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A bead-based method for multiplexed identification and quantitation of DNA sequences using flow cytometry.

A new multiplexed, bead-based method which utilizes nucleic acid hybridizations on the surface of microscopic polystyrene spheres to identify specific sequences in heterogeneous mixtures of DNA sequences is described. The method consists of three elements: beads (5.6-microm diameter) with oligomer capture probes attached to the surface, three fluorophores for multiplexed detection, and flow cytometry instrumentation. Two fluorophores are impregnated within each bead in varying amounts to create different bead types, each associated with a unique probe. The third fluorophore is a reporter. Following capture of fluorescent cDNA sequences from environmental samples, the beads are analyzed by flow cytometric techniques which yield a signal intensity for each capture probe proportional to the amount of target sequences in the analyte. In this study, a direct hybrid capture assay was developed and evaluated with regard to sequence discrimination and quantitation of abundances. The target sequences (628 to 728 bp in length) were obtained from the 16S/23S intergenic spacer region of microorganisms collected from polluted groundwater at the nuclear waste site in Hanford, Wash. A fluorescence standard consisting of beads with a known number of fluorescent DNA molecules on the surface was developed, and the resolution, sensitivity, and lower detection limit for measuring abundances were determined. The results were compared with those of a DNA microarray using the same sequences. The bead method exhibited far superior sequence discrimination and possesses features which facilitate accurate quantitation.

Base Sequence↗

Tritium release from nuclear power plants in Taiwan.

Tritium is produced in light water reactors from various sources and the design source terms for both BWR and PWR plants are reviewed. The chemical forms of tritium produced in the coolant are discussed in terms of water radiolysis and free radical reactions in the reactor core regions, and the major form of tritium released from the coolant systems is confirmed to be tritiated water. The tritium activity concentrations and inventories in various coolant systems have been measured, and the release pathways of tritium from nuclear power plants are also reviewed in this paper. Decreasing trends of tritium release from nuclear power plants in both liquid waste and ventilation sources have been observed in Taiwan. The impact of tritium release on environmental radiation is estimated with well-established screening models, and the results confirm that the impact is less than 1% of the regulatory limits and less than 0.1% of the natural radiation background.

Air Pollutants, Radioactive↗

Revision of fee schedules; 100% fee recovery, FY 1992--NRC. Final rule.

The Nuclear Regulatory Commission (NRC) is amending the licensing, inspection, and annual fees charged to its applicants and licensees. The amendments are necessary to implement Public Law 101-508, signed into law on November 5, 1990, which mandates that the NRC recover approximately 100 percent of its budget authority in Fiscal Year (FY) 1992 less amounts appropriated from the Nuclear Waste Fund (NWF). The amount to be recovered for FY 1992 is approximately $492.5 million.

Fees and Charges↗

Changes in uranium speciation through a depth sequence of contaminated Hanford sediments.

The disposal of basic sodium aluminate and acidic U(VI)-Cu(ll) wastes in the now-dry North and South 300 A Process Ponds atthe Hanford site resulted in a groundwater plume of U(VI). To gain insight into the geochemical processes that occurred during waste disposal and those affecting the current and future fate and transport of this uranium plume, the solid-phase speciation of uranium in a depth sequence of sediments from the base of the North Process Pond through the vadose zone to groundwater was investigated using standard chemical and mineralogical analyses, electron and X-ray microprobe measurements, and X-ray absorption fine structure spectroscopy. Near-surface sediments contained uranium coprecipitated with calcite, which formed due to overneutralization of the waste ponds with base (NaOH). At intermediate depths in the vadose zone, metatorbernite [Cu(UO2PO4)2 x 8H2O] precipitated, likely during pond operations. Uranium occurred predominantly sorbed onto phyllosilicates in the deeper vadose zone and groundwater; sorbed uranium was also an important component at intermediate depths. Since the calcite-bearing pond sediments have been removed in remediation efforts, uranium fate and transport will be controlled primarily by desorption of the sorbed uranium and dissolution of metatorbernite.

Adsorption↗

Revision and meta-analysis of selected biosphere parameter values for chlorine, iodine, neptunium, radium, radon and uranium.

There is a continual supply of new experimental data that are relevant to the assessment of the potential impacts of nuclear fuel waste disposal. In the biosphere, the traditional assessment models are data intensive, and values are needed for several thousand parameters. This is augmented further when measures of central tendency, statistical dispersion, correlations and truncations are required for each parameter to allow probabilistic risk assessment. Recent reviews proposed values for 10-15 key element-specific parameters relevant to (36)Cl, (129)I, (222)Rn, (226)Ra, (237)Np and (238)U, and some highlights from this data update are summarized here. Several parameters for Np are revised downward by more than 10-fold, as is the fish/water concentration ratio for U. Soil solid/liquid partition coefficients, Kd, are revised downward by 10-770-fold for Ra. Specific parameters are discussed in detail, including degassing of I from soil; sorption of Cl in soil; categorization of plant/soil concentration ratios for U, Ra and Np; Rn transfer from soil to indoor air; Rn degassing from surface water; and the Ca dependence of Ra transfers.

