhibitors.

  10. Capillary Electrophoresis-Based Assay of Phosphofructokinase-1

    PubMed Central

    Malina, Andrew; Bryant, Sherrisse K.; Chang, Simon H.; Waldrop, Grover L.; Gilman, S. Douglass

    2013-01-01

    An assay was developed for phosphofructokinase-1 (PFK-1) using capillary electrophoresis (CE). In the glycolytic pathway, this enzyme catalyzes the rate-limiting step from fructose-6-phosphate and magnesium-bound adenosine triphosphate (Mg-ATP) to fructose-1,6-bisphosphate and magnesium-bound adenosine diphosphate (Mg-ADP). This enzyme has recently become a research target because of the importance of glycolysis in cancer and obesity. The CE assay for PFK-1 is based on the separation and detection by UV absorbance at 260 nm of Mg-ATP and Mg-ADP. The separation was enhanced by addition of Mg2+ to the separation buffer. Inhibition studies of PFK-1 by aurintricarboxylic acid and palmitoyl coenzyme A were also performed. An IC50 value was determined for aurintricarboxylic acid, and this value matched values in the literature obtained using coupled spectrophotometric assays. This assay for PFK-1 directly monitors the enzyme-catalyzed reaction, and the CE separation reduces the potential of spectral interference by inhibitors. PMID:24444856

  11. Developing immune function assays to monitor fish health in field studies

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rice, C.D.; Kergosien, D.H.; Adams, S.M.

    1995-12-31

    The East Fork Poplar Creek system, a 24km long stream in TN that receives point source discharges of contaminants near its headwaters, was chosen to evaluate a field approach to fish immunotoxicology. Previous studies in this stream have shown that cytochrome P4501A activity, liver somatic indices, macrophage aggregates, and parasitic liver lesions are significantly elevated in sunfish with the degree of impact decreasing with distance from the contaminant source. Red-breasted sunfish were collected between May 23 and June 3 of 1994. Captured fish were anesthetized in MS-222 and processed by two different methods. One group was sacrificed at each samplingmore » station, weights and lengths recorded, and the spleen and anterior kidney tissues removed and placed in buffer on ice. The other group was kept in MS-222 for 2 hr and transported to the laboratory. The spleen and anterior kidney from each fish were then prepared as a single cell suspension and shipped overnight to Mississippi State University. Cells were then washed by centrifugation and resuspended in appropriate media to evaluate PMA-stimulated phagocyte oxidative burst and non-specific cytotoxic cell (NCC) activity against K562 tumor targets. Oxidative burst responses were dramatically suppressed in both groups at stations near the headwaters but returned to reference levels further downstream. There were no differences between treatment groups at each station. NCC activities did not follow gradient-response patterns observed with phagocyte oxidative burst data and there were inconsistent differences between treatment groups at each station. These data show that simple immune function assays, such as phagocyte oxidative burst responses, can be used as an ancillary biomarker in fish health monitoring.« less

  12. Comparison of the Roche COBAS Amplicor Monitor, Roche COBAS Ampliprep/COBAS Taqman and Abbott RealTime Test assays for quantification of hepatitis C virus and HIV RNA.

    PubMed

    Wolff, Dietmar; Gerritzen, Andreas

    2007-01-01

    We have evaluated the performance of two newly developed automated real-time PCR assays, the COBAS Ampliprep/COBAS TaqMan (CAP/CTM) and the Abbott RealTime tests, in the quantification of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) RNA. The widely used semi-automated COBAS Amplicor Monitor (CAM) assay served as the reference test. Several specimens were analyzed, including 102 plasma samples from HCV patients and 109 from HIV patients and 10 samples from negative donors, as well as Quality Control in Molecular Diagnostics (QCMD) and National Institute for Biological Standards and Controls (NIBSC) proficiency program panels. Good correlation was observed among the three assays, with correlation coefficients (R2) of 0.8 (CAM-CAP/CTM), 0.89 (CAM-RealTime) and 0.91 (CAP/CTM-RealTime) for HCV and 0.83 (CAM-RealTime), 0.85 (CAM-CAP/CTM) and 0.89 (CAP/CTM-RealTime) for HIV. The overall concordance for negative/positive results was 100% for HCV and 98% for HIV. All assays were equally able to quantify HCV genotypes 1, 3, 5 and HIV group M (subtypes A-H) and N from QCMD and NIBSC panels. In terms of workflow, the RealTime assay requires more hands-on-time than the CAP/CTM assay. The results indicate that real-time PCR assays can improve the efficiency of end-point PCR tests by better covering viral dynamic ranges and providing higher throughput and automation.

  13. First 25-hydroxyvitamin D assay for general chemistry analyzers.

    PubMed

    Saida, Fakhri B; Chen, Xiaoru; Tran, Kiet; Dou, Chao; Yuan, Chong

    2015-03-01

    25-Hydroxyvitamin D [25(OH)D], the predominant circulating form of vitamin D, is an accurate indicator of the general vitamin D status of an individual. Because vitamin D deficiencies have been linked to several pathologies (including osteoporosis and rickets), accurate monitoring of 25(OH)D levels is becoming increasingly important in clinical settings. Current 25(OH)D assays are either chromatographic or immunoassay-based assays. These assays include HPLC, liquid chromatography-tandem mass spectrometry (LC-MS/MS), enzyme-immunosorbent, immunochemiluminescence, immunofluorescence and radioimmunoassay. All these assays use heterogeneous formats that require phase separation and special instrumentations. In this article, we present an overview of these assays and introduce the first homogeneous assay of 25(OH)D for use on general chemistry analyzers. A special emphasis is put on the unique challenges posed by the 25(OH)D analyte. These challenges include a low detection limit, the dissociation of the analyte from its serum transporter and the inactivation of various binding proteins without phase separation steps.

  14. Kinetic modelling of anaerobic hydrolysis of solid wastes, including disintegration processes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    García-Gen, Santiago; Sousbie, Philippe; Rangaraj, Ganesh

    2015-01-15

    Highlights: • Fractionation of solid wastes into readily and slowly biodegradable fractions. • Kinetic coefficients estimation from mono-digestion batch assays. • Validation of kinetic coefficients with a co-digestion continuous experiment. • Simulation of batch and continuous experiments with an ADM1-based model. - Abstract: A methodology to estimate disintegration and hydrolysis kinetic parameters of solid wastes and validate an ADM1-based anaerobic co-digestion model is presented. Kinetic parameters of the model were calibrated from batch reactor experiments treating individually fruit and vegetable wastes (among other residues) following a new protocol for batch tests. In addition, decoupled disintegration kinetics for readily and slowlymore » biodegradable fractions of solid wastes was considered. Calibrated parameters from batch assays of individual substrates were used to validate the model for a semi-continuous co-digestion operation treating simultaneously 5 fruit and vegetable wastes. The semi-continuous experiment was carried out in a lab-scale CSTR reactor for 15 weeks at organic loading rate ranging between 2.0 and 4.7 g VS/L d. The model (built in Matlab/Simulink) fit to a large extent the experimental results in both batch and semi-continuous mode and served as a powerful tool to simulate the digestion or co-digestion of solid wastes.« less

  15. Transboundary movements of hazardous wastes: the case of toxic waste dumping in Africa.

    PubMed

    Anyinam, C A

    1991-01-01

    Developed and developing countries are in the throes of environmental crisis. The planet earth is increasingly being literally choked by the waste by-products of development. Of major concern, especially to industrialized countries, is the problem of what to do with the millions of tons of waste materials produced each year. Owing to mounting pressure from environmental groups, the "not-in-mu-backyard" movement, the close monitoring of the activities of waste management agents, an increasing paucity of repositories for waste, and the high cost of waste treatment, the search for dumping sites for waste disposal has, in recent years, extended beyond regional and national boundaries. The 1980s have seen several attempts to export hazardous wastes to third world countries. Africa, for example, is gradually becoming the prime hunting ground for waste disposal companies. This article seeks to examine, in the context of the African continent, the sources and destinations of this form of relocation-diffusion of pollution, factors that have contributed to international trade in hazardous wastes between developed and developing countries, the potential problems such exports would bring to African countries, and measures being taken to abolish this form of international trade.

  16. A European multicenter study on the analytical performance of the VERIS HBV assay.

    PubMed

    Braun, Patrick; Delgado, Rafael; Drago, Monica; Fanti, Diana; Fleury, Hervé; Izopet, Jacques; Lombardi, Alessandra; Mancon, Alessandro; Marcos, Maria Angeles; Sauné, Karine; O Shea, Siobhan; Pérez-Rivilla, Alfredo; Ramble, John; Trimoulet, Pascale; Vila, Jordi; Whittaker, Duncan; Artus, Alain; Rhodes, Daniel

    Hepatitis B viral load monitoring is an essential part of managing patients with chronic Hepatits B infection. Beckman Coulter has developed the VERIS HBV Assay for use on the fully automated Beckman Coulter DxN VERIS Molecular Diagnostics System. 1 OBJECTIVES: To evaluate the analytical performance of the VERIS HBV Assay at multiple European virology laboratories. Precision, analytical sensitivity, negative sample performance, linearity and performance with major HBV genotypes/subtypes for the VERIS HBV Assay was evaluated. Precision showed an SD of 0.15 log 10 IU/mL or less for each level tested. Analytical sensitivity determined by probit analysis was between 6.8-8.0 IU/mL. Clinical specificity on 90 unique patient samples was 100.0%. Performance with 754 negative samples demonstrated 100.0% not detected results, and a carryover study showed no cross contamination. Linearity using clinical samples was shown from 1.23-8.23 log 10 IU/mL and the assay detected and showed linearity with major HBV genotypes/subtypes. The VERIS HBV Assay demonstrated comparable analytical performance to other currently marketed assays for HBV DNA monitoring. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Forecasting waste compositions: A case study on plastic waste of electronic display housings.

    PubMed

    Peeters, Jef R; Vanegas, Paul; Kellens, Karel; Wang, Feng; Huisman, Jaco; Dewulf, Wim; Duflou, Joost R

    2015-12-01

    Because of the rapid succession of technological developments, the architecture and material composition of many products used in daily life have drastically changed over the last decades. As a result, well-adjusted recycling technologies need to be developed and installed to cope with these evolutions. This is essential to guarantee continued access to materials and to reduce the ecological impact of our material consumption. However, limited information is currently available on the material composition of arising waste streams and even less on how these waste streams will evolve. Therefore, this paper presents a methodology to forecast trends in the material composition of waste streams. To demonstrate the applicability and value of the proposed methodology, it is applied to forecast the evolution of plastic housing waste from flat panel display (FPD) TVs, FPD monitors, cathode ray tube (CRT) TVs and CRT monitors. The results of the presented forecasts indicate that a wide variety of plastic types and additives, such as flame retardants, are found in housings of similar products. The presented case study demonstrates that the proposed methodology allows the identification of trends in the evolution of the material composition of waste streams. In addition, it is demonstrated that the recycling sector will need to adapt its processes to deal with the increasing complexity of plastics of end-of-life electronic displays while respecting relevant directives. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Mobile site safety review for the transuranic (TRU) waste characterization program

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Not Available

    1996-11-01

    This Safety Review Document (SRD) applies to the Active/Passive Neutron Examination and Assay (APNEA) system installed on a Lockheed Martin Specialty Components, Inc., (Specialty Components) trailer. The APNEA is designed to perform nuclear waste drum assay. The purpose of this document is to describe the safety features of the APNEA system.

  19. A novel assay for discovery and characterization of pro-apoptotic drugs and for monitoring apoptosis in patient sera.

    PubMed

    Bivén, K; Erdal, H; Hägg, M; Ueno, T; Zhou, R; Lynch, M; Rowley, B; Wood, J; Zhang, C; Toi, M; Shoshan, M C; Linder, S

    2003-06-01

    We have developed an apoptosis assay based on measurement of a neoepitope of cytokeratin-18 (CK18-Asp396) exposed after caspase-cleavage and detected by the monoclonal antibody M30. The total amount of caspase-cleaved CK18 which has accumulated in cells and tissue culture media during apoptosis is measured by ELISA. The sensitivity is sufficient for use in the 96-well format to allow high-through-put screening of drug libraries. We here describe strategies allowing classification of pro-apoptotic compounds according to their profiles of induction of apoptosis in the presence of pharmacological inhibitors. The time course of induction of CK18 cleavage can furthermore be used to distinguish structurally similar compounds. We propose that compounds that induce rapid CK18 cleavage have mechanisms of actions distinct from conventional genotoxic and microtubuli-targeting agents, and we present one example of an agent that induces almost immediate mitochondrial depolarization and cytochrome c release. Finally, CK18-Asp396 cleavage products are released from cells in tissue culture, and presumably from tumor cells in vivo. These products can be measured in sera from cancer patients. We present evidence suggesting that it will be possible to use the M30-ELISA assay for measuring chemotherapy-induced apoptosis in patient sera, opening possibilities for monitoring therapy.

  20. Comprehensive comparison of the VERSANT HIV-1 RNA 3.0 (bDNA) and COBAS AMPLICOR HIV-1 MONITOR 1.5 assays on 1,000 clinical specimens.

    PubMed

    Galli, Rick; Merrick, Linda; Friesenhahn, Michel; Ziermann, Rainer

    2005-12-01

    Plasma human immunodeficiency virus type 1 (HIV-1) RNA level is an important parameter for patient management, yet viral load assays from different manufacturers are not standardized. In this study, we evaluated the concordance between test results obtained for 1,000 plasma specimens collected from HIV-1-infected individuals measured with the VERSANT HIV-1 RNA 3.0 assay (bDNA) and the COBAS AMPLICOR HIV-1 MONITOR 1.5 test (PCR). We compared viral load values obtained by each of these assays throughout their dynamic ranges, with particular focus on samples with low viral load (i.e. 50-250 copies/mL), and calculated the estimated distribution of distinct plasma viral load levels for the entire study population modeled from the data observed in the study. We found that these two assays show excellent agreement, with a correlation (R(2)) of 0.957 and a slope of 1.004. The mean difference in viral load values between the two assays was less than 0.10-log(10) throughout the dynamic range and 98.2% of all samples had bDNA and PCR results within 0.5-log(10) of each other, a difference that is within the range considered to be a minimal change in plasma viremia. Moreover, the two assays show very similar results across all assay ranges tested. The estimated prevalence of samples with results <50 copies/mL, 50-250 copies/mL, and 250-500,000 copies/mL were 41.6%, 7.7%, and 49.7%, respectively, by the bDNA assay, and 42.4%, 6.9%, and 50.7%, respectively, by the PCR assay. Based on our findings from 1,000 clinical specimens, we do not see the need to re-establish a baseline value or apply a conversion factor when switching from one assay to the other. Since the majority of our patient population likely is infected with subtype B virus, it is unclear if our findings will apply to other patient populations with a greater incidence of infection with non-B subtypes.

  1. Real time monitoring and quantification of reactive oxygen species in breast cancer cell line MCF-7 by 2',7'-dichlorofluorescin diacetate (DCFDA) assay.

    PubMed

    Figueroa, Daniela; Asaduzzaman, Mohammad; Young, Fiona

    2018-04-07

    The detection of reactive oxygen species (ROS) using 2',7'-dichlorofluorescin diacetate (DCFDA) is commonly performed by a single measurement of fluorescence but this fails to capture a profile of ROS generation over time. This study aimed to develop a real-time monitoring method to increase the utility of the assay, to incorporate cytotoxicity screening and to describe the combined effects of DCFDA and the ROS generator, Ter-butyl hydrogen peroxide (TBHP). Breast cancer MCF-7 cells were loaded with DCFDA (0-50 μM) for 45 min, and then exposed to TBHP (0-50 μM). Fluorescence was recorded according to three different schedules: every hour for 6 h, or once after 6 h or 24 h. Viability was assessed in a crystal violet assay and cell morphology was examined by microscopy. TBHP caused a time and dose-dependent increase in ROS and the magnitude of the fluorescent signal was affected by the loading concentration of DCFDA. Reading the fluorescence every hour for 6 h did not diminish the emission signal. The most sensitive and reliable combination for this ROS assay was 10 μM DCFDA with 25 μM TBHP; since higher concentrations of DCFDA compromised cell viability. In conclusion we adapted a single point ROS assay to enable production of a profile of ROS generation over an extended 6 h period, and related this to cell viability and morphology. Published by Elsevier Inc.

  2. Simulation of municipal solid waste degradation in aerobic and anaerobic bioreactor landfills.

    PubMed

    Patil, Bhagwan Shamrao; C, Agnes Anto; Singh, Devendra Narain

    2017-03-01

    Municipal solid waste generation is huge in growing cities of developing nations such as India, owing to the rapid industrial and population growth. In addition to various methods for treatment and disposal of municipal solid waste (landfills, composting, bio-methanation, incineration and pyrolysis), aerobic/anaerobic bioreactor landfills are gaining popularity for economical and effective disposal of municipal solid waste. However, efficiency of municipal solid waste bioreactor landfills primarily depends on the municipal solid waste decomposition rate, which can be accelerated through monitoring moisture content and temperature by using the frequency domain reflectometry probe and thermocouples, respectively. The present study demonstrates that these landfill physical properties of the heterogeneous municipal solid waste mass can be monitored using these instruments, which facilitates proper scheduling of the leachate recirculation for accelerating the decomposition rate of municipal solid waste.

  3. A Label-Free Continuous Fluorescence-Based Assay for Monitoring Ornithine Decarboxylase Activity with a Synthetic Putrescine Receptor.

    PubMed

    Nilam, Mohamed; Gribbon, Philip; Reinshagen, Jeanette; Cordts, Kathrin; Schwedhelm, Edzard; Nau, Werner M; Hennig, Andreas

    2017-08-01

    Polyamines play an important role in cell growth, differentiation, and cancer development, and the biosynthetic pathway of polyamines is established as a drug target for the treatment of parasitic diseases, neoplasia, and cancer chemoprevention. The key enzyme in polyamine biosynthesis is ornithine decarboxylase (ODC). We report herein an analytical method for the continuous fluorescence monitoring of ODC activity based on the supramolecular receptor cucurbit[6]uril (CB6) and the fluorescent dye trans-4-[4-(dimethylamino)styryl]-1-methylpyridinium iodide (DSMI). CB6 has a significantly higher binding constant to the ODC product putrescine (>10 7 M -1 ) than to the substrate L-ornithine (340 M -1 ). This enables real-time monitoring of the enzymatic reaction through a continuous fluorescence change caused by dye displacement from the macrocycle by the formed product, which allowed a straightforward determination of enzyme kinetic parameters ( k cat = 0.12 s -1 and K M = 24 µM) and inhibition constants of the two ODC inhibitors α-difluoromethylornithine (DFMO) and epigallocatechin gallate (EGCG). The potential for high-throughput screening (HTS) was demonstrated by excellent Z' factors (>0.9) in a microplate reader format, and the sensitivity of the assay is comparable to or better than most established complementary methods, which invariably have the disadvantage of not being compatible with direct implementation and upscaling to HTS format in the drug discovery process.

  4. Targeting Anti-Cancer Active Compounds: Affinity-Based Chromatographic Assays

    PubMed Central

    de Moraes, Marcela Cristina; Cardoso, Carmen Lucia; Seidl, Claudia; Moaddel, Ruin; Cass, Quezia Bezerra

    2016-01-01

    Affinity-based chromatography assays encompass the use of solid supports containing immobilized biological targets to monitor binding events in the isolation , identification and/or characterization of bioactive compounds. This powerful bioanalytical technique allows the screening of potential binders through fast analyses that can be directly performed using isolated substances or complex matrices. An overview of the recent researches in frontal and zonal affinity-based chromatography screening assays, which has been used as a tool in the identification and characterization of new anti-cancer agents, is discussed. In addition, a critical evaluation of the recently emerged ligands fishing assays in complex mixtures is also discussed. PMID:27306095

  5. Monitoring the Durability Performance of Concrete in Nuclear Waste Containment. Technical Progress Report No. 4

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ulm, Franz-Josef

    2000-06-30

    OAK-B135 Monitoring the Durability Performance of Concrete in Nuclear Waste Containment. Technical Progress Report No. 4. The analysis of the effect of cracks on the acceleration of the calcium leaching process of cement-based materials has been pursued. During the last period (Technical Progress Report No 3), we have introduced a modeling accounting for the high diffusivity of fractures in comparison with the weak solid material diffusivity. It has been shown through dimensional and asymptotic analysis that small fractures do not significantly accelerate the material aging process. This important result for the overall structural aging kinetics of containment structure has beenmore » developed in a paper submitted to the international journal ''Transport in Porous Media''.« less

  6. 21 CFR 866.5910 - Quality control material for cystic fibrosis nucleic acid assays.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunological Test Systems § 866.5910 Quality control material for cystic fibrosis nucleic acid assays. (a... cystic fibrosis nucleic acid assays is a device intended to help monitor reliability of a test system by...

  7. Development, Testing and Validation of a Waste Assay System for the Measurement and Characterisation of Active Spent Fuel Element Debris From UK Magnox Reactors - 12533

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mason, John A.; Burke, Kevin J.; Looman, Marc R.

