Characterization of metals released from coal fly ash during dredging at the Kingston ash recovery project.

A storage-pond dike failure occurred on December 22, 2008 at the Tennessee Valley Authority Kingston Fossil Plant resulting in the release of over 4million cubic meters (5million cubic yards) of fly ash. Approximately half of the released ash was deposited in the main channel of the Emory River, Tennessee, USA. Remediation efforts of the Emory River focused on hydraulic dredging, as well as mechanical excavation in targeted areas. However, agitation of the submerged fly ash during hydraulic dredging introduces river water into the fly ash material, which could promote dissolution and desorption of metals from the solid fly ash material. Furthermore, aeration of the dredge slurry could alter the redox state of metals in the fly ash material and thereby change their sorption, mobility, and toxicity properties. The research presented here focuses on the concentrations and speciation of metals during the fly ash recovery from the Emory River. Our results indicate that arsenite [As(III)] released from the fly ash material during dredging was slowly oxidized to arsenate [As(V)] in the slurry recovery system with subsequent removal through precipitation or sorption reactions with suspended fly ash material. Concentrations of other dissolved metals, including iron and manganese, also generally decreased in the ash recovery system prior to water discharge back to the river.

Growth rates are related to production efficiencies in juveniles of the sea urchin Lytechinus variegatus.

Growth rates of newly-metamorphosed urchins from a single spawning event (three males and three females) were highly variable, despite being held en masse under identical environmental and nutritional conditions. As individuals reached ~5 mm diameter (0.07-0.10 g wet weight), they were placed in growth trials (23 dietary treatments containing various nutrient profiles). Elapsed time from the first individual entering the growth trials to the last individual entering was 121 days (N = 170 individuals). During the five-week growth trials, urchins were held individually and proffered a limiting ration to evaluate growth rate and production efficiency. Growth rates among individuals within each dietary treatment remained highly variable. Across all dietary treatments, individuals with an initially high growth rate (entering the study first) continued to grow at a faster rate than those with an initially low growth rate (entering the study at a later date), regardless of feed intake. Wet weight gain (ranging from 0.13 -3.19 g, P < 0.0001, R2 = 0.5801) and dry matter production efficiency (ranging from 25.2-180.5%, P = 0.0003, R2 = 0.6162) were negatively correlated with stocking date, regardless of dietary treatment. Although canalization of growth rate during en masse early post-metamorphic growth is possible, we hypothesize that intrinsic differences in growth rates are, in part, the result of differences (possibly genetic) in production efficiencies of individual Lytechinus variegatus. That is, some sea urchins are more efficient in converting feed to biomass. We further hypothesize that this variation may have evolved as an adaptive response to selective pressure related to food availability.

Analysis of munitions constituents in groundwater using a field-portable GC-MS.

The use of munitions constituents (MCs) at military installations can produce soil and groundwater contamination that requires periodic monitoring even after training or manufacturing activities have ceased. Traditional groundwater monitoring methods require large volumes of aqueous samples (e.g., 2-4 L) to be shipped under chain of custody, to fixed laboratories for analysis. The samples must also be packed on ice and shielded from light to minimize degradation that may occur during transport and storage. The laboratory's turn-around time for sample analysis and reporting can be as long as 45 d. This process hinders the reporting of data to customers in a timely manner; yields data that are not necessarily representative of current site conditions owing to the lag time between sample collection and reporting; and incurs significant shipping costs for samples. The current work compares a field portable Gas Chromatograph-Mass Spectrometer (GC-MS) for analysis of MCs on-site with traditional laboratory-based analysis using High Performance Liquid Chromatography with UV absorption detection. The field method provides near real-time (within ~1 h of sampling) concentrations of MCs in groundwater samples. Mass spectrometry provides reliable confirmation of MCs and a means to identify unknown compounds that are potential false positives for methods with UV and other non-selective detectors.

Geochemical investigations of metals release from submerged coal fly ash using extended elutriate tests.