Animals↗

Laser ablation inductively coupled plasma mass spectrometry measurement of isotope ratios in depleted uranium contaminated soils.

Laser ablation of pressed soil pellets was examined as a means of direct sample introduction to enable inductively coupled plasma mass spectrometry (ICP-MS) screening of soils for residual depleted uranium (DU) contamination. Differentiation between depleted uranium, an anthropogenic contaminant, and naturally occurring uranium was accomplished on the basis of measured 235U/238U isotope ratios. The amount of sample preparation required for laser ablation is considerably less than that typically required for aqueous sample introduction. The amount of hazardous laboratory waste generated is diminished accordingly. During the present investigation, 235U/238U isotope ratios measured for field samples were in good agreement with those derived from gamma spectrometry measurements. However, substantial compensation was required to mitigate the effects of impaired pulse counting attributed to sample inhomogeneity and sporadic introduction of uranium analyte into the plasma.

Background Radiation↗

Health physics challenges involved with opening a "seventeen-inch" concrete waste vault.

This paper describes the various activities involved with opening a sealed legacy "Seventeen-inch" concrete vault and the health physics challenges and solutions employed. As part of a legacy waste stream that was removed from the former Hazardous Waste Management Facility at Brookhaven National Laboratory, the "Seventeen-inch" concrete vault labeled 1-95 was moved to the new Waste Management Facility for ultimate disposal. Because the vault contained 239Pu foils with a total activity in excess of the transuranic waste limits, the foils needed to be removed and repackaged for disposal. Conventional diamond wire saws could not be used because of facility constraints, so this project relied mainly on manual techniques. The planning and engineering controls put in place enabled personnel to open the vault and remove the waste while keeping dose as low as reasonably achievable.

Decontamination↗

Transfer of 137Cs and 60Co in a waste retention pond with emphasis on aquatic insects.

The objectives of this research were (1) to analyze the transfers of 137Cs and 60Co in a retention pond, with emphasis on aquatic insects and (2) to determine if detectable concentrations of these radionuclides are exported by emerging aquatic insects. We analyzed the radionuclide concentrations in the following components: water solution, bottom sediments, suspended particulate matter, plankton, floating mats of filamentous algae, benthic macroinvertebrates, and emerging aquatic insects. Samples were collected quarterly from June 1981 to April 1982. The lowest concentrations (in picocuries per milliliter) occurred in solution (range: 1.4 X 10(2) to 3.2 X 10(2) for 137Cs and 8.1 X 10(-1) to 2.2 X 10(0) for 60Co). The highest concentrations (in picocuries per gram dry weight) occurred in the sediments (range: 1.5 X 10(4) to 1.1 X 10(8) for 137Cs and 1.0 X 10(2) to 4.3 X 10(6) for 60Co). The primary producers and aquatic insect consumers had concentrations of both radionuclides that were two to four orders of magnitude higher than the respective concentrations dissolved in water but two to three orders of magnitude lower than the concentrations in the sediments. The concentrations of both radionuclides decreased successively at higher trophic levels. There were considerable temporal variations as the radionuclides cycled among the abiotic and biotic components of the pond. Emerging adult aquatic insects had lower concentrations of both radionuclides than the immature stages that lived in the pond (adult/immature ratio about 0.25). Because the emerging adult insects contain detectable concentrations of radionuclides, have relatively long life spans, and disperse away from the aquatic habitat, we conclude that adult aquatic insects would be effective biological monitors.

Animals↗

[Automatic collection system of waste scintillator solution using micro robots].

Industrial micro-robots are now being applied to hazardous jobs or tedious jobs. These in-house micro-robots (MELFA, RM-501, Mitsubishi Electric Inc.) were used to make a collection system of waste scintillator solution on the experimental basis. Newly developed were hands, vial holders and a system soft ware, and satisfactory results have been obtained.

Automation↗

Toxic agent and radiation control: progress toward objectives for the nation for the year 1990.