    2012-07-01

    eliminate interferences from radioactive sources and FED in the immediate vicinity of the measurement position. The detector system has been calibrated and high activity radioactive sources of Cs-137, Co-60 and Na-22 have been used to validate the measurement process. The data acquisition and analysis software code has been tested and validated in keeping with the software quality assurance requirements of both ISO:9001-2008 - TICK-IT in the UK and NQA-1. The measurement and analysis system has been comprehensively tested with high activity sources, is flexible and may be applicable to a wide range of remote handled radioactive waste measurement applications. It is due to be installed at Trawsfynydd NPS later this year. This paper describes the Waste Tray Assay System (WTAS) that has been developed for the measurement of Magnox FED waste. The WTAS has been tested with a range of radioactive sources and its operation has been simulated with benchmarked MCNP Monte Carlo calculations. The measurement software has been validated as has the operation of the system for a range of strong radioactive sources. A system based on the design is due for installation and operation in 2012. The system has application to the measurement of Magnox Fuel Element Debris (FED) waste at other Magnox reactor sites. The major design objective of the WTAS that has been achieved is the ability of the assay system to determine the content of Cs-137, and in turn to enable the fissile burden to be assessed using a radionuclide fingerprint, in the presence of higher and highly variable quantities of Co-60, typically from nimonic springs. The approach can be used in other Magnox FED waste configurations where the detector is located above the FED waste sorting tray and where the collimation is fixed below the detector and at a distance above the tray. In this case, which has also been investigated, there are different shielding problems and mechanical support issues. The extensive use of MCNP Monte Carlo

  8. Optical assays based on colloidal inorganic nanoparticles.

    PubMed

    Ghasemi, Amir; Rabiee, Navid; Ahmadi, Sepideh; Hashemzadeh, Shabnam; Lolasi, Farshad; Bozorgomid, Mahnaz; Kalbasi, Alireza; Nasseri, Behzad; Shiralizadeh Dezfuli, Amin; Aref, Amir Reza; Karimi, Mahdi; Hamblin, Michael R

    2018-06-20

    Colloidal inorganic nanoparticles have wide applications in the detection of analytes and in biological assays. A large number of these assays rely on the ability of gold nanoparticles (AuNPs, in the 20 nm diameter size range) to undergo a color change from red to blue upon aggregation. AuNP assays can be based on cross-linking, non-cross linking or unmodified charge-based aggregation. Nucleic acid-based probes, monoclonal antibodies, and molecular-affinity agents can be attached by covalent or non-covalent means. Surface plasmon resonance and SERS techniques can be utilized. Silver NPs also have attractive optical properties (higher extinction coefficient). Combinations of AuNPs and AgNPs in nanocomposites can have additional advantages. Magnetic NPs and ZnO, TiO2 and ZnS as well as insulator NPs including SiO2 can be employed in colorimetric assays, and some can act as peroxidase mimics in catalytic applications. This review covers the synthesis and stabilization of inorganic NPs and their diverse applications in colorimetric and optical assays for analytes related to environmental contamination (metal ions and pesticides), and for early diagnosis and monitoring of diseases, using medically important biomarkers.

  9. Performance evaluation of the Aptima® HCV Quant Dx assay for hepatitis C virus (HCV) RNA detection and quantification in comparison to the Abbott RealTime HCV assay.

    PubMed

    Garbuglia, Anna Rosa; Bibbò, Angela; Sciamanna, Roberta; Pisciotta, Marina; Capobianchi, Maria Rosaria

    2017-07-01

    The Aptima HCV Quant Dx assay (Aptima) is a real-time transcription-mediated amplification assay CE-approved for the diagnosis and monitoring of hepatitis C virus (HCV) infection. Aptima's analytical performance was compared to the Abbott RealTime HCV assay (RealTime) in a clinical routine setting. Overall 295 clinical plasma samples (117 prospective/fresh; 178 retrospective/frozen) from HCV-infected patients were tested in Aptima and RealTime to determine concordance on qualitative and quantitative results. Linearity and precision at low viral loads (VLs; 0.8-3.3LogIU/mL) was tested using dilutions of the 5th WHO standard, in 10 and 20 replicates in the two assays, respectively. The ability to measure different HCV genotypes and accuracy were analyzed using the Seracare EQA panel. Inter-assay agreement for qualitative results (prospective samples) was 88% (kappa=0.78). For the 127 samples with quantitative results in both assays, Aptima yielded on average slightly higher values (by 0.24LogIU/mL; Bland-Altman method) than RealTime. Concordance between assay results was excellent (R=0.98). At low VLs (0.8-3.3LogIU/mL), Aptima demonstrated good linearity and precision, similar to RealTime. Aptima detected and accurately quantified all main HCV genotypes. Aptima demonstrated excellent precision, linearity, and accuracy in all genotypes tested. Good concordance was observed between Aptima and RealTime assays in clinical samples. The performance of the Aptima assay, on the fully automated Panther platform, makes it an excellent candidate for the detection and monitoring of HCV RNA in plasma and serum samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Monitoring household waste recycling centres performance using mean bin weight analyses.

    PubMed

    Maynard, Sarah; Cherrett, Tom; Waterson, Ben

    2009-02-01

    This paper describes a modelling approach used to investigate the significance of key factors (vehicle type, compaction type, site design, temporal effects) in influencing the variability in observed nett amenity bin weights produced by household waste recycling centres (HWRCs). This new method can help to quickly identify sites that are producing significantly lighter bins, enabling detailed back-end analyses to be efficiently targeted and best practice in HWRC operation identified. Tested on weigh ticket data from nine HWRCs across West Sussex, UK, the model suggests that compaction technique, vehicle type, month and site design explained 76% of the variability in the observed nett amenity weights. For each factor, a weighting coefficient was calculated to generate a predicted nett weight for each bin transaction and three sites were subsequently identified as having similar characteristics but returned significantly different mean nett bin weights. Waste and site audits were then conducted at the three sites to try and determine the possible sources of the remaining variability. Significant differences were identified in the proportions of contained waste (bagged), wood, and dry recyclables entering the amenity waste stream, particularly at one site where significantly less contaminated waste and dry recyclables were observed.

  11. Archaeal community structure in leachate and solid waste is correlated to methane generation and volume reduction during biodegradation of municipal solid waste.

    PubMed

    Fei, Xunchang; Zekkos, Dimitrios; Raskin, Lutgarde

    2015-02-01

    Duplicate carefully-characterized municipal solid waste (MSW) specimens were reconstituted with waste constituents obtained from a MSW landfill and biodegraded in large-scale landfill simulators for about a year. Repeatability and relationships between changes in physical, chemical, and microbial characteristics taking place during the biodegradation process were evaluated. Parameters such as rate of change of soluble chemical oxygen demand in the leachate (rsCOD), rate of methane generation (rCH4), rate of specimen volume reduction (rVt), DNA concentration in the leachate, and archaeal community structures in the leachate and solid waste were monitored during operation. The DNA concentration in the leachate was correlated to rCH4 and rVt. The rCH4 was related to rsCOD and rVt when waste biodegradation was intensive. The structures of archaeal communities in the leachate and solid waste of both simulators were very similar and Methanobacteriaceae were the dominant archaeal family throughout the testing period. Monitoring the chemical and microbial characteristics of the leachate was informative of the biodegradation process and volume reduction in the simulators, suggesting that leachate monitoring could be informative of the extent of biodegradation in a full-scale landfill. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Biomonitoring of genotoxicity of industrial fertilizer pollutants in Aiolopus thalassinus (Orthoptera: Acrididae) using alkaline comet assay.

    PubMed

    Abdelfattah, Eman A; Augustyniak, Maria; Yousef, Hesham A

    2017-09-01

    Phosphate fertilizer industry is considered as one of the main sources of environmental pollutants. Besides solid waste products, e.g. phosphates, sulphates, and heavy metals, also atmospheric pollutants, such as hydrofluoric acid fumes (HF), sulphur dioxide (SO 2 ), nitrogen oxides (NO 2 ), and particulate matter with diameter up to 10 μm (PM 10 ) can be dangerous. Genotoxic effect of these pollutants was monitored by assessing the DNA damage using alkaline comet assay on cells from brain, thoracic muscles and gut of Aiolopus thalassinus collected at three sites (A-C) located at 1, 3, and 6 km away from Abu-Zaabal Company for Fertilizers and Chemical Industries. Control site was established 32 km from the source of pollution, at the Cairo University Campus. The level of the DNA damage was significantly higher in insects from polluted sites comparing to that from the control site. A strong negative correlation between percentage of cells with visible DNA damage (% of severed cells) and the distance of the sites from Abu-Zaabal Company was found. The best parameter for monitoring of fertilizer pollutants is % of severed cells. Possible impact of Abu-Zaabal Company (extremely high concentration of phosphates and sulphates in all the polluted sites) on DNA integrity in A. thalassinus tissues was discussed. The potential use of the comet assay as a biomonitoring method of the environmental pollution caused by fertilizer industry was proposed. Specific pollution resulting from the activity of the fertilizer industry can cause comparable adverse effects in the organisms inhabiting areas up to 6 km from the source of contamination. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. A single secreted luciferase-based gene reporter assay.

    PubMed

    Barriscale, Kathy A; O'Sullivan, Sharon A; McCarthy, Tommie V

    2014-05-15

    Promoter analysis typically employs a reporter gene fused to a test promoter combined with a second reporter fused to a control promoter that is used for normalization purposes. However, this approach is not valid when experimental conditions affect the control promoter. We have developed and validated a single secreted luciferase reporter (SSLR) assay for promoter analysis that avoids the use of a control reporter. The approach uses an early level of expression of a secreted luciferase linked to a test promoter as an internal normalization control for subsequent analysis of the same promoter. Comparison of the SSLR assay with the dual luciferase reporter (DLR) assay using HMGCR (3-hydroxy-3-methylglutaryl-coenzyme A reductase) and LDLR (low-density lipoprotein receptor) promoter constructs, which are down-regulated by 25-hydroxycholesterol, show that both assays yield similar results. Comparison of the response of the HMGCR promoter in SSLR transient assays compared very favorably with the response of the same promoter in the stable cell line. Overall, the SSLR assay proved to be a valid alternative to the DLR assay for certain applications and had significant advantages in that measurement of only one luciferase is required and monitoring can be continuous because cell lysis is not necessary. Copyright © 2014 Elsevier Inc. All rights reserved.

  14. Impedance-based cellular assays for regenerative medicine.

    PubMed

    Gamal, W; Wu, H; Underwood, I; Jia, J; Smith, S; Bagnaninchi, P O

    2018-07-05

    Therapies based on regenerative techniques have the potential to radically improve healthcare in the coming years. As a result, there is an emerging need for non-destructive and label-free technologies to assess the quality of engineered tissues and cell-based products prior to their use in the clinic. In parallel, the emerging regenerative medicine industry that aims to produce stem cells and their progeny on a large scale will benefit from moving away from existing destructive biochemical assays towards data-driven automation and control at the industrial scale. Impedance-based cellular assays (IBCA) have emerged as an alternative approach to study stem-cell properties and cumulative studies, reviewed here, have shown their potential to monitor stem-cell renewal, differentiation and maturation. They offer a novel method to non-destructively assess and quality-control stem-cell cultures. In addition, when combined with in vitro disease models they provide complementary insights as label-free phenotypic assays. IBCA provide quantitative and very sensitive results that can easily be automated and up-scaled in multi-well format. When facing the emerging challenge of real-time monitoring of three-dimensional cell culture dielectric spectroscopy and electrical impedance tomography represent viable alternatives to two-dimensional impedance sensing.This article is part of the theme issue 'Designer human tissue: coming to a lab near you'. © 2018 The Author(s).

  15. Alternative Methods for the Detection of Emerging Marine Toxins: Biosensors, Biochemical Assays and Cell-Based Assays

    PubMed Central

    Reverté, Laia; Soliño, Lucía; Carnicer, Olga; Diogène, Jorge; Campàs, Mònica

    2014-01-01

    The emergence of marine toxins in water and seafood may have a considerable impact on public health. Although the tendency in Europe is to consolidate, when possible, official reference methods based on instrumental analysis, the development of alternative or complementary methods providing functional or toxicological information may provide advantages in terms of risk identification, but also low cost, simplicity, ease of use and high-throughput analysis. This article gives an overview of the immunoassays, cell-based assays, receptor-binding assays and biosensors that have been developed for the screening and quantification of emerging marine toxins: palytoxins, ciguatoxins, cyclic imines and tetrodotoxins. Their advantages and limitations are discussed, as well as their possible integration in research and monitoring programs. PMID:25431968

  16. Recommendations for the generation, quantification, storage and handling of peptides used for mass spectrometry-based assays

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hoofnagle, Andrew N.; Whiteaker, Jeffrey R.; Carr, Steven A.

    2015-12-30

    The Clinical Proteomic Tumor Analysis Consortium (1) (CPTAC) of the National Cancer Institute (NCI) is a comprehensive and coordinated effort to accelerate the understanding of the molecular basis of cancer through the application of robust technologies and workflows for the quantitative measurements of proteins. The Assay Development Working Group of the CPTAC Program aims to foster broad uptake of targeted mass spectrometry-based assays employing isotopically labeled peptides for confident assignment and quantification, including multiple reaction monitoring (MRM; also referred to as Selected Reaction Monitoring), parallel reaction monitoring (PRM), and other targeted methods.

  17. Luciferase reporter assay in Drosophila and mammalian tissue culture cells

    PubMed Central

    Yun, Chi

    2014-01-01

    Luciferase reporter gene assays are one of the most common methods for monitoring gene activity. Because of their sensitivity, dynamic range, and lack of endogenous activity, luciferase assays have been particularly useful for functional genomics in cell-based assays, such as RNAi screening. This unit describes delivery of two luciferase reporters with other nucleic acids (siRNA /dsRNA), measurement of the dual luciferase activities, and analysis of data generated. The systematic query of gene function (RNAi) combined with the advances in luminescent technology have made it possible to design powerful whole genome screens to address diverse and significant biological questions. PMID:24652620

  18. Effect of genotypes on the quantification of hepatitis C virus (HCV) RNA in clinical samples using the Amplicor HCV Monitor Test and the Quantiplex HCV RNA 2.0 assay (bDNA).

    PubMed

    Tong, C Y; Hollingsworth, R C; Williams, H; Irving, W L; Gilmore, I T

    1998-07-01

    The Amplicor HCV Monitor test and the Quantiplex HCV RNA 2.0 (bDNA) assay are two commercially available assays for the quantification of hepatitis C virus (HCV) RNA in clinical samples. A direct comparison of the two assays was carried out using sera frozen previously from patients known to be chronically infected with HCV. Overall, 61 samples from 51 patients were tested simultaneously by the two methods: 67% (28/42) of the patients were infected by HCV genotype/serotype 1, 10% (4/42) with type 2, and 24% (10/42) with type 3. When the absolute value from each assay was examined, the Quantiplex assay gave a consistently higher reading and the mean logarithmic difference between the two assays was 1.4 (1.0 in type 1, 2.0 in type 2, and 2.2 in type 3). When analyzed according to genotype, strong correlation was observed between the two assays for type 1 (r = 0.83, 95% CI 0.63-0.93, P < 0.01), but not for nontype 1 samples. Despite the difference in absolute level reported by the two assays, there was a consistent trend of change in HCV RNA concentration by both assays in patients whose consecutive samples were analyzed and the differences between the two assays in consecutive samples were within 0.4 log of each other. The results suggested that with samples containing genotype 1, the Amplicor assay was more sensitive than the Quantiplex assay by about one log. However, the sensitivities of the two assays with nontype 1 samples were much closer probably due to the failure of the Amplicor assay to quantify nontype 1 genotypes effectively.

  19. Clinical application of the Quantiplex HCV RNA 2.0 and Amplicor HCV Monitor assays for quantifying serum hepatitis C virus RNA.

    PubMed

    Yu, M L; Chuang, W L; Chen, S C; Lin, Z Y; Hsieh, M Y; Wang, L Y; Chang, W Y

    1999-11-01

    To compare the performance characteristics and clinical application of two different technologies for quantifying serum hepatitis C virus (HCV) RNA levels. HCV RNA was quantified by Amplicor HCV Monitor assay (Amplicor) and Quantiplex HCV RNA 2.0 assay (bDNA-2) in 119 sera from 107 HCV infected patients. Both assays had similar sensitivity (79.4% for Amplicor; 86.0% for bDNA-2), acceptable coefficients of variation (5.3% in Amplicor; 2.6% in bDNA-2), and good linearity (r2 > or = 0.98). There was a positive correlation between quantification values of both methods (r = 0.683, p < 0.001). The Amplicor values were on an average 1.76 log lower than bDNA-2 results. Male subjects and HCV genotype 1b were significantly associated with higher viral load determined by Amplicor, but not with viral load measured by bDNA-2. In 70 chronic HCV infected patients treated with interferon alfa, mean (SD) pretreatment viral load in 27 complete responders (3.47 (0.84) logs for Amplicor, 5.63 (0.58) for bDNA-2) was significantly lower than in non-responders (4.43 (1.01) logs for Amplicor, 6.10 (0.67) logs for bDNA-2; p < 0.001). Cut off points of 3.9 logs for Amplicor and 5.8 logs for bDNA-2 were determined to be the best for predicting response to interferon alfa, giving acceptable sensitivity (70.4%, 74.1%), specificity (72.1%, 65.1%), and accuracy (71.4%, 68.6%), respectively. Both the Amplicor and bDNA-2 assays are clinically useful methods for HCV RNA quantification and are reliable for predicting the outcome of treatment, despite differences in absolute quantification values and in the correlation between HCV genotypes and viral load.

  20. EFFICIENT MONITORING OF HETEROGENEOUS MEDIA AND ELECTRONIC WASTES

    EPA Science Inventory

    The Agency's Office of Solid Waste and Emergency Response (OSWER) has recently issued improved guidance for the collection of "representative" samples from heterogeneous media. The Technology Innovation Office (TIO) has begun the development of a web-based handbook which advocat...

  1. Anti-Peptide Monoclonal Antibodies Generated for Immuno-Multiple Reaction Monitoring-Mass Spectrometry Assays Have a High Probability of Supporting Western blot and ELISA*

    PubMed Central

    Schoenherr, Regine M.; Saul, Richard G.; Whiteaker, Jeffrey R.; Yan, Ping; Whiteley, Gordon R.; Paulovich, Amanda G.

    2015-01-01

    Immunoaffinity enrichment of peptides coupled to targeted, multiple reaction monitoring-mass spectrometry (immuno-MRM) has recently been developed for quantitative analysis of peptide and protein expression. As part of this technology, antibodies are generated to short, linear, tryptic peptides that are well-suited for detection by mass spectrometry. Despite its favorable analytical performance, a major obstacle to widespread adoption of immuno-MRM is a lack of validated affinity reagents because commercial antibody suppliers are reluctant to commit resources to producing anti-peptide antibodies for immuno-MRM while the market is much larger for conventional technologies, especially Western blotting and ELISA. Part of this reluctance has been the concern that affinity reagents generated to short, linear, tryptic peptide sequences may not perform well in traditional assays that detect full-length proteins. In this study, we test the feasibility and success rates of generating immuno-MRM monoclonal antibodies (mAbs) (targeting tryptic peptide antigens) that are also compatible with conventional, protein-based immuno-affinity technologies. We generated 40 novel, peptide immuno-MRM assays and determined that the cross-over success rates for using immuno-MRM monoclonals for Western blotting is 58% and for ELISA is 43%, which compare favorably to cross-over success rates amongst conventional immunoassay technologies. These success rates could most likely be increased if conventional and immuno-MRM antigen design strategies were combined, and we suggest a workflow for such a comprehensive approach. Additionally, the 40 novel immuno-MRM assays underwent fit-for-purpose analytical validation, and all mAbs and assays have been made available as a resource to the community via the Clinical Proteomic Tumor Analysis Consortium's (CPTAC) Antibody (http://antibodies.cancer.gov) and Assay Portals (http://assays.cancer.gov), respectively. This study also represents the first

  2. 40 CFR 258.51 - Ground-water monitoring systems.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Ground-water monitoring systems. 258... CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS Ground-Water Monitoring and Corrective Action § 258.51 Ground-water monitoring systems. (a) A ground-water monitoring system must be installed that consists of a...

  3. 40 CFR 257.22 - Ground-water monitoring systems.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Ground-water monitoring systems. 257... Waste Disposal Units Ground-Water Monitoring and Corrective Action § 257.22 Ground-water monitoring systems. (a) A ground-water monitoring system must be installed that consists of a sufficient number of...

  4. 40 CFR 258.51 - Ground-water monitoring systems.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 25 2014-07-01 2014-07-01 false Ground-water monitoring systems. 258... CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS Ground-Water Monitoring and Corrective Action § 258.51 Ground-water monitoring systems. (a) A ground-water monitoring system must be installed that consists of a...

  5. 40 CFR 258.51 - Ground-water monitoring systems.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 26 2012-07-01 2011-07-01 true Ground-water monitoring systems. 258.51... FOR MUNICIPAL SOLID WASTE LANDFILLS Ground-Water Monitoring and Corrective Action § 258.51 Ground-water monitoring systems. (a) A ground-water monitoring system must be installed that consists of a...

  6. 40 CFR 257.22 - Ground-water monitoring systems.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 25 2014-07-01 2014-07-01 false Ground-water monitoring systems. 257... Waste Disposal Units Ground-Water Monitoring and Corrective Action § 257.22 Ground-water monitoring systems. (a) A ground-water monitoring system must be installed that consists of a sufficient number of...

  7. Validated electrochemical and chromatographic quantifications of some antibiotic residues in pharmaceutical industrial waste water.