A storage pond dike failure occurred at the Tennessee Valley Authority Kingston Fossil Plant that resulted in the release of over 3.8 million cubic meters (5 million cubic yards) of fly ash. Approximately half of this material deposited in the main channel of the Emory River, 3.5 km upstream of the confluence of the Emory and Clinch Rivers, Tennessee, USA. Remediation efforts to date have focused on targeted removal of material from the channel through hydraulic dredging, as well as mechanical excavation in some areas. The agitation of the submerged fly ash during hydraulic dredging introduces river water into the fly ash material, which could alter the redox state of metals present in the fly ash and thereby change their sorption and mobility properties. A series of extended elutriate tests were used to determine the concentration and speciation of metals released from fly ash. Results indicated that arsenic and selenium species released from the fly ash materials during elutriate preparation were redox stable over the course of 10d, with dissolved arsenic being present as arsenate, and dissolved selenium being present as selenite. Concentrations of certain metals, such as arsenic, selenium, vanadium, and barium, increased in the elutriate waters over the 10d study, whereas manganese concentrations decreased, likely due to oxidation and precipitation reactions.

A modified acid digestion procedure for extraction of tungsten from soil.

Interest in tungsten occurrence and geochemistry is increasing due to increased use of tungsten compounds and its unknown biochemical effects. Tungsten has a complex geochemistry, existing in most environmental matrices as the soluble and mobile tungstate anion, as well as poly- and heteropolytungstates. Because the geochemistry of tungsten is substantially different than most trace metals, including the formation of insoluble species under acidic conditions, it is not extracted from soil matrices using standard acid digestion procedures. Therefore, the current work describes a modification to a commonly used acid digestion procedure to facilitate quantification of tungsten in soil matrices. Traditional soil digestion procedures, using nitric and hydrochloric acids with hydrogen peroxide yield <1 up to 50% recovery on soil matrix spike samples, whereas the modified method reported here, which includes the addition of phosphoric acid, yields spike recoveries in the 76-98% range. Comparison of the standard and modified digestion procedures on National Institute of Standards and Technology Standard Reference Materials yielded significantly improved tungsten recoveries for the phosphoric acid modified method. The modified method also produces comparable results for other acid extractable metals as the standard methods, and therefore can be used simultaneously for tungsten and other metals of interest.

Investigations of tungsten mobility in soil using column tests.

The geochemistry of tungsten has recently gained attention in the scientific and regulatory communities. Tungsten has a complex geochemistry, existing in many environmental matrices as the soluble and mobile tungstate anion, as well as a series of ill-defined polymeric species. Previous work has shown that soluble tungsten leached from a metallic tungsten-spiked Grenada Loring soil will reach an equilibrium concentration >150 mgL(-1), and the concentration is greatly influenced by co-occurring analytes in the matrix, such as calcium and phosphate. In the present work, the mobility of tungsten compounds was investigated in a model soil with a range of aqueous leach solutions using column experiments. The relative column leachate concentrations measured followed trends from previously reported tungstate and polytungstate partition coefficients determined in the model soil under identical aqueous matrix conditions. Neutral to alkaline conditions produced maximum effluent tungsten concentrations >40 mgL(-1), whereas acid leach eluents produced concentrations in the <1-3 mgL(-1) range. The change in leached tungsten speciation over time was also measured as monomeric and polymeric tungsten species have different sorptive behaviors.

Comparison of standard and reaction cell inductively coupled plasma mass spectrometry in the determination of chromium and selenium species by HPLC-ICP-MS.

Elemental speciation is becoming a common analytical procedure for geochemical investigations. The various redox species of environmentally relevant metals can have vastly different biogeochemical properties, including sorption, solubility, bioavailability, and toxicity. The use of high performance liquid chromatography (HPLC) coupled to elemental specific detectors, such as inductively coupled plasma mass spectrometry (ICP-MS), has become one of the most important speciation methods employed. This is due to the separation versatility of HPLC and the sensitive and selective detection capabilities of ICP-MS. The current study compares standard mode ICP-MS to recently developed reaction cell (RC) ICP-MS, which has the ability to remove or reduce many common polyatomic interferences that can limit the ability of ICP-MS to quantitate certain analytes in complex matrices. Determination of chromium and selenium redox species is achieved using ion-exchange chromatography with elemental detection by standard and RC-ICP-MS, using various chromium and selenium isotopes. In this study, method performance and detection limits for the various permutations of the method (isotope monitored or ICP-MS detection mode) were found to be comparable and generally less than 1 microg L(-1). The method was tested on synthetic laboratory samples, surface water, groundwater, and municipal tap water matrices.