In 1980, the Department of Health and Human Services set national prevention objectives for 1990 in 15 health priority areas, 1 of which is the control of toxic agents and radiation. Ten objectives related to this area are priorities for the national control effort. Progress is reviewed on those priorities within the responsibilities of the Public Health Service. Six key program elements, or types of support activities, are deemed essential to preventing, identifying, and controlling toxic agent and radiation threats. Significant progress has been made toward achieving objectives for which all key program elements have been successfully implemented to provide the requisite know-how, manpower, and tools. Important advances have been made in reducing the blood lead levels of the population, reducing unnecessary exposure to medical X-rays, evaluating the toxicities of chemicals in toxic waste dumps, and improving the scientific and technical information base and its availability for prevention and control efforts. The most important priority for the forseeable future will be to expand our knowledge of potential health risks posed by toxic agents and radiation. Expanded surveillance systems and data bases are essential to determining the extent of the problems in terms of human health effects and for measuring the impact of prevention programs. Emphasis on the activities embodied in the key elements will encourage the expansion of the knowledge base and its effective application to prevention and control problems.

Environmental Health↗

Development of a chemical form discrimination system for 14C species in gaseous waste.

A new system was developed for discriminating the chemical form of 14C species in gaseous waste from nuclear facilities. The system consists of an air sample reservoir, a concentration apparatus, a radio-gas chromatograph, and microcomputers. An air sample, which is part of the gaseous effluent released from a stack of a facility, is automatically collected in the tank by trigger signals of stack monitors. After passing the air through a silica gel column, carbon species contained in it are recovered in an activated carbon-supported palladium trap held at -120 degrees C. The recovered species are swept with helium gas and analyzed by the radiogas chromatograph using an activated carbon column and gas-flow counters. Output signals of the gas-flow counters are continuously recorded by the microcomputer. By using this system, the chemical forms of 14C, such as CO, CO2, CH4, C2H4, C2H6, and C3H8 are identified and quantified. A detectable concentration of 0.3 mBq cm-3 for 14CO2 can be obtained. The system has been used to monitor the gaseous waste released from the radioisotope production laboratory and the Japan Research Reactor 2 at Japan Atomic Energy Research Institute.

Air Pollutants, Radioactive↗

Handling, storage, treatment, and disposal of mixed wastes at medical facilities and academic institutions.

Mixed waste includes hazardous/ toxic chemicals and radioactive materials mixed together. Most mixed wastes are difficult and costly to work with safely. Meeting the legal and regulatory requirements to generate, use, store, treat, and dispose of mixed waste can also be costly and difficult. The regulations that govern mixed waste, from creation to final disposition, are extensive and complex to implement. Complete and documented compliance with the applicable regulations is essential to demonstrate that personnel and environment safety has been assured. Noncompliance with the multitude of government's requirements for mixed waste can lead to serious repercussions. Noncompliance may cause serious harm to individuals and the environment, fines and penalties against a facility, and/or suspension of a facility's operations. Each facility that generates, uses, or possesses mixed waste must employ adequate expertise to assure full regulatory compliance. This paper introduces the large scope and variety of mixed waste and the current and potential methods to treat and ultimately dispose of, or recycle, mixed waste. Alternatives for regulatory compliance are also discussed and effective means to control mixed waste are presented. The paper is based on my Master's Thesis (Hageman 2001).

Academic Medical Centers↗

TRU waste absorbent addition project at the Idaho National Engineering and Environmental Laboratory.

ABSTRACT In order to meet a commitment to ship 3,100 m3 of transuranic waste to the Waste Isolation Pilot Plant (WIPP), the Idaho National Engineering and Environmental Laboratory (INEEL) developed a process to add absorbent to TRU waste drums that did not meet WIPP waste acceptance criteria. The development, implementation, and safe completion of this project contributed to the INEEL's success in meeting the commitment three months early.

Idaho↗

Pilot-scale in situ bioremediation of uranium in a highly contaminated aquifer. 1. Conditioning of a treatment zone.