    PubMed

    Ibrahim, Heba K; Abdel-Moety, Mona M; Abdel-Gawad, Sherif A; Al-Ghobashy, Medhat A; Kawy, Mohamed Abdel

    2017-03-01

    Realistic implementation of ion selective electrodes (ISEs) into environmental monitoring programs has always been a challenging task. This could be largely attributed to difficulties in validation of ISE assay results. In this study, the electrochemical response of amoxicillin trihydrate (AMX), ciprofloxacin hydrochloride (CPLX), trimethoprim (TMP), and norfloxacin (NFLX) was studied by the fabrication of sensitive membrane electrodes belonging to two types of ISEs, which are polyvinyl chloride (PVC) membrane electrodes and glassy carbon (GC) electrodes. Linear response for the membrane electrodes was in the concentration range of 10 -5 -10 -2  mol/L. For the PVC membrane electrodes, Nernstian slopes of 55.1, 56.5, 56.5, and 54.0 mV/decade were achieved over a pH 4-8 for AMX, CPLX, and NFLX, respectively, and pH 3-6 for TMP. On the other hand, for GC electrodes, Nernstian slopes of 59.1, 58.2, 57.0, and 58.2 mV/decade were achieved over pH 4-8 for AMX, CPLX, and NFLX, respectively, and pH 3-6 for TMP. In addition to assay validation to international industry standards, the fabricated electrodes were also cross-validated relative to conventional separation techniques; high performance liquid chromatography (HPLC), and thin layer chromatography (TLC)-densitometry. The HPLC assay was applied in concentration range of 0.5-10.0 μg/mL, for all target analytes. The TLC-densitometry was adopted over a concentration range of 0.3-1.0 μg/band, for AMX, and 0.1-0.9 μg/band, for CPLX, NFLX, and TMP. The proposed techniques were successfully applied for quantification of the selected drugs either in pure form or waste water samples obtained from pharmaceutical plants. The actual waste water samples were subjected to solid phase extraction (SPE) for pretreatment prior to the application of chromatographic techniques (HPLC and TLC-densitometry). On the other hand, the fabricated electrodes were successfully applied for quantification of the antibiotic residues in actual

  8. Visible and infrared remote imaging of hazardous waste: A review

    USGS Publications Warehouse

    Slonecker, Terrence; Fisher, Gary B.; Aiello, Danielle P.; Haack, Barry

    2010-01-01

    One of the critical global environmental problems is human and ecological exposure to hazardous wastes from agricultural, industrial, military and mining activities. These wastes often include heavy metals, hydrocarbons and other organic chemicals. Traditional field and laboratory detection and monitoring of these wastes are generally expensive and time consuming. The synoptic perspective of overhead remote imaging can be very useful for the detection and remediation of hazardous wastes. Aerial photography has a long and effective record in waste site evaluations. Aerial photographic archives allow temporal evaluation and change detection by visual interpretation. Multispectral aircraft and satellite systems have been successfully employed in both spectral and morphological analysis of hazardous wastes on the landscape and emerging hyperspectral sensors have permitted determination of the specific contaminants by processing strategies using the tens or hundreds of acquired wavelengths in the solar reflected and/or thermal infrared parts of the electromagnetic spectrum. This paper reviews the literature of remote sensing and overhead imaging in the context of hazardous waste and discusses future monitoring needs and emerging scientific research areas.

  9. 40 CFR 264.99 - Compliance monitoring program.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... be based on a compliance monitoring program developed to meet the requirements of this section. (i... 40 Protection of Environment 26 2011-07-01 2011-07-01 false Compliance monitoring program. 264.99... Releases From Solid Waste Management Units § 264.99 Compliance monitoring program. An owner or operator...

  10. Renilla Luciferase-LC3 Based Reporter Assay for Measuring Autophagic Flux.

    PubMed

    Farkas, T; Jäättelä, M

    2017-01-01

    Macroautophagy (autophagy) is a dynamic intracellular degradation pathway. Monitoring the flux through the autophagy pathway is experimentally challenging but obviously a prerequisite for the proper investigation of the process. Here, we present an indirect autophagy flux assay based on monitoring the degradation of an autophagosome-associated fusion protein Rluc-LC3 by luminescence detection. The method is suitable for screening purposes with a high number of parallel samples and can be used for measurements in cell lysates as well as in living cells. The Rluc-LC3 assay has proven useful for the identification of genes, miRNAs, and small molecules that regulate autophagy flux in mammalian cells. © 2017 Elsevier Inc. All rights reserved.

  11. Aptamer-based assay for monitoring genetic disorder phenylketonuria (PKU).

    PubMed

    Hasanzadeh, Mohammad; Zargami, Amir; Baghban, Hossein Navay; Mokhtarzadeh, Ahad; Shadjou, Nasrin; Mahboob, Soltanali

    2018-05-16

    The genetic disorder phenylketonuria (PKU) is the inability to metabolize phenylalanine because of a lack of the enzyme phenylalanine hydroxylase. Phenylalanine is used to biochemically form proteins, coded for by DNA. The development of an apta-assay for detection of l-Phenylalanine is presented in this work. A highly specific DNA-aptamer, selected to l-Phenylalanine was immobilized onto a gold nanostructure and electrochemical measurements were performed in a solution containing the phosphate buffer solution with physiological pH. We have constructed an aptamer immobilized gold nanostructure mediated, ultrasensitive electrochemical biosensor (Apt/AuNSs/Au electrode) for l-Phenylalanine detection without any additional signal amplification strategy. The aptamer assemble onto the AuNSs makes Apt/AuNSs/Au electrode an excellent platform for the l-Phenylalanine detection in physiological like condition. Differential pulse voltammetry were used for the quantitative l-Phenylalanine detection. The Apt/AuNSs/Au electrode offers an ultrasensitive and selective detection of l-Phenylalanine down to 0.23 μM level with a wide dynamic range from 0.72 μM-6 mM. The aptasensor exhibited excellent selectivity and stability. The real sample analysis was performed by spiking the unprocessed human serum samples with various concentration of l-Phenylalanine and obtained recovery within 2% error value. The sensor is found to be more sensitive than most of the literature reports. The simple and easy way of construction of this apta-assay provides an efficient and promising diagnosis of phenylketonuria. Copyright © 2018. Published by Elsevier B.V.

  12. Safety monitoring of the FBG sensor in respect of radioactivity and deformation measurement of a silo structure for radioactive waste disposal

    NASA Astrophysics Data System (ADS)

    Kim, Ki-Soo; Cho, Seong-Kyu

    2015-07-01

    The FBG sensor has globally been commercialized in various fields that is actively applied in Korea as well. Especially it is widely used as a structural monitoring sensor in civil engineering and construction structures due to its advantages including electrical stability, chemical stability and multiplexing. This report aims to introduce safety inspection of the FBG sensor in respect of radioactivity which has been applied to a silo structure for radioactive waste disposal as an example.

  13. Subsurface injection of liquid waste in Florida, United States of America

    USGS Publications Warehouse

    Vecchioli, John

    1981-01-01

    In 1979, liquid waste was injected into the subsurface of Florida by 10 injection systems at an aggregate average rate of 165,000 m3/d. All the systems inject into carbonate rocks that contain salty water. Extensive precautions are taken in the construction of the injection wells and in the monitoring of their operation to provide assurance that overlying and laterally contiguous freshwater resources do not become contaminated with either the injected waste or the saltwater displaced by the waste. Several concerns relating to the effectiveness of the confining bed above the injection zone for containing the injected wastes have arisen over the years. These concerns accentuate the value of a well-planned and implemented monitoring program from which one can evaluate the potential impact of waste injection on the subsurface environment.

  14. Real-time monitoring of lactate extrusion and glucose consumption of cultured cells using a lab-on-valve system.

    PubMed

    Schulz, Craig M; Scampavia, Louis; Ruzicka, Jaromir

    2002-12-01

    Microsequential injection (microST) provides microfluidic operations that are ideally suited for cellular function studies and as a means of validating targets for drug discovery. MicroSI carried out within the lab-on-valve (LOV) manifold, is an ideal platform for spectroscopic studies on living cells that are grown on microcarrier beads and kept thermostated while their metabolism is probed in real-time. In this paper a microbioreactor is integrated into the LOV manifold allowing measurement of cellular lactate extrusion and glucose consumption rates of a cell culture that is automatically renewed prior to each measurement. Glucose consumption and lactate extrusion are monitored using NAD-linked enzymatic assays. The microSI-LOV setup has demonstrated a linear analysis range of 0.05-1.00 mM for lactate and 0.1-5.6 mM for glucose. These assays were conducted in a serial fashion requiring 3 microL of cellular perfusate and 10 s for glucose determination and 30 s for the lactate assay. Overall waste generated per lactate/glucose assay is < 200 microL. This work was performed using two different transfected hepatocyte cell lines, which adhere to Cytopore microcarrier beads. This novel approach to metabolic screening allows for the rapid evaluation of the effects of dosing cells with chemical agents.

  15. Mass sensing AlN sensors for waste water monitoring

    NASA Astrophysics Data System (ADS)

    Porrazzo, R.; Potter, G.; Lydecker, L.; Foraida, Z.; Gattu, S.; Tokranova, N.; Castracane, J.

    2014-08-01

    Monitoring the presence of nanomaterials in waste water from semiconductor facilities is a critical task for public health organizations. Advanced semiconductor technology allows the fabrication of sensitive piezoelectric-based mass sensors with a detection limit of less than 1.35 ng/cm2 of nanomaterials such as nanoparticles of alumina, amorphous silica, ceria, etc. The interactions between acoustic waves generated by the piezoelectric sensor and nanomaterial mass attached to its surface define the sensing response as a shift in the resonant frequency. In this article the development and characterization of a prototype AlN film bulk acoustic resonator (FBAR) are presented. DC reactive magnetron sputtering was used to create tilted c-axis oriented AlN films to generate shear waves which don't propagate in liquids thus minimizing the acoustic losses. The high acoustic velocity of AlN over quartz allows an increase in resonance frequency in comparison with a quartz crystal microbalance (QCM) and results in a higher frequency shift per mass change, and thus greater sensitivity. The membrane and electrodes were fabricated using state of the art semiconductor technology. The device surface functionalization was performed to demonstrate selectivity towards a specific nanomaterial. As a result, the devices were covered with a "docking" layer that allows the nanomaterials to be selectively attached to the surface. This was achieved using covalent modification of the surface, specifically targeting ZnO nanoparticles. Our functionalization approach was tested using two different types of nanoparticles, and binding specificity was confirmed with various analytical techniques.

  16. 40 CFR 60.2165 - What monitoring equipment must I install and what parameters must I monitor?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... installed in each baghouse compartment or cell. For negative pressure or induced air fabric filters, the bag... test. (g) For waste-burning kilns not equipped with a wet scrubber, in place of hydrogen chloride..., maintain, and operate a continuous emission monitoring system for monitoring hydrogen chloride emissions...

  17. Micro-scale anaerobic digestion of point source components of organic fraction of municipal solid waste

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chanakya, H.N.; Sharma, Isha; Ramachandra, T.V.

    The fermentation characteristics of six specific types of the organic fraction of municipal solid waste (OFMSW) were examined, with an emphasis on properties that are needed when designing plug-flow type anaerobic bioreactors. More specifically, the decomposition patterns of a vegetable (cabbage), fruits (banana and citrus peels), fresh leaf litter of bamboo and teak leaves, and paper (newsprint) waste streams as feedstocks were studied. Individual OFMSW components were placed into nylon mesh bags and subjected to various fermentation periods (solids retention time, SRT) within the inlet of a functioning plug-flow biogas fermentor. These were removed at periodic intervals, and their compositionmore » was analyzed to monitor decomposition rates and changes in chemical composition. Components like cabbage waste, banana peels, and orange peels fermented rapidly both in a plug-flow biogas reactor (PFBR) as well as under a biological methane potential (BMP) assay, while other OFMSW components (leaf litter from bamboo and teak leaves and newsprint) fermented slowly with poor process stability and moderate biodegradation. For fruit and vegetable wastes (FVW), a rapid and efficient removal of pectins is the main cause of rapid disintegration of these feedstocks, which left behind very little compost forming residues (2-5%). Teak and bamboo leaves and newsprint decomposed only to 25-50% in 30 d. These results confirm the potential for volatile fatty acids accumulation in a PFBR's inlet and suggest a modification of the inlet zone or operation of a PFBR with the above feedstocks.« less

  18. Micro-scale anaerobic digestion of point source components of organic fraction of municipal solid waste.

    PubMed

    Chanakya, H N; Sharma, Isha; Ramachandra, T V

    2009-04-01

    The fermentation characteristics of six specific types of the organic fraction of municipal solid waste (OFMSW) were examined, with an emphasis on properties that are needed when designing plug-flow type anaerobic bioreactors. More specifically, the decomposition patterns of a vegetable (cabbage), fruits (banana and citrus peels), fresh leaf litter of bamboo and teak leaves, and paper (newsprint) waste streams as feedstocks were studied. Individual OFMSW components were placed into nylon mesh bags and subjected to various fermentation periods (solids retention time, SRT) within the inlet of a functioning plug-flow biogas fermentor. These were removed at periodic intervals, and their composition was analyzed to monitor decomposition rates and changes in chemical composition. Components like cabbage waste, banana peels, and orange peels fermented rapidly both in a plug-flow biogas reactor (PFBR) as well as under a biological methane potential (BMP) assay, while other OFMSW components (leaf litter from bamboo and teak leaves and newsprint) fermented slowly with poor process stability and moderate biodegradation. For fruit and vegetable wastes (FVW), a rapid and efficient removal of pectins is the main cause of rapid disintegration of these feedstocks, which left behind very little compost forming residues (2-5%). Teak and bamboo leaves and newsprint decomposed only to 25-50% in 30d. These results confirm the potential for volatile fatty acids accumulation in a PFBR's inlet and suggest a modification of the inlet zone or operation of a PFBR with the above feedstocks.

  19. 40 CFR 264.97 - General ground-water monitoring requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 26 2014-07-01 2014-07-01 false General ground-water monitoring requirements. 264.97 Section 264.97 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) STANDARDS FOR OWNERS AND OPERATORS OF HAZARDOUS WASTE TREATMENT, STORAGE, AND DISPOSAL FACILITIES Releases From Solid Waste...

  20. Improved Methods for Capture, Extraction, and Quantitative Assay of Environmental DNA from Asian Bigheaded Carp (Hypophthalmichthys spp.)

    PubMed Central

    Turner, Cameron R.; Miller, Derryl J.; Coyne, Kathryn J.; Corush, Joel

    2014-01-01

    Indirect, non-invasive detection of rare aquatic macrofauna using aqueous environmental DNA (eDNA) is a relatively new approach to population and biodiversity monitoring. As such, the sensitivity of monitoring results to different methods of eDNA capture, extraction, and detection is being investigated in many ecosystems and species. One of the first and largest conservation programs with eDNA-based monitoring as a central instrument focuses on Asian bigheaded carp (Hypophthalmichthys spp.), an invasive fish spreading toward the Laurentian Great Lakes. However, the standard eDNA methods of this program have not advanced since their development in 2010. We developed new, quantitative, and more cost-effective methods and tested them against the standard protocols. In laboratory testing, our new quantitative PCR (qPCR) assay for bigheaded carp eDNA was one to two orders of magnitude more sensitive than the existing endpoint PCR assays. When applied to eDNA samples from an experimental pond containing bigheaded carp, the qPCR assay produced a detection probability of 94.8% compared to 4.2% for the endpoint PCR assays. Also, the eDNA capture and extraction method we adapted from aquatic microbiology yielded five times more bigheaded carp eDNA from the experimental pond than the standard method, at a per sample cost over forty times lower. Our new, more sensitive assay provides a quantitative tool for eDNA-based monitoring of bigheaded carp, and the higher-yielding eDNA capture and extraction method we describe can be used for eDNA-based monitoring of any aquatic species. PMID:25474207

  1. Improved methods for capture, extraction, and quantitative assay of environmental DNA from Asian bigheaded carp (Hypophthalmichthys spp.).

    PubMed

    Turner, Cameron R; Miller, Derryl J; Coyne, Kathryn J; Corush, Joel

    2014-01-01

    Indirect, non-invasive detection of rare aquatic macrofauna using aqueous environmental DNA (eDNA) is a relatively new approach to population and biodiversity monitoring. As such, the sensitivity of monitoring results to different methods of eDNA capture, extraction, and detection is being investigated in many ecosystems and species. One of the first and largest conservation programs with eDNA-based monitoring as a central instrument focuses on Asian bigheaded carp (Hypophthalmichthys spp.), an invasive fish spreading toward the Laurentian Great Lakes. However, the standard eDNA methods of this program have not advanced since their development in 2010. We developed new, quantitative, and more cost-effective methods and tested them against the standard protocols. In laboratory testing, our new quantitative PCR (qPCR) assay for bigheaded carp eDNA was one to two orders of magnitude more sensitive than the existing endpoint PCR assays. When applied to eDNA samples from an experimental pond containing bigheaded carp, the qPCR assay produced a detection probability of 94.8% compared to 4.2% for the endpoint PCR assays. Also, the eDNA capture and extraction method we adapted from aquatic microbiology yielded five times more bigheaded carp eDNA from the experimental pond than the standard method, at a per sample cost over forty times lower. Our new, more sensitive assay provides a quantitative tool for eDNA-based monitoring of bigheaded carp, and the higher-yielding eDNA capture and extraction method we describe can be used for eDNA-based monitoring of any aquatic species.

  2. 40 CFR 258.54 - Detection monitoring program.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... FOR MUNICIPAL SOLID WASTE LANDFILLS Ground-Water Monitoring and Corrective Action § 258.54 Detection... reaction products in the unsaturated zone beneath the MSWLF unit; (iii) The detectability of indicator parameters, waste constituents, and reaction products in the ground water; and (iv) The concentration or...

  3. A review on technologies and their usage in solid waste monitoring and management systems: Issues and challenges

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hannan, M.A., E-mail: hannan@eng.ukm.my; Abdulla Al Mamun, Md., E-mail: md.abdulla@siswa.ukm.edu.my; Hussain, Aini, E-mail: aini@eng.ukm.my

    Highlights: • Classification of available technologies for SWM system in four core category. • Organization of technology based SWM systems in three main groups. • Summary of SWM systems with target application, methodology and functional domain. • Issues and challenges are highlighted for further design of a sustainable system. - Abstract: In the backdrop of prompt advancement, information and communication technology (ICT) has become an inevitable part to plan and design of modern solid waste management (SWM) systems. This study presents a critical review of the existing ICTs and their usage in SWM systems to unfold the issues and challengesmore » towards using integrated technologies based system. To plan, monitor, collect and manage solid waste, the ICTs are divided into four categories such as spatial technologies, identification technologies, data acquisition technologies and data communication technologies. The ICT based SWM systems classified in this paper are based on the first three technologies while the forth one is employed by almost every systems. This review may guide the reader about the basics of available ICTs and their application in SWM to facilitate the search for planning and design of a sustainable new system.« less

  4. Final report on the waste area grouping perimeter groundwater quality monitoring well installation program at Oak Ridge National Laboratory, Oak Ridge, Tennessee

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Greene, J.A.

    1991-06-01

    A groundwater quality monitoring well installation program was conducted at Oak Ridge National Laboratory (ORNL) to meet the requirements of environmental regulations, including the Resource Conservation and Recovery Act (RCRA) and the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA). A total of 173 wells were installed and developed at 11 different waste area groupings (WAGs) between June 1986 and November 1990. A location map of the wells is included.

  5. Thermal and enzymatic recovering of proteins from untanned leather waste.

    PubMed

    Bajza, Z; Vrucek, V

    2001-01-01

    The laboratory trials of a process to treat untanned leather waste to isolate valuable protein products are presented. In this comparative study, both thermal and enzymatic treatments of leather waste were performed. The enzymatic method utilizes commercially available alkaline protease at moderate temperatures and for short periods of time. The concentration of the enzyme was 500 units per gram of leather waste which makes the method cost-effective. Amino acid composition in the hydrolysate obtained by the enzyme hydrolysis of untanned leather waste is determined. Chemical and physical properties of protein powder products from untanned leather waste were evaluated by spectrophotometric and chromatographic methods and by use of electron microscope. The results of microbiological assays confirm that these products agree to food safety standards. This relatively simple treatment of untanned leather waste may provide a practical and economical solution to the disposal of potentially dangerous waste.

  6. Quantitative CrAssphage PCR Assays for Human Fecal Pollution Measurement

    EPA Science Inventory

    Environmental waters are monitored for fecal pollution to protect public health and water resources. Traditionally, general fecal indicator bacteria are used; however, they cannot distinguish human fecal waste from pollution from other animals. Recently, a novel bacteriophage, cr...

  7. Immunologic monitoring of cancer vaccine therapy: results of a workshop sponsored by the Society for Biological Therapy.