Geochemical parameters influencing tungsten mobility in soils.

The biogeochemistry of tungsten and its effects on mobility have recently gained attention due to the existence of human cancer clusters, such as in Fallon, NV. Tungsten exists in many environmental matrices as the soluble and mobile tungstate anion. However, tungsten can polymerize with itself and other anions, creating poly- and heteropoly-tungstates with variable geochemical and toxicological properties. In the present work, geochemical parameters are determined for tungstate species in a model soil that describe the potential for tungsten mobility. Soluble tungsten leached from a metallic tungsten-spiked soil after six to twelve months aging reached an equilibrium concentration >150 mg/L within 4 h of extraction with deionized water. Partition coefficients determined for various tungstate and polytungstate compounds in the model soil suggest a dynamic system in which speciation changes over time affect tungsten geochemical behavior. Partition coefficients for tungstate and some poly-species have been observed to increase by a factor of 3 to 6 over a four month period, indicating decreased mobility with soil aging.

Sieve element specific labeling with an anti-idiotypic monoclonal antibody mimic of the phytotoxin dothistromin.

A monoclonal antibody, 12C9, an anti-idiotypic mimic of dothistromin, a toxin produced by Dothistroma pini, was found to label the cell wall of sieve elements in a number of different plant tissues and species. The antibody labeled apple leaf tissue, tobacco leaf mid vein, leaf and meristem, and Coprosma robusta leaf mid vein. Labeling was restricted to cell walls of sieve elements and did not label the companion cells or the lumen of the cells. The antibody labeled over a wide range of dilutions. This antibody could be used to differentiate sieve elements from other types of phloem. It could also be used to co-localize sieve elements and microorganisms such as phytoplasmas stained with DAPI.

Effects of time and temperature during attachment of sections to microscope slides on immunohistochemical detection of antigens.

To study the effects of time and temperature on attachment of tissue sections to microscope slides, we examined the intensity of immunohistochemical staining of selected antigens in nine different neoplastic and normal tissues after attaching sections at different times and temperatures. Typically, both the temperature and time are minimized when tissue sections attached to slides; however, suboptimal times and temperatures during attachment may result in either loss of tissue due to poor attachment or the necessity for inconvenient staining regimens. Using standard immunohistochemical techniques, 5 microm tissue sections were attached at 58 degrees C for 1, 4 and 24 hr. In a separate study, 5 microm tissue sections were attached for 16 hr at 58, 68 and 80 degrees C. The intensity of staining decreased slightly when the tissue sections were heated at 80 degrees C for 16 hr, but there was little or no decrease when tissues were heated at 68 degrees C or lower for 16 hr, or at 58 degrees C for up to 24 hr.

A data mining system for infection control surveillance.

Nosocomial infections and antimicrobial resistance are problems of enormous magnitude that impact the morbidity and mortality of hospitalized patients as well as their cost of care. The Data Mining Surveillance System (DMSS) uses novel data mining techniques to discover unsuspected, useful patterns of nosocomial infections and antimicrobial resistance from the analysis of hospital laboratory data. This report details a mature version of DMSS as well as an experiment in which DMSS was used to analyze all inpatient culture data, collected over 15 months at the University of Alabama at Birmingham Hospital.

Application of data mining to intensive care unit microbiologic data.

We describe refinements to and new experimental applications of the Data Mining Surveillance System (DMSS), which uses a large electronic health-care database for monitoring emerging infections and antimicrobial resistance. For example, information from DMSS can indicate potentially important shifts in infection and antimicrobial resistance patterns in the intensive care units of a single health-care facility.

Association rules and data mining in hospital infection control and public health surveillance.