To evaluate the potential for in situ bioremediation of U(VI) to sparingly soluble U(IV), we constructed a pilot test facility at Area 3 of the U.S. Department of Energy Natural and Accelerated Bioremediation Research (NABIR) Field Research Center (FRC) in Oak Ridge, TN. The facility is adjacent to the former S-3 Ponds which received trillions of liters of acidic plating wastes. High levels of uranium are present, with up to 800 mg kg(-1) in the soil and 84-210 microM in the groundwater. Ambient groundwater has a highly buffered pH of approximately 3.4 and high levels of aluminum (12-13 mM), calcium (22-25 mM), and nitrate (80-160 mM). Adjusting the pH of groundwater to approximately 5 within the aquifer would deposit extensive aluminum hydroxide precipitate. Calcium is present in the groundwater at levels that inhibit U(VI) reduction, but its removal by injection of a high pH solution would generate clogging precipitate. Nitrate also inhibits U(VI) reduction and is present at such high concentrations that its removal by in situ denitrification would generate large amounts of N2 gas and biomass. To establish and maintain hydraulic control, we installed a four well recirculation system parallel to geologic strike, with an inner loop nested within an outer loop. For monitoring, we drilled three boreholes perpendicular to strike across the inner loop and installed multilevel sampling tubes within them. A tracer pulse with clean water established travel times and connectivity between wells and enabled the assessment of contaminant release from the soil matrix. Subsequently, a highly conductive region of the subsurface was prepared for biostimulation by removing clogging agents and inhibitors and increasing pH. For 2 months, groundwater was pumped from the hydraulically conductive zone; treated to remove aluminum, calcium, and nitrate, and supplemented with tap water; adjusted to pH 4.3-4.5; then returned to the hydraulically conductive zone. This protocol removed most of the aqueous aluminum and calcium. The pH of the injected treated water was then increased to 6.0-6.3. With additional flushing, the pH of the extracted water gradually increased to 5.5-6.0, and nitrate concentrations fell to 0.5-1.0 mM. These conditions were judged suitable for biostimulation. In a companion paper (Wu et al., Environ. Sci. Technol. 2006, 40, 3978-3987), we describe the effects of ethanol addition on in situ denitrification and U(VI) reduction and immobilization.

Bacteria↗

Germination of seeds from an irradiated forest: implications for waste disposal.

Jack pine (Pinus banksiana Lamb.) retain their seeds from year to year, so that in an irradiated forest, each tree receives a specific dose rate but has seeds that have accumulated a range of total doses. The Field Irradiator-Gamma facility in Pinawa, Manitoba, contains jack pine that have been irradiated longer and at lower dose rates than previously reported. Seed germination and germination rate were examined on seeds irradiated on the parent tree for up to 5 years. Germination rate was most sensitive and showed deleterious effects at 1.1 mGy hr-1. This is not much lower than results reported by others in shorter-term studies. Effects were related to dose rate rather than total dose. Hormesis, indicated by statistically significant increased germination rate, was evident at 0.6 mGy hr-1. To put these results into context, the concentrations of selected radionuclides that, through internal contamination, would deliver 1.1 mGy hr-1 to plants were estimated. For 99Tc and 129I, these concentrations are far above the chemical toxicity thresholds for plants. Clearly, as assessments of waste repositories begin to consider effects on organisms other than humans, such as plants, chemical toxicity will be an important feature.

Dose-Response Relationship, Radiation↗

[The treatment of thyroid diseases using 131I in a special ward with a liquid waste storage plant].

This paper deals with the calculation of doses for organs of special interest under 131I treatment of thyroid diseases. Also the effective dose equivalent to the body of the patient, exclusive of the thyroid, is given. The ratio between the exposure by self-irradiation and the exposure from another patient in 2 m distance both having undergone 131I treatment with 50% uptake, is about 1000:1. Therefore positioning of a lead shield between two such patients in the same room is, from the point of view of radiation protection, unnecessary. A complete mathematical description of iodine distribution in a nuclear medicine ward and its liquid waste storage plant is given. The use in the authors' own department of these equations is demonstrated. Computer programs for additional calculations are available.

Humans↗

Variability in background levels of surface soil radionuclides in the vicinity of the US DOE waste isolation pilot plant.

Concentrations of radionuclides were measured in soils from a grid of locations surrounding the US Department of Energy Waste Isolation Pilot Plant (WIPP) in southeastern New Mexico and from a grid on a reference site approximately 20 km southeast of the WIPP site. Each of the two grids has 16 sampling locations (grid nodes) systematically distributed within an area of 16.580 ha. Sampling was conducted prior to the arrival of the first waste shipment at WIPP. Thus, the 137Cs and 23,240Pu in the soil are expected to have been deposited as global fallout, although the Gnome Site, 8.8 km southwest of the WIPP, is also a potential source of 239,240Pu and fission products. The reference grid has significantly higher concentrations of fallout and natural radionuclides than the WIPP grid. Up to 80% of the total variability in radionuclide concentrations across the two grids is attributable to differences between grid nodes. Differences between replicates within a location account for 44-50% of the variability in concentrations of the uranium isotopes, but only 11-17% of the variability in the concentrations of the other radionuclides. Samples having similar abundance of radionuclides were spatially aggregated across the terrain. The activity concentrations of the radionuclides were strongly correlated with the concentrations of Al and Pb, and with the percentages of sand, silt and clay in the soil. Normalizing radionuclide concentrations to the concentration of Al or percent fine particles can help adjust for differences in soil textures among samples and facilitate the detection of gradients or temporal changes in soil concentrations.

Aluminum↗