    PubMed

    Keilholz, Ulrich; Weber, Jeffrey; Finke, James H; Gabrilovich, Dmitry I; Kast, W Martin; Disis, Mary L; Kirkwood, John M; Scheibenbogen, Carmen; Schlom, Jeff; Maino, Vernon C; Lyerly, H Kim; Lee, Peter P; Storkus, Walter; Marincola, Franceso; Worobec, Alexandra; Atkins, Michael B

    2002-01-01

    The Society for Biological Therapy held a Workshop last fall devoted to immune monitoring for cancer immunotherapy trials. Participants included members of the academic and pharmaceutical communities as well as the National Cancer Institute and the Food and Drug Administration. Discussion focused on the relative merits and appropriate use of various immune monitoring tools. Six breakout groups dealt with assays of T-cell function, serologic and proliferation assays to assess B cell and T helper cell activity, and enzyme-linked immunospot assay, tetramer, cytokine flow cytometry, and reverse transcription polymerase chain reaction assays of T-cell immunity. General conclusions included: (1) future vaccine studies should be designed to determine whether T-cell dysfunction (tumor-specific and nonspecific) correlated with clinical outcome; (2) tetramer-based assays yield quantitative but not functional data (3) enzyme-linked immunospot assays have the lowest limit of detection (4) cytokine flow cytometry have a higher limit of detection than enzyme-linked immunospot assay, but offer the advantages of speed and the ability to identify subsets of reactive cells; (5) antibody tests are simple and accurate and should be incorporated to a greater extent in monitoring plans; (6) proliferation assays are imprecise and should not be emphasized in future studies; (7) the reverse transcription polymerase chain reaction assay is a promising research approach that is not ready for widespread application; and (8)there is a critical need to validate these assays as surrogates for vaccine potency and clinical effect. Current data and opinion support the use of a functional assay like the enzyme-linked immunospot assay or cytokine flow cytometry in combination with a quantitative assay like tetramers for immune monitoring. At present, assays appear to be most useful as measures of vaccine potency. Careful immune monitoring in association with larger scale clinical trials ultimately

  8. Monitoring and Inventory of Hazardous Pollutants Emissions from Solid Waste Open Burning

    NASA Astrophysics Data System (ADS)

    KIM Oanh, N. T.

    2017-12-01

    Open burning remains a way to dispose of solid waste in developing countries, commonly practiced in places where municipal solid waste (MSW) management is not sufficiently efficient. This open burning practice emits huge amounts of toxic air pollutants, including semi-VOC of dioxins (PCDD/F) and polycyclic aromatic hydrocarbons (PAHs), particulate matters (PM), and toxic gases. This study measured toxic substances released from simulated open burning experiments of MSW (5 batches) and plastic waste (3 batches) to determine emission factors (EFs). Carbon content of the waste before and after burning was measured and the EFs were calculated using the carbon balance method. The obtained EFs of CO; CO2; NO2 and SO2 were 102±61; 1,684±96; 0.69±0.54; and 1.44±1.18 g/kg of dry MSW. The corresponding values for plastic waste were 13.0±6.0; 1,824±10; 0.12±0.07; and 0.14±0.09 g/kg, respectively. The EF of coarse PM (PM10-2.5); PM2.5 and BC in PM2.5 were, respectively, 6.1±5.6; 6.4±5.1 and 1.1±0.7 g/kg of MSW, and 2.3±0.1; 2.5±0.3; and 0.2±0.2 g/kg of plastic waste. The EFs of 17 dioxins and 16 PAHs were respectively 1,050±500 ng-TEQ dioxins (70% in gas phase) and 117± 21 mg PAHs (92% in gas phase) per kg of MSW, while those for plastic waste were 8.6 ng TEQ dioxins (36% in gas phase) and 85.2±0.06 mg PAHs (99% in gas phase). Using the activity data from own surveys and literature, we estimated the annual emissions from solid waste open burning in Southeast Asia for 2015. Higher emissions in the domain were seen during the dry months and over large urban areas. The large amounts of toxic pollutants emitted from this open burning activity call for actions to stop this practice which in turn requires integrated environmental management approach simultaneously considering both solid waste and air pollution.

  9. BP Spill Sampling and Monitoring Data

    EPA Pesticide Factsheets

    This dataset analyzes waste from the the British Petroleum Deepwater Horizon Rig Explosion Emergency Response, providing opportunity to query data sets by metadata criteria and find resulting raw datasets in CSV format.The data query tool allows users to download EPA's air, water and sediment sampling and monitoring data that has been collected in response to the BP oil spill. All sampling and monitoring data that has been collected to date is available for download as raw structured data.The query tools enables CSV file creation to be refined based on the following search criteria: date range (between April 28, 2010 and 9/29/2010); location by zip, city, or county; media (solid waste, weathered oil, air, surface water, liquid waste, tar, sediment, water); substance categories (based on media selection) and substances (based on substance category selection).

  10. 40 CFR 271.12 - Requirements for hazardous waste management facilities.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... and 266. These standards shall include: (a) Technical standards for tanks, containers, waste piles...-closure monitoring and maintenance; (e) Groundwater monitoring; (f) Security to prevent unauthorized access to the facility; (g) Facility personnel training; (h) Inspections, monitoring, recordkeeping, and...

  11. 40 CFR 271.12 - Requirements for hazardous waste management facilities.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... and 266. These standards shall include: (a) Technical standards for tanks, containers, waste piles...-closure monitoring and maintenance; (e) Groundwater monitoring; (f) Security to prevent unauthorized access to the facility; (g) Facility personnel training; (h) Inspections, monitoring, recordkeeping, and...

  12. 40 CFR 271.12 - Requirements for hazardous waste management facilities.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... and 266. These standards shall include: (a) Technical standards for tanks, containers, waste piles...-closure monitoring and maintenance; (e) Groundwater monitoring; (f) Security to prevent unauthorized access to the facility; (g) Facility personnel training; (h) Inspections, monitoring, recordkeeping, and...

  13. 40 CFR 271.12 - Requirements for hazardous waste management facilities.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... and 266. These standards shall include: (a) Technical standards for tanks, containers, waste piles...-closure monitoring and maintenance; (e) Groundwater monitoring; (f) Security to prevent unauthorized access to the facility; (g) Facility personnel training; (h) Inspections, monitoring, recordkeeping, and...

  14. 40 CFR 271.12 - Requirements for hazardous waste management facilities.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... and 266. These standards shall include: (a) Technical standards for tanks, containers, waste piles...-closure monitoring and maintenance; (e) Groundwater monitoring; (f) Security to prevent unauthorized access to the facility; (g) Facility personnel training; (h) Inspections, monitoring, recordkeeping, and...

  15. Comparison of the Second-Generation Digene Hybrid Capture Assay with the Branched-DNA Assay for Measurement of Hepatitis B Virus DNA in Serum

    PubMed Central

    Ho, Stephen K. N.; Chan, Tak Mao; Cheng, Ignatius K. P.; Lai, Kar Neng

    1999-01-01

    The optimal hepatitis B virus (HBV) DNA quantitative assay for clinical use remains to be determined. We examined the sensitivity, linearity, and variability of a novel second-generation antibody capture solution hybridization assay, the Digene Hybrid Capture II assay (HCII), and compared it with another widely used solution hybridization assay, the branched-DNA (bDNA) assay (Quantiplex; Chiron Corp.). Our results showed similar and satisfactory assay linearity values, as well as interassay and intra-assay variability values, for both HCII and bDNA assays across different ranges of HBV DNA. Ninety-one percent of 102 serum samples from hepatitis B surface antigen-positive patients showed concordant results with the two assays. The HCII assay was more sensitive than the bDNA assay by 1 dilution, with the lowest reading being 0.9 pg/ml (3.8 pg/ml by bDNA assay). The HBV DNA seropositivity rates for the 102 samples were 58, 67, and 97% by bDNA, HCII, and nested PCR, respectively. While the relationship between results obtained with the bDNA assay and those with the HCII assay was nonlinear, with the bDNA assay yielding values 2.83 ± 0.92-fold higher than those of the HCII assay, especially at high HBV DNA levels, a linear relationship was observed between the two sets of data after logarithmic conversion. The formula for interassay conversion of results was derived as follows: HBV DNA by HCII (picograms per milliliter) = 3.19 × [HBV DNA by bDNA (megaequivalents per milliliter)]0.866. The HCII assay was technically less complex and required a shorter assay time (4 h) than the bDNA assay (24 h). We conclude that the HCII assay compares favorably with the bDNA assay and offers the additional advantages of increased sensitivity and shorter assay time. The increased sensitivity should be particularly useful in monitoring the efficacy of antiviral therapies and detecting the emergence of drug-resistant HBV mutants. PMID:10405385

  16. Comparison of the second-generation digene hybrid capture assay with the branched-DNA assay for measurement of hepatitis B virus DNA in serum.

    PubMed

    Ho, S K; Chan, T M; Cheng, I K; Lai, K N

    1999-08-01

    The optimal hepatitis B virus (HBV) DNA quantitative assay for clinical use remains to be determined. We examined the sensitivity, linearity, and variability of a novel second-generation antibody capture solution hybridization assay, the Digene Hybrid Capture II assay (HCII), and compared it with another widely used solution hybridization assay, the branched-DNA (bDNA) assay (Quantiplex; Chiron Corp.). Our results showed similar and satisfactory assay linearity values, as well as interassay and intra-assay variability values, for both HCII and bDNA assays across different ranges of HBV DNA. Ninety-one percent of 102 serum samples from hepatitis B surface antigen-positive patients showed concordant results with the two assays. The HCII assay was more sensitive than the bDNA assay by 1 dilution, with the lowest reading being 0.9 pg/ml (3.8 pg/ml by bDNA assay). The HBV DNA seropositivity rates for the 102 samples were 58, 67, and 97% by bDNA, HCII, and nested PCR, respectively. While the relationship between results obtained with the bDNA assay and those with the HCII assay was nonlinear, with the bDNA assay yielding values 2.83 +/- 0.92-fold higher than those of the HCII assay, especially at high HBV DNA levels, a linear relationship was observed between the two sets of data after logarithmic conversion. The formula for interassay conversion of results was derived as follows: HBV DNA by HCII (picograms per milliliter) = 3.19 x [HBV DNA by bDNA (megaequivalents per milliliter)](0.866). The HCII assay was technically less complex and required a shorter assay time (4 h) than the bDNA assay (24 h). We conclude that the HCII assay compares favorably with the bDNA assay and offers the additional advantages of increased sensitivity and shorter assay time. The increased sensitivity should be particularly useful in monitoring the efficacy of antiviral therapies and detecting the emergence of drug-resistant HBV mutants.

  17. An embedded system developed for hand held assay used in water monitoring

    NASA Astrophysics Data System (ADS)

    Wu, Lin; Wang, Jianwei; Ramakrishna, Bharath; Hsueh, Mingkai; Liu, Jonathan; Wu, Qufei; Wu, Chao-Cheng; Cao, Mang; Chang, Chein-I.; Jensen, Janet L.; Jensen, James O.; Knapp, Harlan; Daniel, Robert; Yin, Ray

    2005-11-01

    The US Army Joint Service Agent Water Monitor (JSAWM) program is currently interested in an approach that can implement a hardware- designed device in ticket-based hand-held assay (currently being developed) used for chemical/biological agent detection. This paper presents a preliminary investigation of the proof of concept. Three components are envisioned to accomplish the task. One is the ticket development which has been undertaken by the ANP, Inc. Another component is the software development which has been carried out by the Remote Sensing Signal and Image Processing Laboratory (RSSIPL) at the University of Maryland, Baltimore County (UMBC). A third component is an embedded system development which can be used to drive the UMBC-developed software to analyze the ANP-developed HHA tickets on a small pocket-size device like a PDA. The main focus of this paper is to investigate the third component that is viable and is yet to be explored. In order to facilitate to prove the concept, a flatbed scanner is used to replace a ticket reader to serve as an input device. The Stargate processor board is used as the embedded System with Embedded Linux installed. It is connected to an input device such as scanner as well as output devices such as LCD display or laptop etc. It executes the C-Coded processing program developed for this embedded system and outputs its findings on a display device. The embedded system to be developed and investigated in this paper is the core of a future hardware device. Several issues arising in such an embedded system will be addressed. Finally, the proof-of-concept pilot embedded system will be demonstrated.

  18. [Use of comet assay for the risk assessment of oil- and chemical-industry workers].

    PubMed

    Megyesi, János; Biró, Anna; Wigmond, László; Major, Jenő; Tompa, Anna

    2014-11-23

    The comet assay is a fluorescent microscopic method that is able to detect DNA strand-breaks even in non-proliferative cells in samples with low cell counts. The aim of the authors was to measure genotoxic DNA damage and assess oxidative DNA damage caused by occupational exposure in groups exposed to benzene, polycyclic aromatic carbohydrates and styrene at the workplace in order to clarify whether the comet assay can be used as an effect marker tool in genotoxicology monitoring. In addition to the basic steps of the comet assay, one sample was treated with formamido-pirimidine-DNA-glycolase restriction-enzyme that measures oxidative DNA damage. An increase was observed in tail moments in each group of untreated and Fpg-treated samples compared to the control. It can be concluded that occupational exposure can be detected with the method. The comet assay may prove to be an excellent effect marker and a supplementary technique for monitoring the presence or absence of genotoxic effects.

  19. Particle Swarm Imaging (PSIM) - Innovative Gamma-Ray Assay - 13497

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Parvin, Daniel; Clarke, Sean; Humes, Sarah J.

    2013-07-01

    Particle Swarm Imaging is an innovative technique used to perform quantitative gamma-ray assay. The innovation overcomes some of the difficulties associated with the accurate measurement and declaration of measurement uncertainties of radionuclide inventories within waste items when the distribution of activity is unknown. Implementation requires minimal equipment, with field measurements and results obtained using only a single electrically cooled HRGS gamma-ray detector. Examples of its application in the field are given in this paper. (authors)

  20. ELISPOT Assays in 384-Well Format: Up to 30 Data Points with One Million Cells

    PubMed Central

    Hanson, Jodi; Sundararaman, Srividya; Caspell, Richard; Karacsony, Edith; Karulin, Alexey Y.; Lehmann, Paul V.

    2015-01-01

    Comprehensive immune monitoring requires that frequencies of T cells, producing different cytokines, are measured to establish the magnitude of Th1, Th2, and Th17 components of cell-mediated immunity. Antigen titration provides additional information about the affinity of T cell response. In tumor immunity, it is also advisable to account for determinant spreading by testing multiple epitopes. Efforts for comprehensive immune monitoring would require substantial numbers of PBMC to run the above tests systematically, which in most test cases is limiting. Immune monitoring with ELISPOT assays have been performed, thus far, in a 96-well format. In this study we show that one can increase cell utilization by performing the assay in 384-well plates whose membrane surface area is one third that of 96-well plates. Systematic testing of PBMC for antigen-specific T cell response in the two formats demonstrated that the 384-well assay corresponds to a one-in-three miniaturization of the 96-well assay. The lowest number of cells that can be used in the 384-well format, while allowing for sufficient contact with APC, is 33,000 PBMC/well. Therefore, with one million PBMC typically obtained from 1 mL of blood, a 30 well T cell ELISPOT assay can be performed in a 384-well format. PMID:25643292

  1. 33 CFR 151.1516 - Compliance monitoring.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 2 2011-07-01 2011-07-01 false Compliance monitoring. 151.1516...) POLLUTION VESSELS CARRYING OIL, NOXIOUS LIQUID SUBSTANCES, GARBAGE, MUNICIPAL OR COMMERCIAL WASTE, AND... River § 151.1516 Compliance monitoring. (a) The master of each vessel equipped with ballast tanks shall...

  2. Geodiametris: an integrated geoinformatic approach for monitoring land pollution from the disposal of olive oil mill wastes

    NASA Astrophysics Data System (ADS)

    Alexakis, Dimitrios D.; Sarris, Apostolos; Papadopoulos, Nikos; Soupios, Pantelis; Doula, Maria; Cavvadias, Victor

    2014-08-01

    The olive-oil industry is one of the most important sectors of agricultural production in Greece, which is the third in olive-oil production country worldwide. Olive oil mill wastes (OOMW) constitute a major factor in pollution in olivegrowing regions and an important problem to be solved for the agricultural industry. The olive-oil mill wastes are normally deposited at tanks, or directly in the soil or even on adjacent torrents, rivers and lakes posing a high risk to the environmental pollution and the community health. GEODIAMETRIS project aspires to develop integrated geoinformatic methodologies for performing monitoring of land pollution from the disposal of OOMW in the island of Crete -Greece. These methodologies integrate GPS surveys, satellite remote sensing and risk assessment analysis in GIS environment, application of in situ and laboratory geophysical methodologies as well as soil and water physicochemical analysis. Concerning project's preliminary results, all the operating OOMW areas located in Crete have been already registered through extensive GPS field campaigns. Their spatial and attribute information has been stored in an integrated GIS database and an overall OOMW spectral signature database has been constructed through the analysis of multi-temporal Landsat-8 OLI satellite images. In addition, a specific OOMW area located in Alikianos village (Chania-Crete) has been selected as one of the main case study areas. Various geophysical methodologies, such as Electrical Resistivity Tomography, Induced Polarization, multifrequency electromagnetic, Self Potential measurements and Ground Penetrating Radar have been already implemented. Soil as well as liquid samples have been collected for performing physico-chemical analysis. The preliminary results have already contributed to the gradual development of an integrated environmental monitoring tool for studying and understanding environmental degradation from the disposal of OOMW.

  3. Performance test results of noninvasive characterization of Resource Conservation and Recovery Act surrogate waste by prompt gamma neutron activation analysis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gehrke, R.J.; Streier, G.G.

    1997-03-01

    During FY-96, a performance test was carried out with funding from the Mixed Waste Focus Area (MWFA) of the Department of Energy (DOE) to determine the noninvasive elemental assay capabilities of commercial companies for Resource Conservation and Recovery Act (RCRA) metals present in 8-gal drums containing surrogate waste. Commercial companies were required to be experienced in the use of prompt gamma neutron activation analysis (PGNAA) techniques and to have a prototype assay system with which to conduct the test assays. Potential participants were identified through responses to a call for proposals advertised in the Commerce Business Daily and through personalmore » contacts. Six companies were originally identified. Two of these six were willing and able to participate in the performance test, as described in the test plan, with some subsidizing from the DOE MWFA. The tests were conducted with surrogate sludge waste because (1) a large volume of this type of waste awaits final disposition and (2) sludge tends to be somewhat homogeneous. The surrogate concentrations of the above RCRA metals ranged from {approximately} 300 ppm to {approximately} 20,000 ppm. The lower limit was chosen as an estimate of the expected sensitivity of detection required by noninvasive, pretreatment elemental assay systems to be of value for operational and compliance purposes and to still be achievable with state-of-the-art methods of analysis. The upper limit of {approximately} 20,000 ppm was chosen because it is the opinion of the author that assay above this concentration level is within current state-of-the-art methods for most RCRA constituents. This report is organized into three parts: Part 1, Test Plan to Evaluate the Technical Status of Noninvasive Elemental Assay Techniques for Hazardous Waste; Part 2, Participants` Results; and Part 3, Evaluation of and Comments on Participants` Results.« less

  4. Detection of Streptococcus pyogenes using rapid visual molecular assay.

    PubMed

    Zhao, Xiangna; He, Xiaoming; Li, Huan; Zhao, Jiangtao; Huang, Simo; Liu, Wei; Wei, Xiao; Ding, Yiwei; Wang, Zhaoyan; Zou, Dayang; Wang, Xuesong; Dong, Derong; Yang, Zhan; Yan, Xiabei; Huang, Liuyu; Du, Shuangkui; Yuan, Jing

    2015-09-01

    Streptococcus pyogenes is an increasingly important pathogen in many parts of the world. Rapid and accurate detection of S. pyogenes aids in the control of the infection. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed and validated for the specific detection of S. pyogenes. The assay incorporates two methods: a chromogenic analysis using a calcein/Mn(2+) complex and real-time turbidity monitoring to assess the reaction. Both methods detected the target DNA within 60 min under 64°C isothermal conditions. The assay used specifically designed primers to target spy1258, and correctly identified 111 strains of S. pyogenes and 32 non-S. pyogenes strains, including other species of the genus Streptococcus. Tests using reference strains showed that the LAMP assay was highly specific. The sensitivity of the assay, with a detection limit of 1.49 pg DNA, was 10-fold greater than that of PCR. The LAMP assay established in this study is simple, fast and sensitive, and does not rely upon any special equipment; thus, it could be employed in clinical diagnosis. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  5. 2011 Annual Industrial Wastewater Reuse Report for the Idaho National Laboratory Site's Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    David Frederick

    2012-02-01

    This report describes conditions, as required by the state of Idaho Industrial Wastewater Reuse Permit (LA-000160-01), for the wastewater reuse site at the Idaho National Laboratory Site's Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond from November 1, 2010 through October 31, 2011. The report contains the following information: (1) Facility and system description; (2) Permit required effluent monitoring data and loading rates; (3) Groundwater monitoring data; (4) Status of special compliance conditions; and (5) Discussion of the facility's environmental impacts. During the 2011 reporting year, an estimated 6.99 million gallons of wastewater were discharged to themore » Industrial Waste Ditch and Pond which is well below the permit limit of 13 million gallons per year. Using the dissolved iron data, the concentrations of all permit-required analytes in the samples from the down gradient monitoring wells were below the Ground Water Quality Rule Primary and Secondary Constituent Standards.« less

  6. 2010 Annual Industrial Wastewater Reuse Report for the Idaho National Laboratory Site's Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    David B. Frederick

    2011-02-01

    This report describes conditions, as required by the state of Idaho Industrial Wastewater Reuse Permit (#LA 000160 01), for the wastewater reuse site at the Idaho National Laboratory Site’s Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond from May 1, 2010 through October 31, 2010. The report contains the following information: • Facility and system description • Permit required effluent monitoring data and loading rates • Groundwater monitoring data • Status of special compliance conditions • Discussion of the facility’s environmental impacts During the 2010 partial reporting year, an estimated 3.646 million gallons of wastewater were dischargedmore » to the Industrial Waste Ditch and Pond which is well below the permit limit of 13 million gallons per year. The concentrations of all permit-required analytes in the samples from the down gradient monitoring wells were below the Ground Water Quality Rule Primary and Secondary Constituent Standards.« less

  7. Differential sensitivity of von Willebrand factor (VWF) 'activity' assays to large and small VWF molecular weight forms: a cross-laboratory study comparing ristocetin cofactor, collagen-binding and mAb-based assays.