OBJECTIVES: The authors consider the problem of identifying new, unexpected, and interesting patterns in hospital infection control and public health surveillance data and present a new data analysis process and system based on association rules to address this problem. DESIGN: The authors first illustrate the need for automated pattern discovery and data mining in hospital infection control and public health surveillance. Next, they define association rules, explain how those rules can be used in surveillance, and present a novel process and system--the Data Mining Surveillance System (DMSS)--that utilize association rules to identify new and interesting patterns in surveillance data. RESULTS: Experimental results were obtained using DMSS to analyze Pseudomonas aeruginosa infection control data collected over one year (1996) at University of Alabama at Birmingham Hospital. Experiments using one-, three-, and six-month time partitions yielded 34, 57, and 28 statistically significant events, respectively. Although not all statistically significant events are clinically significant, a subset of events generated in each analysis indicated potentially significant shifts in the occurrence of infection or antimicrobial resistance patterns of P. aeruginosa. CONCLUSION: The new process and system are efficient and effective in identifying new, unexpected, and interesting patterns in surveillance data. The clinical relevance and utility of this process await the results of prospective studies currently in progress.

Syntheses of haptens containing dioxaphosphorinan methoxyacetic acid linker arms for the production of antibodies to organophosphate pesticides.

Four generic heterobifunctional reagents, namely 2-(2-chloro-5-methyl-1,3,2-dioxaphosphorinan-5-yl)methoxyacetic acid methyl ester, p-sulfide, 2-(2-chloro-5-methyl-1,3,2-dioxaphosphorinan-5-yl)-methoxyacetic acid methyl ester, p-oxide, 2-(2-mercapto-5-methyl-1,3,2-dioxaphosphorinan-5-yl)-methoxyacetic acid bispotassium salt, p-sulfide-, and (2-methoxy-5-methyl-1,3,2-dioxaphosphorinan-5-yl)methoxyacetic acid, methyl ester, have been synthesized and used to prepare organophosphate, thiophosphate, and dithiophosphate haptens containing a functional carboxyl group which can be used to conjugate the haptens to proteins. These hapten-protein conjugates have been used as antigens for preparing polyclonal sera against all classes of organophosphate pesticides. The eight examples used protein-hapten conjugates of chlorpyrifos, parathion, diazinon, paraoxon, azinphos, dimethoate, demeton, and dichlorvos. These were all immunogenic and resulted in sera containing antibodies that recognized the corresponding parent pesticide with high specificity.

A system for tissue-specific copper-controllable gene expression in transgenic plants: nodule-specific antisense of aspartate aminotransferase-P2.

A vector system, based on copper controllable gene expression, has been developed to give control over place as well as time of expression of an introduced gene. This system consists of two elements: (1) the yeast ace1 gene encoding a metallo-regulatory transcription factor, ACE1, under control of either an organ-specific or a constitutive promoter; and (2) a gene of interest under control of a chimaeric promoter consisting of the 46 bp TATA fragment of the CaMV 35S RNA promoter linked to four repeats of the ACE1 binding site. The functioning of the system in an organ-specific manner was tested in nodulated Lotus corniculatus plants which consisted of non-transformed shoots plus transformed hairy root tissue 'wild-type tops/transgenic roots'. After addition of copper ions to the plant nutrient solution, beta-glucuronidase (GUS) expression was visualized either specifically in nodules or in both roots and nodules when the ace1 gene was placed under control of the nod45 promoter or the CaMV 35S RNA promoter, respectively. The nodule-specific system was used to express antisense constructs of aspartate aminotransferase-P2 in transgenic Lotus corniculatus plants. When expression was induced by the addition of copper ions to the plant nutrient solution aspartate aminotransferase-P2 activity declined dramatically, and a decrease of up to 90% was observed in nodule asparagine concentration.

An Internet health care information resources server as a component of a statewide medical information network.

A health care information server utilizing Internet resource discovery technology is presented as a component of a statewide medical information network. The development of an information server, including the development process and its design and operation, is presented. Menu design and implementation, which involved providing access to information resources in support of the tasks that make up the health care delivery process, are described. The potential impact of this technology on the health care delivery process is explored, and ways in which access to information can be facilitated and matched with the information needs of the various health care delivery tasks are identified. Issues associated with the use of public domain information resources are discussed, including control over Internet resources, access to information resources, network operational delays, client connection and software availability, information quality, and menu navigation. This project has demonstrated that Internet information resources exist that match the information needs of the tasks that make up the health care delivery process. Positive response has been received from physicians after initial utilization of the server in a stand-alone context. In the future, more applications will integrate the vast information resources on the Internet with traditional computing systems.