    PubMed

    Favaloro, E J; Bonar, R; Chapman, K; Meiring, M; Funk Adcock, D

    2012-06-01

    von Willebrand disease (VWD), the most common inherited bleeding disorder, is caused by deficiencies and/or defects in von Willebrand factor (VWF). An effective diagnostic and VWD typing strategy requires plasma testing for factor VIII, and VWF antigen plus one or more VWF 'activity' assays. VWF activity is classically assessed by using VWF ristocetin cofactor activity (VWF:RCo), although VWF collagen-binding (VWF:CB) and VWF mAb-based (VWF activity [VWF:Act]) assays are used by some laboratories. To perform a cross-laboratory study to specifically evaluate these three VWF activity assays for comparative sensitivity to loss of high molecular weight (HMW) VWF, representing the form of VWF that is most functionally active and that is absent in some types of VWD, namely 2A and 2B. A set of eight samples, including six selectively representing stepwise reduction in HMW VWF, were tested by 51 different laboratories using a variety of assays. The combined data showed that the VWF:CB and VWF:RCo assays had higher sensitivity to the loss of HMW VWF than did the VWF:Act assay. Moreover, within-method analysis identified better HMW VWF sensitivity of some VWF:CB assays than of others, with all VWF:CB assays still showing better sensitivity than the VWF:Act assay. Differences were also identified between VWF:RCo methodologies on the basis of either platelet aggregometry or as performed on automated analyzers. We believe that these results have significant clinical implications for the diagnosis of VWD and monitoring of its therapy, as well as for the future diagnosis and therapy monitoring of thrombotic thrombocytopenic purpura. © 2012 International Society on Thrombosis and Haemostasis.

  8. Modification of an environmental surveillance program to monitor PCDD/Fs and metals around a municipal solid waste incinerator.

    PubMed

    Vilavert, Lolita; Nadal, Martí; Mari, Montse; Schuhmacher, Marta; Domingo, José L

    2009-11-01

    Since the mid-90s, an environmental surveillance program has been on-going to provide information on the levels of PCDD/Fs and various metals in soil and vegetation samples collected in the vicinity of a municipal solid waste incinerator (MSWI) in Tarragona (Catalonia, Spain). However, the presence of other potential sources of pollution in the zone, such as traffic, forest fires, local industries, etc., makes hard to determine the impact concerning the MSWI. Therefore, in 2007 a change in the monitoring program was implemented by collecting additional ambient air samples through active and passive sampling devices. Mean PCDD/F levels in herbage and soil were 0.10 ng I-TEQ/kg dry weight (range: 0.05-0.17 ng I-TEQ/kg dw) and 0.64 ng I-TEQ/kg dw (range: 0.13-2.41 ng I-TEQ/kg dw), respectively. A significant reduction of the PCDD/F concentration in both monitors was observed with respect to our previous surveys. Air mean concentrations of PCDD/Fs were 12.04 and 15.21 fg WHO-TEQ/m(3) in 2007 and 2008, respectively, meaning a non-significant increase of 26%. In addition, a generalized increase of environmental metal levels with respect to our baseline study was not observed. The current concentrations of PCDD/Fs and metals in the vicinity of the MSWI of Tarragona are relatively low in comparison with other areas under the influence of emissions from waste incinerators. This indicates that the environmental impact of the MSWI of Tarragona is not significant. Moreover, the modification of the surveillance program has proven to be successful.

  9. Characterization monitoring & sensor technology crosscutting program

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    NONE

    1996-08-01

    The purpose of the Characterization, Monitoring, and Sensor Technology Crosscutting Program (CMST-CP) is to deliver appropriate characterization, monitoring, and sensor technology (CMST) to the OFfice of Waste Management (EM-30), the Office of Environmental Restoration (EM-40), and the Office of Facility Transition and Management (EM-60).

  10. 40 CFR 437.4 - Monitoring requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    .... (a) Permit compliance monitoring is required for each regulated parameter. (b) Any CWT facility that... subpart. (3) When a CWT facility treats any waste receipt that contains cyanide at a concentration higher than 136 mg/L, the CWT facility must monitor for cyanide after cyanide treatment and before dilution...

  11. 40 CFR 437.4 - Monitoring requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    .... (a) Permit compliance monitoring is required for each regulated parameter. (b) Any CWT facility that... subpart. (3) When a CWT facility treats any waste receipt that contains cyanide at a concentration higher than 136 mg/L, the CWT facility must monitor for cyanide after cyanide treatment and before dilution...

  12. 40 CFR 437.4 - Monitoring requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    .... (a) Permit compliance monitoring is required for each regulated parameter. (b) Any CWT facility that... subpart. (3) When a CWT facility treats any waste receipt that contains cyanide at a concentration higher than 136 mg/L, the CWT facility must monitor for cyanide after cyanide treatment and before dilution...

  13. Recombinant antigen-based antibody assays for the diagnosis and surveillance of lymphatic filariasis – a multicenter trial

    PubMed Central

    Lammie, Patrick J; Weil, Gary; Noordin, Rahmah; Kaliraj, Perumal; Steel, Cathy; Goodman, David; Lakshmikanthan, Vijaya B; Ottesen, Eric

    2004-01-01

    The development of antifilarial antibody responses is a characteristic feature of infection with filarial parasites. It should be possible to exploit this fact to develop tools to monitor the progress of the global program to eliminate lymphatic filariasis (LF); however, assays based on parasite extracts suffer from a number of limitations, including the paucity of parasite material, the difficulty of assay standardization and problems with assay specificity. In principle, assays based on recombinant filarial antigens should address these limitations and provide useful tools for diagnosis and surveillance of LF. The present multicenter study was designed to compare the performance of antibody assays for filariasis based on recombinant antigens Bm14, WbSXP, and BmR1. Coded serum specimens were distributed to five participating laboratories where assays for each antigen were conducted in parallel. Assays based on Bm14, WbSXP, or BmR1 demonstrated good sensitivity (>90%) for field use and none of the assays demonstrated reactivity with specimens from persons with non-filarial helminth infections. Limitations of the assays are discussed. Well-designed field studies are now needed to assess sampling methodology and the application of antibody testing to the monitoring and surveillance of LF elimination programs. PMID:15347425

  14. 40 CFR 258.54 - Detection monitoring program.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... FOR MUNICIPAL SOLID WASTE LANDFILLS Ground-Water Monitoring and Corrective Action § 258.54 Detection... indicator parameters for a MSWLF unit, in lieu of some or all of the heavy metals (constituents 1-15 in... parameters, waste constituents, and reaction products in the ground water; and (iv) The concentration or...

  15. 40 CFR 258.54 - Detection monitoring program.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... FOR MUNICIPAL SOLID WASTE LANDFILLS Ground-Water Monitoring and Corrective Action § 258.54 Detection... indicator parameters for a MSWLF unit, in lieu of some or all of the heavy metals (constituents 1-15 in... parameters, waste constituents, and reaction products in the ground water; and (iv) The concentration or...

  16. 40 CFR 258.54 - Detection monitoring program.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... FOR MUNICIPAL SOLID WASTE LANDFILLS Ground-Water Monitoring and Corrective Action § 258.54 Detection... indicator parameters for a MSWLF unit, in lieu of some or all of the heavy metals (constituents 1-15 in... parameters, waste constituents, and reaction products in the ground water; and (iv) The concentration or...

  17. 40 CFR 258.54 - Detection monitoring program.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... FOR MUNICIPAL SOLID WASTE LANDFILLS Ground-Water Monitoring and Corrective Action § 258.54 Detection... indicator parameters for a MSWLF unit, in lieu of some or all of the heavy metals (constituents 1-15 in... parameters, waste constituents, and reaction products in the ground water; and (iv) The concentration or...

  18. Standardizing a simpler, more sensitive and accurate tail bleeding assay in mice

    PubMed Central

    Liu, Yang; Jennings, Nicole L; Dart, Anthony M; Du, Xiao-Jun

    2012-01-01

    AIM: To optimize the experimental protocols for a simple, sensitive and accurate bleeding assay. METHODS: Bleeding assay was performed in mice by tail tip amputation, immersing the tail in saline at 37 °C, continuously monitoring bleeding patterns and measuring bleeding volume from changes in the body weight. Sensitivity and extent of variation of bleeding time and bleeding volume were compared in mice treated with the P2Y receptor inhibitor prasugrel at various doses or in mice deficient of FcRγ, a signaling protein of the glycoprotein VI receptor. RESULTS: We described details of the bleeding assay with the aim of standardizing this commonly used assay. The bleeding assay detailed here was simple to operate and permitted continuous monitoring of bleeding pattern and detection of re-bleeding. We also reported a simple and accurate way of quantifying bleeding volume from changes in the body weight, which correlated well with chemical assay of hemoglobin levels (r2 = 0.990, P < 0.0001). We determined by tail bleeding assay the dose-effect relation of the anti-platelet drug prasugrel from 0.015 to 5 mg/kg. Our results showed that the correlation of bleeding time and volume was unsatisfactory and that compared with the bleeding time, bleeding volume was more sensitive in detecting a partial inhibition of platelet’s haemostatic activity (P < 0.01). Similarly, in mice with genetic disruption of FcRγ as a signaling molecule of P-selectin glycoprotein ligand-1 leading to platelet dysfunction, both increased bleeding volume and repeated bleeding pattern defined the phenotype of the knockout mice better than that of a prolonged bleeding time. CONCLUSION: Determination of bleeding pattern and bleeding volume, in addition to bleeding time, improved the sensitivity and accuracy of this assay, particularly when platelet function is partially inhibited. PMID:24520531

  19. Potential for Gulls to Transport Bacteria from Human Waste Sites to Beaches

    EPA Science Inventory

    Contamination of recreational beaches due to fecal waste from gulls complicates beach monitoring and may pose a risk to public health. Gulls that feed at human waste sites may ingest human-associated fecal microorganisms associated with that waste. If these gulls also visit beach...

  20. Rapid automation of a cell-based assay using a modular approach: case study of a flow-based Varicella Zoster Virus infectivity assay.

    PubMed

    Joelsson, Daniel; Gates, Irina V; Pacchione, Diana; Wang, Christopher J; Bennett, Philip S; Zhang, Yuhua; McMackin, Jennifer; Frey, Tina; Brodbeck, Kristin C; Baxter, Heather; Barmat, Scott L; Benetti, Luca; Bodmer, Jean-Luc

    2010-06-01

    Vaccine manufacturing requires constant analytical monitoring to ensure reliable quality and a consistent safety profile of the final product. Concentration and bioactivity of active components of the vaccine are key attributes routinely evaluated throughout the manufacturing cycle and for product release and dosage. In the case of live attenuated virus vaccines, bioactivity is traditionally measured in vitro by infection of susceptible cells with the vaccine followed by quantification of virus replication, cytopathology or expression of viral markers. These assays are typically multi-day procedures that require trained technicians and constant attention. Considering the need for high volumes of testing, automation and streamlining of these assays is highly desirable. In this study, the automation and streamlining of a complex infectivity assay for Varicella Zoster Virus (VZV) containing test articles is presented. The automation procedure was completed using existing liquid handling infrastructure in a modular fashion, limiting custom-designed elements to a minimum to facilitate transposition. In addition, cellular senescence data provided an optimal population doubling range for long term, reliable assay operation at high throughput. The results presented in this study demonstrate a successful automation paradigm resulting in an eightfold increase in throughput while maintaining assay performance characteristics comparable to the original assay. Copyright 2010 Elsevier B.V. All rights reserved.

  1. Composting of food wastes: Status and challenges.

    PubMed

    Cerda, Alejandra; Artola, Adriana; Font, Xavier; Barrena, Raquel; Gea, Teresa; Sánchez, Antoni

    2018-01-01

    This review analyses the main challenges of the process of food waste composting and examines the crucial aspects related to the quality of the produced compost. Although recent advances have been made in crucial aspects of the process, such composting microbiology, improvements are needed in process monitoring. Therefore, specific problems related to food waste composting, such as the presence of impurities, are thoroughly analysed in this study. In addition, environmental impacts related to food waste composting, such as emissions of greenhouse gases and odours, are discussed. Finally, the use of food waste compost in soil bioremediation is discussed in detail. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Hydrologic data for a subsurface waste-injection site at Mulberry, Florida; 1972-77

    USGS Publications Warehouse

    Wilson, William Edward; Parsons, David C.; Spechler, R.M.

    1979-01-01

    Since October 1972, industrial liquid waste has been injected into a brine aquifer of limestone and dolomite in Mulberry, FL., at a depth of more than 4,000 feet below land surface. During 1977, the injection rate was about 8.8 million gallons per month. To determine what effect the injected waste has on the ground-water body, water levels have been measured and water samples collected from two monitor wells that tap different permeable zones above the injection zone, and from a satellite monitor well that taps the injection zone. The monitor wells are in the annulus of the injection well, and the satellite monitor well is 2,291 feet from the injection well. This report updates previous data reports and includes all hydrologic data collected by the U.S. Geological Survey during 1972-77. Included is a table of well-construction data, a graph showing the volume of waste injected each month, and hydrographs of the annulus monitor wells and the satellite monitor well. (Woodard-USGS)

  3. Two High Throughput Screen Assays for Measurement of TNF-α in THP-1 Cells

    PubMed Central

    Leister, Kristin P; Huang, Ruili; Goodwin, Bonnie L; Chen, Andrew; Austin, Christopher P; Xia, Menghang

    2011-01-01

    Tumor Necrosis Factor-α (TNF-α), a secreted cytokine, plays an important role in inflammatory diseases and immune disorders, and is a potential target for drug development. The traditional assays for detecting TNF-α, enzyme linked immunosorbent assay (ELISA) and radioimmunoassay, are not suitable for the large size compound screens. Both assays suffer from a complicated protocol, multiple plate wash steps and/or excessive radioactive waste. A simple and quick measurement of TNF-α production in a cell based assay is needed for high throughput screening to identify the lead compounds from the compound library. We have developed and optimized two homogeneous TNF-α assays using the HTRF (homogeneous time resolved fluorescence) and AlphaLISA assay formats. We have validated the HTRF based TNF-α assay in a 1536-well plate format by screening a library of 1280 pharmacologically active compounds. The active compounds identified from the screen were confirmed in the AlphaLISA TNF-α assay using a bead-based technology. These compounds were also confirmed in a traditional ELISA assay. From this study, several beta adrenergic agonists have been identified as TNF-α inhibitors. We also identified several novel inhibitors of TNF-α, such as BTO-1, CCG-2046, ellipticine, and PD 169316. The results demonstrated that both homogeneous TNF-α assays are robust and suitable for high throughput screening. PMID:21643507

  4. Cardiac troponin T measured with a highly sensitive assay for diagnosis and monitoring of heart injury in chronic Chagas disease.

    PubMed

    Saravia, Silvia Gilka Muñoz; Haberland, Annekathrin; Bartel, Sabine; Araujo, Raul; Valda, Gregorio; Reynaga, Diana Duchen; Ramirez, Ivan Diaz; Borges, Adrian C; Wallukat, Gerd; Schimke, Ingolf

    2011-02-01

    Chronic Chagas disease (15 million patients; annual incidence, 40, 000 patients; annual mortality, 12 ,500 patients) is the most serious parasitic disease in Latin America. Between 10 and 30 years after infection, 30% of patients with Chagas disease develop heart injury, which is the main reason for its high mortality. Consequently, frequent cardiac diagnostics are required for patients with Chagas disease. To minimize time-intensive and cost-intensive diagnostics, such as electrocardiography, echocardiography, and radiologic imaging, we tested the effect of measuring serum cardiac troponin T (cTnT) with a highly sensitive assay. To indicate the pathophysiologic background for cTnT release in Chagas heart injury, inflammation markers, such as C-reactive protein and interleukin 6, were measured in parallel. Serum cTnT was measured in 26 healthy subjects and in 179 patients with chronic Chagas disease who were asymptomatic (indeterminate stage, n  =  86), who were suffering from cardiomyopathy with or without megacolon (n  =  71), or who were suffering from megacolon exclusively (n  =  22). Serum cTnT was significantly higher in patients with cardiomyopathy with or without megacolon than in healthy subjects, asymptomatic subjects, and patients with megacolon, and the cTnT value was correlated with the severity of the cardiomyopathy. The lower limit of detection for the highly sensitive assay (3 ng/L) was best at distinguishing patients with, and without, heart injury. C-reactive protein and interleukin 6 were found to parallel cTnT changes in both the different Chagas groups and the cardiomyopathy groups separated by disease severity. Highly sensitive cTnT measurement has the potential to contribute to diagnosis and monitoring of heart injury in patients with chronic Chagas disease. The highly sensitive assay of cTnT release seems to be related to Chagas heart disease-specific inflammation.

  5. Development of at-line assay to monitor charge variants of MAbs during production.

    PubMed

    St Amand, M M; Ogunnaike, B A; Robinson, A S

    2014-01-01

    One major challenge currently facing the biopharmaceutical industry is to understand how MAb microheterogeneity affects therapeutic efficacy, potency, immunogenicity, and clearance. MAb micro-heterogeneity can result from post-translational modifications such as sialylation, galactosylation, C-terminal lysine cleavage, glycine amidation, and tryptophan oxidation, each of which can generate MAb charge variants; such heterogeneity can affect pharmacokinetics (PK) considerably. Implementation of appropriate on-line quality control strategies may help to regulate bioprocesses, thus enabling more homogenous material with desired post-translational modifications and PK behavior. However, one major restriction to implementation of quality control strategies is the availability of techniques for obtaining on-line or at-line measurements of these attributes. In this work, we describe the development of an at-line assay to separate MAb charge variants in near real-time, which could ultimately be used to implement on-line quality control strategies for MAb production. The assay consists of a 2D-HPLC method with sequential in-line Protein A and WCX-10 HPLC column steps. To perform the 2D-HPLC assay at-line, the two columns steps were integrated into a single method using a novel system configuration that allowed parallel flow over column 1 or column 2 or sequential flow from column 1 to column 2. A bioreactor system was also developed such that media samples could be removed automatically from bioreactor vessels during production and delivered to the 2D-HPLC for analysis. With this at-line HPLC assay, we have demonstrated that MAb microheterogeneity occurs throughout the cell cycle whether the host cell line is grown under different or the same nominal culture conditions. © 2013 American Institute of Chemical Engineers.

  6. Plasmid transfection in bovine cells: Optimization using a realtime monitoring of green fluorescent protein and effect on gene reporter assay.

    PubMed

    Osorio, Johan S; Bionaz, Massimo

    2017-08-30

    Gene reporter technology (GRT) has opened several new avenues for monitoring biological events including the activation of transcription factors, which are central to the study of nutrigenomics. However, this technology relies heavily on the insertion of foreign plasmid DNA into the nuclei of cells (i.e., transfection), which can be very challenging and highly variable among cell types. The objective of this study was to investigate the optimal conditions to generate reliable GRT assay data on bovine immortalized cell lines, Madin Darby Bovine Kidney (MDBK) and bovine mammary epithelial alveolar (MACT) cells. Results are reported for two experiments. In Experiment 1, using 96 well-plate and a robotic inverted fluorescent microscope, we compared transfection efficiency among commercially available transfection reagents (TR) Lipofectamine® 3000 (Lipo3), Lipofectamine® LTX (LipoLTX), and TransIT-X2® (TransX2), three doses of TR (i.e., 0.15, 0.3, and 0.4μL/well), and three doses of Green Fluorescent Protein plasmid DNA (i.e., 10, 25, and 50ng/well). Transfection efficiency and mortality rate were analyzed using CellProfiler software. Transfection efficiency increased until the end of the experiment (20h post-transfection) at which point MACT had greater transfection than MDBK cells (16.3% vs. 2.2%). It is unclear the reason for the low transfection in MDBK cells. Maximal transfection efficiency was obtained with 0.3μL/well of LipoLTX plus 25ng/well of plasmid DNA (ca. 29.5±1.9%) and 0.15μL/well of LipoLTX plus 25ng/well of plasmid DNA (ca. 4.0±0.4%) for MACT and MDBK cells, respectively. The higher amount of TR and DNA was generally associated with higher cell mortality. Using high, medium, and low transfection efficiency conditions determined in Experiment 1, we performed a GRT assay for peroxisome proliferator-activated response element (PPRE) luciferase in MACT and MDBK cells treated with 10nM or 100nM of synthetic Peroxisome Proliferator-activated Receptor

  7. 2012 Annual Industrial Wastewater Reuse Report for the Idaho National Laboratory Site’s Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mike Lewis

    2013-02-01

    This report describes conditions, as required by the state of Idaho Industrial Wastewater Reuse Permit (WRU-I-0160-01, formerly LA 000160 01), for the wastewater reuse site at the Idaho National Laboratory Site’s Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond from November 1, 2011 through October 31, 2012. The report contains the following information: • Facility and system description • Permit required effluent monitoring data and loading rates • Groundwater monitoring data • Status of special compliance conditions • Discussion of the facility’s environmental impacts During the 2012 reporting year, an estimated 11.84 million gallons of wastewater weremore » discharged to the Industrial Waste Ditch and Pond which is well below the permit limit of 17 million gallons per year. The concentrations of all permit-required analytes in the samples from the down gradient monitoring wells were below the Ground Water Quality Rule Primary and Secondary Constituent Standards.« less

  8. 2013 Annual Industrial Wastewater Reuse Report for the Idaho National Laboratory Site’s Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mike Lewis

    This report describes conditions, as required by the state of Idaho Industrial Wastewater Reuse Permit (WRU-I-0160-01, formerly LA 000160 01), for the wastewater reuse site at the Idaho National Laboratory Site’s Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond from November 1, 2012 through October 31, 2013. The report contains the following information: • Facility and system description • Permit required effluent monitoring data and loading rates • Groundwater monitoring data • Status of special compliance conditions • Discussion of the facility’s environmental impacts During the 2013 reporting year, an estimated 9.64 million gallons of wastewater weremore » discharged to the Industrial Waste Ditch and Pond which is well below the permit limit of 17 million gallons per year. The concentrations of all permit-required analytes in the samples from the down gradient monitoring wells were below the applicable Idaho Department of Environmental Quality’s groundwater quality standard levels.« less

  9. 2014 Annual Industrial Wastewater Reuse Report for the Idaho National Laboratory Site’s Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lewis, Mike

    This report describes conditions, as required by the state of Idaho Industrial Wastewater Reuse Permit (WRU-I-0160-01, formerly LA 000160 01), for the wastewater reuse site at the Idaho National Laboratory Site’s Materials and Fuels Complex Industrial Waste Ditch and Industrial Waste Pond from November 1, 2013 through October 31, 2014. The report contains the following information; Facility and system description; Permit required effluent monitoring data and loading rates; Groundwater monitoring data; Status of special compliance conditions; Noncompliance issues; and Discussion of the facility’s environmental impacts During the 2014 reporting year, an estimated 10.11 million gallons of wastewater were discharged tomore » the Industrial Waste Ditch and Pond which is well below the permit limit of 17 million gallons per year. The concentrations of all permit-required analytes in the samples from the down gradient monitoring wells were below the applicable Idaho Department of Environmental Quality’s groundwater quality standard levels.« less

  10. Waste inspection tomography (WIT)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bernardi, R.T.

    1996-12-31

    WIT is a self-sufficient mobile semitrailer for nondestructive evaluation and nondestructive assay of nuclear waste drums using x-ray and gamma-ray tomography. The recently completed Phase I included the design, fabrication, and initial testing of all WIT subsystems installed on-board the trailer. Initial test results include 2 MeV digital radiography, computed tomography, Anger camera imaging, single photon emission computed tomography, gamma-ray spectroscopy, collimated gamma scanning, and active and passive computed tomography using a 1.4 mCi source of {sup 166}Ho. These techniques were initially demonstrated on a 55-gallon phantom drum with 3 simulated waste matrices of combustibles, heterogeneous metals, and cement usingmore » check sources of gamma active isotopes such as {sup 137}Cs and {sup 133}Ba with 9-250 {mu}Ci activities. Waste matrix identification, isotopic identification, and attenuation-corrected gamma activity determination were demonstrated nondestructively and noninvasively in Phase I. Currently ongoing Phase II involves DOE site field test demonstrations at LLNL, RFETS, and INEL with real nuclear waste drums. Current WIT experience includes 55 gallon drums of cement, graphite, sludge, glass, metals, and combustibles. Thus far WIT has inspected drums with 0-20 gms of {sup 239}Pu.« less

  11. 2002 Hyperspectral Analysis of Hazardous Waste Sites on the Savannah River Site

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gladden, J.B.

    2003-08-28

    Hazardous waste site inspection is a labor intensive, time consuming job, performed primarily on the ground using visual inspection and instrumentation. It is an expensive process to continually monitor hazardous waste and/or landfill sites to determine if they are maintaining their integrity. In certain instances, it may be possible to monitor aspects of the hazardous waste sites and landfills remotely. The utilization of multispectral data was suggested for the mapping of clays and iron oxides associated with contaminated groundwater, vegetation stress, and methane gas emissions (which require longer wavelength detectors). The Savannah River Site (SRS) near Aiken, S.C. is amore » United States Department of Energy facility operated by the Westinghouse Savannah River Company. For decades the SRS was responsible for developing weapons grade plutonium and other materials for the nation's nuclear defense. Hazardous waste was generated during this process. Waste storage site inspection is a particularly important issue at the SRS because there are over 100 hazardous waste sites scattered throughout the 300 mile complex making it difficult to continually monitor all of the facilities. The goal is to use remote sensing technology to identify surface anomalies on the hazardous waste sites as early as possible so that remedial work can take place rapidly to maintain the integrity of the storage sites. The anomalous areas are then targeted for intensive in situ human examination and measurement. During the 1990s, many of the hazardous waste sites were capped with protective layers of polyethelene sheeting and soil, and planted with bahia grass and/or centipede grass. This research investigated hyperspectral remote sensing technology to determine if it can be used to measure accurately and monitor possible indicators of change on vegetated hazardous waste sites. Specifically, it evaluated the usefulness of hyperspectral remote sensing to assess the condition of vegetation

  12. The comparison of fossil carbon fraction and greenhouse gas emissions through an analysis of exhaust gases from urban solid waste incineration facilities.

    PubMed

    Kim, Seungjin; Kang, Seongmin; Lee, Jeongwoo; Lee, Seehyung; Kim, Ki-Hyun; Jeon, Eui-Chan

    2016-10-01

    In this study, in order to understand accurate calculation of greenhouse gas emissions of urban solid waste incineration facilities, which are major waste incineration facilities, and problems likely to occur at this time, emissions were calculated by classifying calculation methods into 3 types. For the comparison of calculation methods, the waste characteristics ratio, dry substance content by waste characteristics, carbon content in dry substance, and (12)C content were analyzed; and in particular, CO2 concentration in incineration gases and (12)C content were analyzed together. In this study, 3 types of calculation methods were made through the assay value, and by using each calculation method, emissions of urban solid waste incineration facilities were calculated then compared. As a result of comparison, with Calculation Method A, which used the default value as presented in the IPCC guidelines, greenhouse gas emissions were calculated for the urban solid waste incineration facilities A and B at 244.43 ton CO2/day and 322.09 ton CO2/day, respectively. Hence, it showed a lot of difference from Calculation Methods B and C, which used the assay value of this study. It is determined that this was because the default value as presented in IPCC, as the world average value, could not reflect the characteristics of urban solid waste incineration facilities. Calculation Method B indicated 163.31 ton CO2/day and 230.34 ton CO2/day respectively for the urban solid waste incineration facilities A and B; also, Calculation Method C indicated 151.79 ton CO2/day and 218.99 ton CO2/day, respectively. This study intends to compare greenhouse gas emissions calculated using (12)C content default value provided by the IPCC (Intergovernmental Panel on Climate Change) with greenhouse gas emissions calculated using (12)C content and waste assay value that can reflect the characteristics of the target urban solid waste incineration facilities. Also, the concentration and (12)C content

  13. CPTAC Assay Portal: a repository of targeted proteomic assays

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Whiteaker, Jeffrey R.; Halusa, Goran; Hoofnagle, Andrew N.

    2014-06-27

    To address these issues, the Clinical Proteomic Tumor Analysis Consortium (CPTAC) of the National Cancer Institute (NCI) has launched an Assay Portal (http://assays.cancer.gov) to serve as a public repository of well-characterized quantitative, MS-based, targeted proteomic assays. The purpose of the CPTAC Assay Portal is to facilitate widespread adoption of targeted MS assays by disseminating SOPs, reagents, and assay characterization data for highly characterized assays. A primary aim of the NCI-supported portal is to bring together clinicians or biologists and analytical chemists to answer hypothesis-driven questions using targeted, MS-based assays. Assay content is easily accessed through queries and filters, enabling investigatorsmore » to find assays to proteins relevant to their areas of interest. Detailed characterization data are available for each assay, enabling researchers to evaluate assay performance prior to launching the assay in their own laboratory.« less

  14. Subseabed storage of radioactive waste

    NASA Astrophysics Data System (ADS)

    Bell, Peter M.

    The subject of the storage of nuclear wastes products incites emotional responses from the public, and thus the U.S. Subseabed Disposal Program will have to make a good case for waste storage beneath the ocean floor. The facts attendant, however, describe circumstances necessitating cool-headed analysis to achieve a solution to the growing nuclear waste problem. Emotion aside, a good case indeed is being made for safe disposal beneath the ocean floor.The problems of nuclear waste storage are acute. A year ago, U.S. military weapons production had accumulated over seventy-five million gallons of high-level radioactive liquid waste; solid wastes, such as spent nuclear fuel rods from reactors, amounted to more than 12,000 tons. These wastes are corrosive and will release heat for 1000 years or more. The wastes will remain dangerously radioactive for a period of 10,000 years. There are advantages in storing the wastes on land, in special underground repositories, or on the surface. These include the accessibility to monitor the waste and the possibility of taking action should a container rupture occur, and thus the major efforts to determine suitable disposal at this time are focused on land-based storage. New efforts, not to be confused with ocean dumping practices of the past, are demonstrating that waste containers isolated in the clays and sediments of the ocean floor may be superior (Environ. Sci. Tech., 16, 28A-37A 1982).

  15. Municipal solid-waste disposal and ground-water quality in a coastal environment, west-central Florida

    USGS Publications Warehouse

    Fernandez, Mario

    1983-01-01

    Solid waste is defined along with various methods of disposal and the hydrogeologic factors to be considered when locating land-fills is presented. Types of solid waste, composition, and sources are identified. Generation of municipal solid waste in Florida has been estimated at 4.5 pounds per day per person or about 7.8 million tons per year. Leachate is generated when precipitation and ground water percolate through the waste. Gases, mainly carbon dioxide and methane, are also produced. Leachate generally contains high concentrations of dissolved organic and inorganic matter. The two typical hydrogeologic conditions in west-central Florida are (1) permeable sand overlying clay and limestone and (2) permeable sand overlying limestone. These conditions are discussed in relation to leachate migration. Factors in landfill site selection are presented and discussed, followed by a discussion on monitoring landfills. Monitoring of landfills includes the drilling of test holes, measuring physical properties of the corings, installation of monitoring wells, and water-quality monitoring. (USGS)

  16. Facilitating the improved management of waste in South Africa through a national waste information system

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Godfrey, Linda

    2008-07-01

    Developing a waste information system (WIS) for a country is more than just about collecting routine data on waste; it is about facilitating the improved management of waste by providing timely, reliable information to the relevant role-players. It is a means of supporting the waste governance challenges facing South Africa - challenges ranging from strategic waste management issues at national government to basic operational challenges at local government. The paper addresses two hypotheses. The first is that the identified needs of government can provide a platform from which to design a national WIS framework for a developing country such asmore » South Africa, and the second is that the needs for waste information reflect greater, currently unfulfilled challenges in the sustainable management of waste. Through a participatory needs analysis process, it is shown that waste information is needed by the three spheres of government, to support amongst others, informed planning and decision-making, compliance monitoring and enforcement, community participation through public access to information, human, infrastructure and financial resource management and policy development. These needs for waste information correspond closely with key waste management challenges currently facing the country. A shift in governments approach to waste, in line with national and international policy, is evident from identified current and future waste information needs. However, the need for information on landfilling remains entrenched within government, possibly due to the poor compliance of landfill sites in South Africa and the problems around the illegal disposal of both general and hazardous waste.« less

  17. Radioactive Waste Management in A Hospital

    PubMed Central

    Khan, Shoukat; Syed, AT; Ahmad, Reyaz; Rather, Tanveer A.; Ajaz, M; Jan, FA

    2010-01-01

    Most of the tertiary care hospitals use radioisotopes for diagnostic and therapeutic applications. Safe disposal of the radioactive waste is a vital component of the overall management of the hospital waste. An important objective in radioactive waste management is to ensure that the radiation exposure to an individual (Public, Radiation worker, Patient) and the environment does not exceed the prescribed safe limits. Disposal of Radioactive waste in public domain is undertaken in accordance with the Atomic Energy (Safe disposal of radioactive waste) rules of 1987 promulgated by the Indian Central Government Atomic Energy Act 1962. Any prospective plan of a hospital that intends using radioisotopes for diagnostic and therapeutic procedures needs to have sufficient infrastructural and manpower resources to keep its ambient radiation levels within specified safe limits. Regular monitoring of hospital area and radiation workers is mandatory to assess the quality of radiation safety. Records should be maintained to identify the quality and quantity of radioactive waste generated and the mode of its disposal. Radiation Safety officer plays a key role in the waste disposal operations. PMID:21475524

  18. Radioactive waste management in a hospital.

    PubMed

    Khan, Shoukat; Syed, At; Ahmad, Reyaz; Rather, Tanveer A; Ajaz, M; Jan, Fa

    2010-01-01

    Most of the tertiary care hospitals use radioisotopes for diagnostic and therapeutic applications. Safe disposal of the radioactive waste is a vital component of the overall management of the hospital waste. An important objective in radioactive waste management is to ensure that the radiation exposure to an individual (Public, Radiation worker, Patient) and the environment does not exceed the prescribed safe limits. Disposal of Radioactive waste in public domain is undertaken in accordance with the Atomic Energy (Safe disposal of radioactive waste) rules of 1987 promulgated by the Indian Central Government Atomic Energy Act 1962. Any prospective plan of a hospital that intends using radioisotopes for diagnostic and therapeutic procedures needs to have sufficient infrastructural and manpower resources to keep its ambient radiation levels within specified safe limits. Regular monitoring of hospital area and radiation workers is mandatory to assess the quality of radiation safety. Records should be maintained to identify the quality and quantity of radioactive waste generated and the mode of its disposal. Radiation Safety officer plays a key role in the waste disposal operations.

  19. Rapid assessment of antibody-induced ricin neutralization by employing a novel functional cell-based assay.

    PubMed

    Gal, Yoav; Alcalay, Ron; Sabo, Tamar; Noy-Porat, Tal; Epstein, Eyal; Kronman, Chanoch; Mazor, Ohad

    2015-09-01

    Ricin is one of the most potent and lethal toxins known against which there is no available antidote. Currently, the most promising countermeasures against the toxin are based on neutralizing antibodies elicited by active vaccination or administered passively. A cell-based assay is widely applied for the primary screening and evaluation of anti-ricin antibodies, yet such assays are usually time-consuming (18-72 h). Here, we report of a novel assay to monitor ricin activity, based on HeLa cells that stably express the rapidly-degraded ubiquitin-luciferase (Ub-FL, half-life of 2 min). Ricin-induced arrest of protein synthesis could be quantified within 3 to 6h post intoxication (IC90 of 300 and 100 ng/ml, respectively). Furthermore, by stabilizing the intracellular levels of Ub-FL in the last hour of the assay, a 3-fold increase in the assay sensitivity was attained. We applied this assay to monitor the efficacy of a ricin holotoxin-based vaccine by measuring the formation of neutralizing antibodies throughout the immunization course. The potency of anti-ricin monoclonal antibodies (directed to either subunit of the toxin) could also be easily and accurately measured in this assay format. Owing to its simplicity, this assay may be implemented for high-throughput screening of ricin-neutralizing antibodies and for identification of small-molecule inhibitors of the toxin, as well as other ribosome-inactivating toxins. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. PERFORMANCE MONITORING FOR NATURAL ATTENUATION OF CONTAMINANTS IN AQUIFERS

    EPA Science Inventory

    Monitoring of hazardous waste sites serves a variety of very distinct purposes. The initial phase of monitoring is designed to determine whether there is a problem at a site (remedial investigation). If a problem exists, the next phase of monitoring is designed to select a reme...

  1. Evaluation of performance across the dynamic range of the Abbott RealTime HIV-1 assay as compared to VERSANT HIV-1 RNA 3.0 and AMPLICOR HIV-1 MONITOR v1.5 using serial dilutions of 39 group M and O viruses.

    PubMed

    Swanson, Priscilla; Huang, Shihai; Abravaya, Klara; de Mendoza, Carmen; Soriano, Vincent; Devare, Sushil G; Hackett, John

    2007-04-01

    Performance of the Abbott m2000 instrument system and the Abbott RealTime HIV-1 assay was evaluated using a panel of 37 group M (subtypes A-D, F, G, CRF01_AE, CRF02_AG and unique recombinant forms) and 2 group O virus isolates. Testing was performed on 273 sample dilutions and compared to VERSANT HIV-1 RNA 3.0 (bDNA) and AMPLICOR HIV-1 MONITOR v1.5 (Monitor v1.5) test results. RealTime HIV-1, bDNA, and Monitor v1.5 tests quantified 87%, 78%, and 81% of samples, respectively. RealTime HIV-1 detected an additional 31 samples at < 40 copies/mL. For group M, RealTime HIV-1 dilution profiles and viral loads were highly correlated with bDNA and Monitor v1.5 values; 87% and 89% of values were within 0.5 log(10) copies/mL. In contrast, the group O viruses were not detected by Monitor v1.5 and were substantially underquantified by approximately 2 log(10) copies/mL in bDNA relative to the RealTime HIV-1 assay. Sequence analysis revealed that RealTime HIV-1 primer/probe binding sites are highly conserved and exhibit fewer nucleotide mismatches relative to Monitor v1.5. The automated m2000 system and RealTime HIV-1 assay offer the advantages of efficient sample processing and throughput with reduced "hands-on" time while providing improved sensitivity, expanded dynamic range and reliable quantification of genetically diverse HIV-1 strains.

  2. Implementation of anion-receptor macrocycles in supramolecular tandem assays for enzymes involving nucleotides as substrates, products, and cofactors.

    PubMed

    Florea, Mara; Nau, Werner M

    2010-03-07

    A supramolecular tandem assay for direct continuous monitoring of nucleotide triphosphate-dependent enzymes such as potato apyrase is described. The underlying principle of the assay relies on the use of anion-receptor macrocycles in combination with fluorescent dyes as reporter pairs. A combinatorial approach was used to identify two complementary reporter pairs, i.e. an amino-gamma-cyclodextrin with 2-anilinonaphtalene-6-sulfonate (ANS) as dye (fluorescence enhancement factor of 17 upon complexation) and a polycationic cyclophane with 8-hydroxy-1,3,6-pyrene trisulfonate (HPTS) as dye (fluorescence decrease by a factor of more than 2000), which allow the kinetic monitoring of potato apyrase activity at different ATP concentration ranges (microM and mM) with different types of photophysical responses (switch-ON and switch-OFF). Competitive fluorescence titrations revealed a differential binding of ATP (strongest competitor) versus ADP and AMP, which constitutes the prerequisite for monitoring enzymatic conversions (dephosphorylation or phosphorylation) involving nucleotides. The assay was tested for different enzyme and substrate concentrations and exploited for the screening of activating additives, namely divalent transition metal ions (Ni(2+), Mg(2+), Mn(2+), and Ca(2+)). The transferability of the assay could be demonstrated by monitoring the dephosphorylation of other nucleotide triphosphates (GTP, TTP, and CTP).

  3. RFID technology for hazardous waste management and tracking.

    PubMed

    Namen, Anderson Amendoeira; Brasil, Felipe da Costa; Abrunhosa, Jorge José Gouveia; Abrunhosa, Glaucia Gomes Silva; Tarré, Ricardo Martinez; Marques, Flávio José Garcia

    2014-09-01

    The illegal dumping of hazardous waste is one of the most concerning occurrences related to illegal waste activities. The waste management process is quite vulnerable, especially when it comes to assuring the right destination for the delivery of the hazardous waste. The purpose of this paper is to present a new system design and prototype for applying the RFID technology so as to guarantee the correct destination for the hazardous waste delivery. The aim of this innovative approach, compared with other studies that employ the same technology to the waste disposal process, is to focus on the certification that the hazardous waste will be delivered to the right destination site and that no inappropriate disposal will occur in the transportation stage. These studies were carried out based on data collected during visits to two hazardous waste producer companies in Brazil, where the material transportation and delivery to a company in charge of the waste disposal were closely monitored. © The Author(s) 2014.

  4. 40 CFR 257.24 - Detection monitoring program.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Disposal Units Ground-Water Monitoring and Corrective Action § 257.24 Detection monitoring program. (a... unit; (ii) The mobility, stability, and persistence of waste constituents or their reaction products in... constituents, and reaction products in the ground water; and (iv) The concentration or values and coefficients...

  5. Waste stream recycling: Its effect on water quality

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cornwell, D.A.; Lee, R.G.

    1994-11-01

    Waste streams recycled to the influent of a water treatment plant typically contain contaminants at concentrations that are of concern. These contaminants may include giardia and Cryptosporidium, trihalomethanes, manganese, and assimilable organic carbon. This research shows that proper management--treatment, equalization, and monitoring--of the waste streams can render them suitable for recycling in many situations.