Cloning and characterization of a cDNA encoding aspartate aminotransferase-P1 from Lupinus angustifolius root tips.

A root tip cDNA library, constructed in the lambda Zap II expression vector, was immunoscreened with a monoclonal antibody raised against aspartate aminotransferase-P1 from Lupinus angustifolius L. var Uniharvest. One 1452-base pair clone was isolated. The encoded protein sequence had high homology to both plant and animal aspartate aminotransferase sequences. The clone was converted to the phagemid form and expressed in Escherichia coli. The expressed protein was enzymically active and could be immunocomplexed with aspartate aminotransferase-P1-specific antibodies. The complete aspartate aminotransferase-P1 cDNA was cloned into the yeast expression vector pEMBL-yex4 and transformed into Saccharomyces cerevisiae strain BRSCS6, which possesses a mutated aspartate aminotransferase gene (the asp5 mutation). Complementation of the mutation was achieved using this construct.

Providing access to healthcare information resources using Internet Gopher technology as a part of a state-wide medical information network.

An Internet healthcare information resources Gopher server is described as a part of a state-wide medical information network. The development of the server and its design and operation are presented. The potential impact of this technology on the healthcare delivery process and issues associated with the use of public domain information resources are discussed.

Production and Characterization of Monoclonal Antibodies against Aspartate Aminotransferase-P1 from Lupin Root Nodules.

Six hybridoma clones were obtained that secreted monoclonal antibodies against the aspartate aminotransferase-P1 (AAT-P1) isoenzyme from root nodules of Lupinus angustifolius [L.] cv Uniharvest. This enzyme is found constitutively in the plant cytosol fraction. The monoclonal antibodies produced were all of the immunoglobulin G1 class, recognized two distinct epitopes on the protein, and represented the major paratopes found in the immunoglobulin fraction of sera taken from mice and rabbits immunized with the pure AAT-P1 protein. One of these epitopes was unique to lupin nodule AAT-P1. The other epitope was shown to be present on enzyme from lupin bean, white clover and tobacco leaves, lupin roots and nodules, and potato tubers. Both epitopes were recognized by the appropriate monoclonal antibodies in both their native and denatured forms. None of the monoclonal antibodies produced reacted with Rhizobium lupini NZP2257, Escherichia coli extracts, or with the inducible aspartate aminotransferase-P2 (AAT-P2) isoform also found in root nodules. A sandwich enzyme-linked immunosorbent assay utilizing two monoclonal antibodies recognizing the two distinct epitopes was developed and was capable of quantitating AAT-P1 in plant extracts. The limit of detection of AAT-P1 was less than 15 pg/mL and AAT-P1 protein could be quantified in the range 80 to 1000 pg/mL. Using this assay, AAT-P1 protein was shown to remain relatively constant during nodule development. Use of an AAT-P2-specific monoclonal antibody that inhibits the enzyme activity of this isoform enabled the direct determination of AAT-P1 enzyme activity in nodule extracts. Using these assays, specific activities of the individual isoforms were calculated; that of the AAT-P1 isoform was shown to be 7.5-fold higher than that of the AAT-P2 isoform.

Production of monoclonal antibodies recognising different epitopes present on insulin-like growth factor 1.

Monoclonal antibodies (MAbs) were generated against recombinant human insulin-like growth factor 1 (IGF-1) by fusion of NS-1 myeloma cells with spleen cells from BALB/c X DBA mice immunised with recombinant IGF-1 and synthetic peptide sequences derived from the published amino acid sequence of IGF-1. MAbs were produced that recognised four distinct epitopes including two defined segments of the C and D domains. All MAbs were IgM mouse immunoglobulins. These results indicate the feasibility of producing MAbs to highly conserved proteins.