  6. In vitro immunotherapy potency assays using real-time cell analysis

    PubMed Central

    Cerignoli, Fabio; Abassi, Yama A.; Lamarche, Brandon J.; Guenther, Garret; Santa Ana, David; Guimet, Diana; Zhang, Wen; Zhang, Jing

    2018-01-01

    A growing understanding of the molecular interactions between immune effector cells and target tumor cells, coupled with refined gene therapy approaches, are giving rise to novel cancer immunotherapeutics with remarkable efficacy in the clinic against both solid and liquid tumors. While immunotherapy holds tremendous promise for treatment of certain cancers, significant challenges remain in the clinical translation to many other types of cancers and also in minimizing adverse effects. Therefore, there is an urgent need for functional potency assays, in vitro and in vivo, that could model the complex interaction of immune cells with tumor cells and can be used to rapidly test the efficacy of different immunotherapy approaches, whether it is small molecule, biologics, cell therapies or combinations thereof. Herein we report the development of an xCELLigence real-time cytolytic in vitro potency assay that uses cellular impedance to continuously monitor the viability of target tumor cells while they are being subjected to different types of treatments. Specialized microtiter plates containing integrated gold microelectrodes enable the number, size, and surface attachment strength of adherent target tumor cells to be selectively monitored within a heterogeneous mixture that includes effector cells, antibodies, small molecules, etc. Through surface-tethering approach, the killing of liquid cancers can also be monitored. Using NK92 effector cells as example, results from RTCA potency assay are very well correlated with end point data from image-based assays as well as flow cytometry. Several effector cells, i.e., PBMC, NK, CAR-T were tested and validated as well as biological molecules such as Bi-specific T cell Engagers (BiTEs) targeting the EpCAM protein expressed on tumor cells and blocking antibodies against the immune checkpoint inhibitor PD-1. Using the specifically designed xCELLigence immunotherapy software, quantitative parameters such as KT50 (the amount of time

  7. In Vitro Assays for Assessment of Androgenic and Estrogenic Activity of Defined Mixtures and Complex Environmental Samples

    EPA Science Inventory

    Point sources of endocrine active compounds to aquatic environments such as waste water treatment plants, pulp and paper mills, and animal feeding operations invariably contain complex mixtures of chemicals. The current study investigates the use of targeted in vitro assays des...

  8. Development of Isothermal Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Circovirus Type 2.

    PubMed

    Yang, Yang; Qin, Xiaodong; Sun, Yingjun; Cong, Guozheng; Li, Yanmin; Zhang, Zhidong

    2017-01-01

    Porcine circovirus virus type II (PCV2) is the etiology of postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis, nephropathy syndrome (PDNS), and necrotizing pneumonia. Rapid diagnosis tool for detection of PCV2 plays an important role in the disease control and eradication program. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PCV2 real-time RPA assay) and RPA combined with lateral flow dipstick (PCV2 RPA LFD assay) were developed targeting the PCV2 ORF2 gene. The results showed that the sensitivity of the PCV2 real-time RPA assay was 10 2 copies per reaction within 20 min at 37°C and the PCV2 RPA LFD assay had a detection limit of 10 2 copies per reaction in less than 20 min at 37°C. Both assays were highly specific for PCV2, with no cross-reactions with porcine circovirus virus type 1, foot-and-mouth disease virus, pseudorabies virus, porcine parvovirus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus. Therefore, the RPA assays provide a novel alternative for simple, sensitive, and specific identification of PCV2.

  9. Development of Isothermal Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Circovirus Type 2

    PubMed Central

    Yang, Yang; Qin, Xiaodong; Sun, Yingjun; Cong, Guozheng; Li, Yanmin

    2017-01-01

    Porcine circovirus virus type II (PCV2) is the etiology of postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis, nephropathy syndrome (PDNS), and necrotizing pneumonia. Rapid diagnosis tool for detection of PCV2 plays an important role in the disease control and eradication program. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PCV2 real-time RPA assay) and RPA combined with lateral flow dipstick (PCV2 RPA LFD assay) were developed targeting the PCV2 ORF2 gene. The results showed that the sensitivity of the PCV2 real-time RPA assay was 102 copies per reaction within 20 min at 37°C and the PCV2 RPA LFD assay had a detection limit of 102 copies per reaction in less than 20 min at 37°C. Both assays were highly specific for PCV2, with no cross-reactions with porcine circovirus virus type 1, foot-and-mouth disease virus, pseudorabies virus, porcine parvovirus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus. Therefore, the RPA assays provide a novel alternative for simple, sensitive, and specific identification of PCV2. PMID:28424790

  10. Novel heparan sulfate assay by using automated high-throughput mass spectrometry: application to monitoring and screening for mucopolysaccharidoses

    PubMed Central

    Shimada, Tsutomu; Kelly, Joan; LaMarr, William A; van Vlies, Naomi; Yasuda, Eriko; Mason, Robert W.; Mackenzie, William; Kubaski, Francyne; Giugliani, Roberto; Chinen, Yasutsugu; Yamaguchi, Seiji; Suzuki, Yasuyuki; Orii, Kenji E.; Fukao, Toshiyuki; Orii, Tadao; Tomatsu, Shunji

    2014-01-01

    assay, indicating that HT-MS/MS may be feasible for diagnosis, monitoring, and newborn screening of MPS. PMID:25092413

  11. Measuring oxidative damage to DNA and its repair with the comet assay.

    PubMed

    Collins, Andrew R

    2014-02-01

    Single cell gel electrophoresis, or the comet assay, was devised as a sensitive method for detecting DNA strand breaks, at the level of individual cells. A simple modification, incorporating a digestion of DNA with a lesion-specific endonuclease, makes it possible to measure oxidised bases. With the inclusion of formamidopyrimidine DNA glycosylase to recognise oxidised purines, or Nth (endonuclease III) to detect oxidised pyrimidines, the comet assay has been used extensively in human biomonitoring to monitor oxidative stress, usually in peripheral blood mononuclear cells. There is evidence to suggest that the enzymic approach is more accurate than chromatographic methods, when applied to low background levels of base oxidation. However, there are potential problems of over-estimation (because the enzymes are not completely specific) or under-estimation (failure to detect lesions that are close together). Attempts have been made to improve the inter-laboratory reproducibility of the comet assay. In addition to measuring DNA damage, the assay can be used to monitor the cellular or in vitro repair of strand breaks or oxidised bases. It also has applications in assessing the antioxidant status of cells. In its various forms, the comet assay is now an invaluable tool in human biomonitoring and genotoxicity testing. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Polymeric assay film for direct colorimetric detection

    DOEpatents

    Charych, Deborah; Nagy, Jon; Spevak, Wayne

    2002-01-01

    A lipid bilayer with affinity to an analyte, which directly signals binding by a changes in the light absorption spectra. This novel assay means and method has special applications in the drug development and medical testing fields. Using a spectrometer, the system is easily automated, and a multiple well embodiment allows inexpensive screening and sequential testing. This invention also has applications in industry for feedstock and effluent monitoring.

  13. Polymeric assay film for direct colorimetric detection

    DOEpatents

    Charych, Deborah; Nagy, Jon; Spevak, Wayne

    1999-01-01

    A lipid bilayer with affinity to an analyte, which directly signals binding by a changes in the light absorption spectra. This novel assay means and method has special applications in the drug development and medical testing fields. Using a spectrometer, the system is easily automated, and a multiple well embodiment allows inexpensive screening and sequential testing. This invention also has applications in industry for feedstock and effluent monitoring.

  14. Aptima HIV-1 Quant Dx--A fully automated assay for both diagnosis and quantification of HIV-1.

    PubMed

    Nair, Sangeetha Vijaysri; Kim, Hee Cheol; Fortunko, Jacqueline; Foote, Tracy; Peling, Tashi; Tran, Cuong; Nugent, Charles Thomas; Joo, Sunghae; Kang, Youna; Wilkins, Bana; Lednovich, Kristen; Worlock, Andrew

    2016-04-01

    Separate assays are available for diagnosis and viral load (VL) monitoring of HIV-1. Studies have shown that using a single test for both confirmatory diagnosis and VL increases linkage to care. To validate a single assay for both diagnosis and VL monitoring of HIV-1 on the fully automated Panther platform. Validate the assay by assessing specificity, sensitivity, subtype detection, seroconversion, reproducibility and linearity. Also assess diagnostic agreement with the Procleix(®) Ultrio Elite™ discriminatory assay (Procleix), and agreement of VL results (method comparison) with Ampliprep/COBAS TaqMan HIV-1 version 2.0 (CAP/CTM), using clinical samples. The assay was specific (100%) and sensitive with a 95% limit of detection of 12 copies/mL with the 3rd WHO standards. Aptima detected HIV in seroconversion panels 6 and 11 days before p24 antigen and antibody tests, respectively. Diagnostic agreement with Procleix, was 100%. Regression analysis showed good agreement of VL results between Aptima and CAP/CTM with a slope of 1.02, intercept of 0.07, and correlation coefficient (R(2)) of 0.97. Aptima was more sensitive than CAP/CTM. Equivalent quantification was seen on testing clinical samples and isolates belonging to HIV group M, N, O and P and commercially available subtype panels. Assay results were linear (R(2) 0.9994) with standard deviation of <0.17 log copies across assay range. The good specificity, sensitivity, precision, subtype performance and clinical agreement with other assays demonstrated by Aptima combined with the complete automation provided by the Panther platform makes Aptima a good candidate for both VL monitoring and diagnosis of HIV-1. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  15. Radiological Characterization Methodology of INEEL Stored RH-TRU Waste from ANL-E

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rajiv N. Bhatt

    2003-02-01

    An Acceptable Knowledge (AK)-based radiological characterization methodology is being developed for RH TRU waste generated from ANL-E hot cell operations performed on fuel elements irradiated in the EBR-II reactor. The methodology relies on AK for composition of the fresh fuel elements, their irradiation history, and the waste generation and collection processes. Radiological characterization of the waste involves the estimates of the quantities of significant fission products and transuranic isotopes in the waste. Methods based on reactor and physics principles are used to achieve these estimates. Because of the availability of AK and the robustness of the calculation methods, the AK-basedmore » characterization methodology offers a superior alternative to traditional waste assay techniques. Using this methodology, it is shown that the radiological parameters of a test batch of ANL-E waste is well within the proposed WIPP Waste Acceptance Criteria limits.« less

  16. Application of the Attagene FACTORIAL™ assay to ...

    EPA Pesticide Factsheets

    Bioassays can be used to evaluate the integrated effects of complex mixtures from both known and unidentified contaminants present in environmental samples. However, such bio-monitoring approaches have typically focused only on one or a few pathways (e.g. estrogen receptor, androgen receptor) despite the fact that the chemicals in a mixture may exhibit a range of biological activities. High-throughput screening approaches that can rapidly assess samples for a broad diversity of biological activities offer a means to provide a more comprehensive characterization of complex mixtures. The Attagene FactorialTM platform is a high-throughput, cell based assay utilized by US EPA’s ToxCast Program, which provides high-content assessment of over 90 different gene regulatory pathways and all 48 human nuclear receptors (NRs). This assay has previously been used in a preliminary screening of surface water extracts from sites across the Great Lakes. In the current study, surface waters samples from 38 sites were collected, extracted, and screened through the Factorial assay as part of a USGS nationwide stream assessment. All samples were evaluated in a six point, 3-fold dilution series and analyzed using the ToxCast Data Pipeline (TCPL) to generate dose-response curves and corresponding half-maximal activity concentration (AC50) estimates. A total of 27 assay endpoints responded to extracts from one or more sites, with up to 14 assays active for a single extract. The four

  17. Comparison between three quantitative assays in patients with chronic hepatitis C and their relevance in the prediction of response to therapy.

    PubMed

    Pradat, P; Chossegros, P; Bailly, F; Pontisso, P; Saracco, G; Sauleda, S; Thursz, M; Tillmann, H; Vlassopoulou, H; Alberti, A; Braconier, J H; Esteban, J I; Hadziyannis, S; Manns, M; Rizzetto, M; Thomas, H C; Trépo, C

    2000-05-01

    To compare three quantitative assays measuring viral load in patients with chronic hepatitis C and to determine their value in predicting response to interferon (IFN) therapy, we analysed serum from 896 patients from eight European Centres using QUANTIPLEXtrade mark bDNA, MONITOR AMPLICORtrade mark and SUPERQUANTtrade mark assays. Analyses were performed on the same sample. Viral genotype was assessed using INNO-LiPA HCV II kits. Intercentre variations were observed that were related to the handling of specimens not processed and stored within 6 h of blood sampling. Among sera with optimal handling, a stronger correlation was observed between bDNA and SUPERQUANT (0.806) than between bDNA and MONITOR (0.677) and between MONITOR and SUPERQUANT (0.632). These discrepancies were greatest with genotype 2 (bDNA/SUPERQUANT= 0.772; bDNA/MONITOR=0. 456; SUPERQUANT/MONITOR= 0.299). This correlation was influenced by viraemia level and was better at lower viral loads. The proportion of sera with undetectable viral load was 15% with bDNA, 9.7% with MONITOR and 7.7% with SUPERQUANT. For the three measurements, the best cut-offs of sustained response to IFN treatment were located at their detection threshold. Among patients with viral load below the detection level, a sustained response was observed in 35% tested with bDNA, 38% with MONITOR and 80% with SUPERQUANT. Hence a stronger correlation was observed between bDNA and SUPERQUANT than between either of these assays and MONITOR. SUPERQUANT was the most sensitive assay and this greater sensitivity was associated with a better predictive value of treatment response.

  18. Semi-Tomographic Gamma Scanning Technique for Non-Destructive Assay of Radioactive Waste Drums

    NASA Astrophysics Data System (ADS)

    Gu, Weiguo; Rao, Kaiyuan; Wang, Dezhong; Xiong, Jiemei

    2016-12-01

    Segmented gamma scanning (SGS) and tomographic gamma scanning (TGS) are two traditional detection techniques for low and intermediate level radioactive waste drum. This paper proposes one detection method named semi-tomographic gamma scanning (STGS) to avoid the poor detection accuracy of SGS and shorten detection time of TGS. This method and its algorithm synthesize the principles of SGS and TGS. In this method, each segment is divided into annual voxels and tomography is used in the radiation reconstruction. The accuracy of STGS is verified by experiments and simulations simultaneously for the 208 liter standard waste drums which contains three types of nuclides. The cases of point source or multi-point sources, uniform or nonuniform materials are employed for comparison. The results show that STGS exhibits a large improvement in the detection performance, and the reconstruction error and statistical bias are reduced by one quarter to one third or less for most cases if compared with SGS.

  19. Real-time monitoring and control of the plasma hearth process

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Power, M.A.; Carney, K.P.; Peters, G.G.

    1996-05-01

    A distributed monitoring and control system is proposed for a plasma hearth, which will be used to decompose hazardous organic materials, encapsulate actinide waste in an obsidian-like slag, and reduce storage volume of actinide waste. The plasma hearth will be installed at ANL-West with the assistance of SAIC. Real-time monitoring of the off-gas system is accomplished using a Sun Workstation and embedded PCs. LabWindows/CVI software serves as the graphical user interface.

  20. A Proposal for Geologic Radioactive Waste Disposal Environmental Zero-State and Subsequent Monitoring Definition - First Lessons Learned from the French Environment Observatory - 13188

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Landais, Patrick; Leclerc, Elisabeth; Mariotti, Andre

    Obtaining a reference state of the environment before the beginning of construction work for a geological repository is essential as it will be useful for further monitoring during operations and beyond, thus keeping a memory of the original environmental state. The area and the compartments of the biosphere to be observed and monitored as well as the choice of the markers (e.g. bio-markers, biodiversity, quality of the environment, etc.) to be followed must be carefully selected. In parallel, the choice and selection of the environmental monitoring systems (i.e. scientific and technical criteria, social requirements) will be of paramount importance formore » the evaluation of the perturbations that could be induced during the operational phase of the repository exploitation. This paper presents learning points of the French environment observatory located in the Meuse/Haute-Marne that has been selected for studying the feasibility of the underground disposal of high level wastes in France. (authors)« less

  1. Vermicomposting of a lignocellulosic waste from olive oil industry: a pilot scale study.

    PubMed

    Benítez, E; Sainz, H; Melgar, R; Nogales, R

    2002-04-01

    The vermicomposting with Eisenia andrei of dry olive cake, a lignocellulosic waste produced during the extraction of olive oil, either alone or mixed with municipal biosolids, was studied in a nine-month pilot scale experiment. Number and biomass of earthworms and enzyme activities were periodically monitored and relevant properties of the final products were determined. In the assayed substrates, the total biomass of earthworms increased at the end of the experimental period between 9 and 12-fold respectively in comparison with the earthworm biomass initially inoculated. The increase in hydrolytic enzymes and overall microbial activity during the vermicomposting process indicated the biodegradation of the olive cake and resulted in the disappearance of the initial phytotoxicity of the substrate. However, the recalcitrant lignocellulosic nature of the dry olive cake prevented suitable humification during the vermicomposting process. For this reason, in addition to organic amendments, other management procedures should be considered.

  2. Monitoring Metal Pollution Levels in Mine Wastes around a Coal Mine Site Using GIS

    NASA Astrophysics Data System (ADS)

    Sanliyuksel Yucel, D.; Yucel, M. A.; Ileri, B.

    2017-11-01

    In this case study, metal pollution levels in mine wastes at a coal mine site in Etili coal mine (Can coal basin, NW Turkey) are evaluated using geographical information system (GIS) tools. Etili coal mine was operated since the 1980s as an open pit. Acid mine drainage is the main environmental problem around the coal mine. The main environmental contamination source is mine wastes stored around the mine site. Mine wastes were dumped over an extensive area along the riverbeds, and are now abandoned. Mine waste samples were homogenously taken at 10 locations within the sampling area of 102.33 ha. The paste pH and electrical conductivity values of mine wastes ranged from 2.87 to 4.17 and 432 to 2430 μS/cm, respectively. Maximum Al, Fe, Mn, Pb, Zn and Ni concentrations of wastes were measured as 109300, 70600, 309.86, 115.2, 38 and 5.3 mg/kg, respectively. The Al, Fe and Pb concentrations of mine wastes are higher than world surface rock average values. The geochemical analysis results from the study area were presented in the form of maps. The GIS based environmental database will serve as a reference study for our future work.

  3. Characterization of protein--DNA interactions using surface plasmon resonance spectroscopy with various assay schemes.

    PubMed

    Teh, Huey Fang; Peh, Wendy Y X; Su, Xiaodi; Thomsen, Jane S

    2007-02-27

    Specific protein-DNA interactions play a central role in transcription and other biological processes. A comprehensive characterization of protein-DNA interactions should include information about binding affinity, kinetics, sequence specificity, and binding stoichiometry. In this study, we have used surface plasmon resonance spectroscopy (SPR) to study the interactions between human estrogen receptors (ER, alpha and beta subtypes) and estrogen response elements (ERE), with four assay schemes. First, we determined the sequence-dependent receptors' binding capacity by monitoring the binding of ER to various ERE sequences immobilized on a sensor surface (assay format denoted as the direct assay). Second, we screened the relative affinity of ER for various ERE sequences using a competition assay, in which the receptors bind to an ERE-immobilized surface in the presence of competitor ERE sequences. Third, we monitored the assembly of ER-ERE complexes on a SPR surface and thereafter the removal and/or dissociation of the ER (assay scheme denoted as the dissociation assay) to determine the binding stoichiometry. Last, a sandwich assay (ER binding to ERE followed by anti-ER recognition of a specific ER subtype) was performed in an effort to understand how ERalpha and ERbeta may associate and compete when binding to the DNA. With these assay schemes, we reaffirmed that (1) ERalpha is more sensitive than ERbeta to base pair change(s) in the consensus ERE, (2) ERalpha and ERbeta form a heterodimer when they bind to the consensus ERE, and (3) the binding stoichiometry of both ERalpha- and ERbeta-ERE complexes is dependent on salt concentration. With this study, we demonstrate the versatility of the SPR analysis. With the involvement of various assay arrangements, the SPR analysis can be further extended to more than kinetics and affinity study.

  4. Flow cytometric immunobead assay for quantitative detection of platelet autoantibodies in immune thrombocytopenia patients.

    PubMed

    Zhai, Juping; Ding, Mengyuan; Yang, Tianjie; Zuo, Bin; Weng, Zhen; Zhao, Yunxiao; He, Jun; Wu, Qingyu; Ruan, Changgeng; He, Yang

    2017-10-23

    Platelet autoantibody detection is critical for immune thrombocytopenia (ITP) diagnosis and prognosis. Therefore, we aimed to establish a quantitative flow cytometric immunobead assay (FCIA) for ITP platelet autoantibodies evaluation. Capture microbeads coupled with anti-GPIX, -GPIb, -GPIIb, -GPIIIa and P-selectin antibodies were used to bind the platelet-bound autoantibodies complex generated from plasma samples of 250 ITP patients, 163 non-ITP patients and 243 healthy controls, a fluorescein isothiocyanate (FITC)-conjugated secondary antibody was the detector reagent and mean fluorescence intensity (MFI) signals were recorded by flow cytometry. Intra- and inter-assay variations of the quantitative FCIA assay were assessed. Comparisons of the specificity, sensitivity and accuracy between quantitative and qualitative FCIA or monoclonal antibody immobilization of platelet antigen (MAIPA) assay were performed. Finally, treatment process was monitored by our quantitative FCIA in 8 newly diagnosed ITPs. The coefficient of variations (CV) of the quantitative FCIA assay were respectively 9.4, 3.8, 5.4, 5.1 and 5.8% for anti-GPIX, -GPIb, -GPIIIa, -GPIIb and -P-selectin autoantibodies. Elevated levels of autoantibodies against platelet glycoproteins GPIX, GPIb, GPIIIa, GPIIb and P-selectin were detected by our quantitative FCIA in ITP patients compared to non-ITP patients or healthy controls. The sensitivity, specificity and accuracy of our quantitative assay were respectively 73.13, 81.98 and 78.65% when combining all 5 autoantibodies, while the sensitivity, specificity and accuracy of MAIPA assay were respectively 41.46, 90.41 and 72.81%. A quantitative FCIA assay was established. Reduced levels of platelet autoantibodies could be confirmed by our quantitative FCIA in ITP patients after corticosteroid treatment. Our quantitative assay is not only good for ITP diagnosis but also for ITP treatment monitoring.

  5. An alpha-synuclein MRM assay with diagnostic potential for Parkinson's disease and monitoring disease progression

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yang, Li; Stewart, Tessandra; Shi, Min

    Aim: The alpha-synuclein (α-syn) level in human cerebrospinal fluid (CSF), as measured by immunoassays, is promising as a Parkinson’s disease (PD) biomarker. However, the levels of total α-syn are inconsistent among studies with large cohorts and different measurement platforms. Total α-syn level also does not correlate with disease severity or progression. Here, we developed a highly sensitive Multiple Reaction Monitoring (MRM) method to measure absolute CSF α-syn peptide concentrations without prior enrichment or fractionation, aiming to discover new candidate biomarkers. Results: Six peptides covering 73% of protein sequence were reliably identified, and two were consistently quantified in cross-sectional and longitudinalmore » cohorts. Absolute concentration of α-syn in human CSF was determined to be 2.1ng/mL. A unique α-syn peptide, TVEGAGSIAAATGFVK (81-96), displayed excellent correlation with previous immunoassay results in two independent PD cohorts (p < 0.001), correlated with disease severity, and its changes significantly tracked the disease progression longitudinally. Conclusions: An MRM assay to quantify human CSF α-syn was developed and optimized. Sixty clinical samples from cross-sectional and longitudinal PD cohorts were analyzed with this approach. Although further larger-scale validation is needed, the results suggest that α-syn peptide could serve as a promising biomarker in PD diagnosis and progression.« less

  6. A fluorescence spectroscopy assay for real-time monitoring of enzyme immobilization into mesoporous silica particles.

    PubMed

    Nabavi Zadeh, Pegah S; Mallak, Kassam Abdel; Carlsson, Nils; Åkerman, Björn

    2015-05-01

    Mesoporous silica particles are used as support material for immobilization of enzymes. Here we investigated a fluorescence-based assay for real-time monitoring of the immobilization of lipase, bovine serum albumin, and glucose oxidase into micrometer-sized mesoporous silica particles. The proteins are labeled with the dye epicocconone, and the interaction with the particles is observed as an increase in emission intensity of the protein-dye conjugates that can be quantified if correcting for a comparatively slow photobleaching. The immobilization occurs in tens of minutes to hours depending on particle concentration and type of protein. In the limit of excess particles over proteins, the formation of the particle-protein complexes can be described by a single exponential growth for all three investigated proteins, and the fitted pseudo-first-order rate constant increases linearly with particle concentration for each protein type. The derived second-order rate constant k varies with the protein hydrodynamic radius according to k∼RH(-4.70±0.01), indicating that the rate-limiting step at high particle concentrations is not the diffusional encounter between proteins and particles but rather the entry into the pores, consistent with the hydrodynamic radii of the three proteins being smaller but comparable to the pore radius of the particles. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Biochemical methane potential (BMP) of agro-food wastes from the Cider Region (Spain).

    PubMed

    Nieto, P P; Hidalgo, D; Irusta, R; Kraut, D

    2012-01-01

    An inventory of agro-food industry organic waste streams with a high potential for biogas transformation was studied in a logistically viable area (Cider Region, Asturias, Spain). Three industries were selected as the most viable ones: livestock, dairy and beverage. The potential for methane production from six wastes (beverage waste, BW; milled apple waste, MA; milk waste, MK; yogurt waste, YG; fats and oils from dairy wastewater treatment, F&O and cattle manure, CM) at five different substrate:inoculum ratios (0.25, 0.50, 0.75, 1.00 and 1.50) was evaluated in laboratory batch assays. Obtained methane yields ranged from 202-549 mL STP CH(4)·g VS waste(-1), and the methane content in biogas ranged from 58-76%. The ultimate practical biochemical methane potentials were slightly affected by the substrate:inoculum ratio. The estimation of the regional fluxes of waste and methane potentials suggests anaerobic digestion as a sustainable solution for the valorization of the organic wastes generated in this Region.

  8. Targeted proteomic assays for quantitation of proteins identified by proteogenomic analysis of ovarian cancer

    DOE PAGES

    Song, Ehwang; Gao, Yuqian; Wu, Chaochao; ...

    2017-07-19

    Here, mass spectrometry (MS) based targeted proteomic methods such as selected reaction monitoring (SRM) are becoming the method of choice for preclinical verification of candidate protein biomarkers. The Clinical Proteomic Tumor Analysis Consortium (CPTAC) of the National Cancer Institute has investigated the standardization and analytical validation of the SRM assays and demonstrated robust analytical performance on different instruments across different laboratories. An Assay Portal has also been established by CPTAC to provide the research community a resource consisting of large set of targeted MS-based assays, and a depository to share assays publicly, providing that assays meet the guidelines proposed bymore » CPTAC. Herein, we report 98 SRM assays covering 70 candidate protein biomarkers previously reported as associated with ovarian cancer that have been thoroughly characterized according to the CPTAC Assay Characterization Guidance Document. The experiments, methods and results for characterizing these SRM assays for their MS response, repeatability, selectivity, stability, and reproducible detection of endogenous analytes are described in detail.« less

  9. Using Fish Tissue Data to Monitor Remedy Effectiveness

    EPA Science Inventory

    Chapter 8 of the Contaminated Sediment Remediation Guidance for Hazardous Waste Sites (OSWER Directive 9355.0-85, December 2005), presents an approach for developing an effective monitoring plan. As stated in the Guidance, one of the goals of monitoring is to “evaluate long-term ...

  10. RH-TRU Waste Inventory Characterization by AK and Proposed WIPP RH-TRU Waste Characterization Objectives

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Most, W. A.; Kehrman, R.; Gist, C.

    2002-02-26

    The U.S. Department of Energy (DOE)-Carlsbad Field Office (CBFO) has developed draft documentation to present the proposed Waste Isolation Pilot Plant (WIPP) remote-handled (RH-) transuranic (TRU) waste characterization program to its regulators, the U.S. Environmental Protection Agency and the New Mexico Environment Department. Compliance with Title 40, Code of Federal Regulations, Parts 191 and 194; the WIPP Land Withdrawal Act (PL 102-579); and the WIPP Hazardous Waste Facility Permit, as well as the Certificates of Compliance for the 72-B and 10-160B Casks, requires that specific waste parameter limits be imposed on DOE sites disposing of TRU waste at WIPP. Themore » DOE-CBFO must control the sites' compliance with the limits by specifying allowable characterization methods. As with the established WIPP contact handled TRU waste characterization program, the DOE-CBFO has proposed a Remote-Handled TRU Waste Acceptance Criteria (RH-WAC) document consolidating the requirements from various regulatory drivers and proposed allowable characterization methods. These criteria are consistent with the recommendation of a recent National Academy Sciences/National Research Council to develop an RH-TRU waste characterization approach that removes current self imposed requirements that lack a legal or safety basis. As proposed in the draft RH-WAC and other preliminary documents, the DOE-CBFO RH-TRU waste characterization program proposes the use of acceptable knowledge (AK) as the primary method for obtaining required characterization information. The use of AK involves applying knowledge of the waste in light of the materials or processes used to generate the waste. Documentation, records, or processes providing information about various attributes of a waste stream, such as chemical, physical, and radiological properties, may be used as AK and may be applied to individual waste containers either independently or in conjunction with radiography, visual examination, assay

  11. Monitoring the Secretory Behavior of the Rat Adrenal Medulla by High-Performance Liquid Chromatography-Based Catecholamine Assay from Slice Supernatants

    PubMed Central

    De Nardi, Frédéric; Lefort, Claudie; Bréard, Dimitri; Richomme, Pascal; Legros, Christian; Guérineau, Nathalie C.

    2017-01-01

    Catecholamine (CA) secretion from the adrenal medullary tissue is a key step of the adaptive response triggered by an organism to cope with stress. Whereas molecular and cellular secretory processes have been extensively studied at the single chromaffin cell level, data available for the whole gland level are much scarcer. We tackled this issue in rat by developing an easy to implement experimental strategy combining the adrenal acute slice supernatant collection with a high-performance liquid chromatography-based epinephrine and norepinephrine (NE) assay. This technique affords a convenient method for measuring basal and stimulated CA release from single acute slices, allowing thus to individually address the secretory function of the left and right glands. Our data point that the two glands are equally competent to secrete epinephrine and NE, exhibiting an equivalent epinephrine:NE ratio, both at rest and in response to a cholinergic stimulation. Nicotine is, however, more efficient than acetylcholine to evoke NE release. A pharmacological challenge with hexamethonium, an α3-containing nicotinic acetylcholine receptor antagonist, disclosed that epinephrine- and NE-secreting chromaffin cells distinctly expressed α3 nicotinic receptors, with a dominant contribution in NE cells. As such, beyond the novelty of CA assays from acute slice supernatants, our study contributes at refining the secretory behavior of the rat adrenal medullary tissue, and opens new perspectives for monitoring the release of other hormones and transmitters, especially those involved in the stress response. PMID:28993760

  12. Development of an atmospheric monitoring plan for space station

    NASA Technical Reports Server (NTRS)

    Casserly, Dennis M.

    1989-01-01

    An environmental health monitoring plan for Space Station will ensure crew health during prolonged habitation. The Space Station, Freedom, will operate for extended periods, 90+ days, without resupply. A regenerative, closed loop life support system will be utilized in order to minimize resupply logistics and costs. Overboard disposal of wastes and venting of gases to space will be minimal. All waste material will be treated and recycled. The concentrated wastes will be stabilized and stored for ground disposal. The expected useful life of the station (decades) and the diversity of materials brought aboard for experimental or manufacturing purposes, increases the likelihood of cabin contamination. Processes by which cabin contamination can occur include: biological waste production, material off-gassing, process leakage, accidental containment breach, and accumulation due to poor removal efficiencies of the purification units. An industrial hygiene approach was taken to rationalize monitoring needs and to identify the substances likely to be present, the amount, and their hazard.

  13. Impact of electronic waste disposal on lead concentrations in landfill leachate.

    PubMed

    Spalvins, Erik; Dubey, Brajesh; Townsend, Timothy

    2008-10-01

    Lead is the element most likely to cause discarded electronic devices to be characterized as hazardous waste. To examine the fate of lead from discarded electronics in landfills, five columns were filled with synthetic municipal solid waste (MSW). A mix of electronic devices was added to three columns (6% by weight), while two columns served as controls. A sixth column contained waste excavated from an existing MSW landfill. Leachate quality was monitored for 440 days. In columns with the synthetic waste, leachate pH indicated that the simulated landfill environment was characteristic of the acid phase of waste decomposition; lead leachability should be greater in the acid phase of landfill degradation as compared to the methanogenic phase. Lead concentrations ranged from 7 to 66 microg/L in the columns containing electronic waste and ranged from < 2 to 54 microg/L in the control columns. Although the mean lead concentrations in the columns containing electronic devices were greater than those in the controls, the difference was not found to be statistically significant when comparing the data sets over the entire monitoring period. Lead results from the excavated waste column suggest that lead concentrations in all columns will decrease as the pH increases toward more neutral methanogenic conditions.

  14. Comparison of balance of tritium activity in waste water from nuclear power plants and at selected monitoring sites in the Vltava River, Elbe River and Jihlava (Dyje) River catchments in the Czech Republic.

    PubMed

    Hanslík, Eduard; Marešová, Diana; Juranová, Eva; Sedlářová, Barbora

    2017-12-01

    During the routine operation, nuclear power plants discharge waste water containing a certain amount of radioactivity, whose main component is the artificial radionuclide tritium. The amounts of tritium released into the environment are kept within the legal requirements, which minimize the noxious effects of radioactivity, but the activity concentration is well measurable in surface water of the recipient. This study compares amount of tritium activity in waste water from nuclear power plants and the tritium activity detected at selected relevant sites of surface water quality monitoring. The situation is assessed in the catchment of the Vltava and Elbe Rivers, affected by the Temelín Nuclear Power Plant as well as in the Jihlava River catchment (the Danube River catchment respectively), where the waste water of the Dukovany Nuclear Power Plant is discharged. The results show a good agreement of the amount of released tritium stated by the power plant operator and the tritium amount detected in the surface water and highlighted the importance of a robust independent monitoring of tritium discharged from a nuclear power plant which could be carried out by water management authorities. The outputs of independent monitoring allow validating the values reported by a polluter and expand opportunities of using tritium as e.g. tracer. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Comparison of the Abbott RealTime HCV and Roche COBAS Ampliprep/COBAS TaqMan HCV assays for the monitoring of sofosbuvir-based therapy.

    PubMed

    Ogawa, Eiichi; Furusyo, Norihiro; Murata, Masayuki; Shimizu, Motohiro; Toyoda, Kazuhiro; Hotta, Taeko; Uchiumi, Takeshi; Hayashi, Jun

    2017-01-01

    On-treatment HCV kinetics play an invaluable role in evaluating the efficacy of interferon-based therapies. However, the importance of HCV RNA monitoring has not been well discussed concerning treatment with sofosbuvir (SOF)-based regimens, especially for the utility of the Abbott RealTime HCV (ART) assay. This study consisted of 151 patients infected with HCV genotype-1 or -2, including patients with prior treatment-experience or cirrhosis. HCV genotype-1 patients were treated with SOF/ledipasvir and genotype-2 patients with SOF/ribavirin, both for 12 weeks. Serial measurements of HCV RNA were performed with both the ART and COBAS AmpliPrep/COBAS TaqMan v2.0 (CAP/CTM) assays simultaneously at weeks 0, 1, 2, 4, 6, 8, 10 and 12 of treatment. The rates of HCV RNA target not detected (TND) by ART were significantly lower than those by CAP/CTM between weeks 2 and 12 (end of treatment [EOT]), irrespective of prior treatment-experience or cirrhosis. 11 (11.6%) genotype-1 and 8 (14.3%) genotype-2 patients did not achieve HCV RNA TND by ART at EOT, in contrast to all having HCV RNA TND by CAP/CTM; however, all achieved sustained virological response. The time at which HCV RNA became TND or unquantifiable was not associated with treatment outcome by either the ART or CAP/CTM assay. Over 10% of the patients continued to have detectable HCV RNA by ART at EOT, irrespective of HCV genotype, prior treatment-experience and/or cirrhosis. However, prolonged residual HCV RNA was not associated with treatment failure.

  16. Eco-efficient waste glass recycling: Integrated waste management and green product development through LCA.

    PubMed

    Blengini, Gian Andrea; Busto, Mirko; Fantoni, Moris; Fino, Debora

    2012-05-01

    As part of the EU Life + NOVEDI project, a new eco-efficient recycling route has been implemented to maximise resources and energy recovery from post-consumer waste glass, through integrated waste management and industrial production. Life cycle assessment (LCA) has been used to identify engineering solutions to sustainability during the development of green building products. The new process and the related LCA are framed within a meaningful case of industrial symbiosis, where multiple waste streams are utilised in a multi-output industrial process. The input is a mix of rejected waste glass from conventional container glass recycling and waste special glass such as monitor glass, bulbs and glass fibres. The green building product is a recycled foam glass (RFG) to be used in high efficiency thermally insulating and lightweight concrete. The environmental gains have been contrasted against induced impacts and improvements have been proposed. Recovered co-products, such as glass fragments/powders, plastics and metals, correspond to environmental gains that are higher than those related to landfill avoidance, whereas the latter is cancelled due to increased transportation distances. In accordance to an eco-efficiency principle, it has been highlighted that recourse to highly energy intensive recycling should be limited to waste that cannot be closed-loop recycled. Copyright © 2011 Elsevier Ltd. All rights reserved.

  17. Characterization, monitoring, and sensor technology catalogue

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Matalucci, R.V.; Esparza-Baca, C.; Jimenez, R.D.

    1995-12-01

    This document represents a summary of 58 technologies that are being developed by the Department of Energy`s (DOE`s) Office of Science and Technology (OST) to provide site, waste, and process characterization and monitoring solutions to the DOE weapons complex. The information was compiled to provide performance data on OST-developed technologies to scientists and engineers responsible for preparing Remedial Investigation/Feasibility Studies (RI/FSs) and preparing plans and compliance documents for DOE cleanup and waste management programs. The information may also be used to identify opportunities for partnering and commercialization with industry, DOE laboratories, other federal and state agencies, and the academic community.more » Each technology is featured in a format that provides: (1) a description, (2) technical performance data, (3) applicability, (4) development status, (5) regulatory considerations, (6) potential commercial applications, (7) intellectual property, and (8) points-of-contact. Technologies are categorized into the following areas: (1) Bioremediation Monitoring, (2) Decontamination and Decommissioning, (3) Field Analytical Laboratories, (4) Geophysical and Hydrologic Characterization, (5) Hazardous Inorganic Contaminant Analysis, (6) Hazardous Organic Contaminant Analysis, (7) Mixed Waste, (8) Radioactive Contaminant Analysis, (9) Remote Sensing,(10)Sampling and Drilling, (11) Statistically Guided Sampling, and (12) Tank Waste.« less

  18. Calendar Year 2016 Annual Groundwater Monitoring Report.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Copland, John R.; Jackson, Timmie Okchumpulla; Li, Jun

    Sandia National Laboratories, New Mexico (SNL/NM) is a government-owned/contractoroperated laboratory. National Technology and Engineering Solutions of Sandia, LLC, a wholly owned subsidiary of Honeywell International, Inc., manages and operates SNL/NM for the U.S. Department of Energy (DOE), National Nuclear Security Administration (NNSA). The DOE/NNSA Sandia Field Office administers the contract and oversees contractor operations at the site. Two types of groundwater surveillance monitoring are conducted at SNL/NM: (1) on a site-wide basis as part of the SNL/NM Long-Term Stewardship (LTS) Program’s Groundwater Monitoring Program (GMP) Groundwater Surveillance Task and (2) on a site-specific groundwater monitoring at LTS/Environmental Restoration (ER) Operationsmore » sites with ongoing groundwater investigations. This Annual Groundwater Monitoring Report summarizes data collected during groundwater monitoring events conducted at GMP locations and at the following SNL/NM sites through December 31, 2016: Burn Site Groundwater Area of Concern (AOC); Chemical Waste Landfill; Mixed Waste Landfill; Technical Area-V Groundwater AOC; and the Tijeras Arroyo Groundwater AOC. Environmental monitoring and surveillance programs are required by the New Mexico Environment Department (NMED) and DOE Order 436.1, Departmental Sustainability, and DOE Order 231.1B, Environment, Safety, and Health Reporting.« less

  19. Chromogenic assay for BAY 81-8973 potency assignment has no impact on clinical outcome or monitoring in patient samples.

    PubMed

    Kitchen, S; Katterle, Y; Beckmann, H; Maas Enriquez, M

    2016-06-01

    Essentials Discrepancies can exist in factor VIII activity measured by the one-stage or chromogenic assays. LEOPOLD trial data were used to assess clinical impact of BAY 81-8973 potency assignment assay. Efficacy was not affected by the assay used for potency assignment and dosing of BAY 81-8973. Either assay may be used to measure factor VIII activity after BAY 81-8973 infusion. Background Product-specific discrepancies have been reported for factor VIII (FVIII) activity determined with one-stage or chromogenic assays. Objective To assess the clinical impact of potency assignment of BAY 81-8973, a full-length, unmodified, recombinant human FVIII, by use of the chromogenic assay or chromogenic assay adjusted to mimic results obtained with the one-stage assay Patients/methods Patients aged 12-65 years with severe hemophilia A received BAY 81-8973 in LEOPOLD I (20-50 IU kg(-1) two or three times weekly [investigator decision]) and LEOPOLD II (randomized to 20-30 IU kg(-1) twice weekly, 30-40 IU kg(-1) three times weekly, or on-demand treatment). Both trials included two 6-month crossover periods in which potency labeling was determined with the chromogenic substrate assay as per the European Pharmacopoeia (CS/EP) or the chromogenic substrate assay adjusted to mimic results obtained with the one-stage assay (CS/ADJ). The annualized bleeding rate (ABR) and FVIII incremental recovery were assessed by the use of pooled data. Results The analysis was perfomed on 121 patients. Median (quartile [Q] 1; Q3) ABRs during the CS/EP and CS/ADJ periods were 1.98 (0; 5.92) and 1.98 (0; 7.34), respectively. The mean incremental recovery was > 2 IU dL(-1) per IU kg(-1) in both periods with the use of either assay for postinfusion FVIII measurements. The median (Q1; Q3) chromogenic/one-stage assay recovery ratio was 1.054 (0.892; 1.150) for the CS/EP period when a plasma standard was used for calibration. Conclusions No impact on the ABR was observed with chromogenic-based as compared

  20. Controls for Burning Solid Wastes

    ERIC Educational Resources Information Center

    Toro, Richard F.; Weinstein, Norman J.

    1975-01-01

    Modern thermal solid waste processing systems are becoming more complex, incorporating features that require instrumentation and control systems to a degree greater than that previously required just for proper combustion control. With the advent of complex, sophisticated, thermal processing systems, TV monitoring and computer control should…