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A procedure for the rapid detection of depleted uranium in metal shrapnel fragments.

Depleted uranium is now widely used in the armor of military vehicles as well as in kinetic-energy penetrators designed to defeat enemy armor. As a result, the potential that personnel will be wounded by depleted uranium fragments has increased. Because toxicities associated with depleted uranium fragments may ultimately require different treatment protocols than those used for traditional metal fragment injuries, a method to rapidly detect the presence of depleted uranium in surgically excised shrapnel fragments is required. By treating the shrapnel fragment with an extracting agent, such as nitric acid, for 5 minutes in an ultrasonic cleaner, sufficient metal is solubilized to allow for colorimetric detection using a pyridylazo dye. Although several metals are capable of being detected under these conditions, the reaction can be made specific for depleted uranium through the use of masking agents such as sodium citrate and ethylenediaminetetraacetic acid. This procedure allows for the rapid (< 15 minutes) extraction and detection of depleted uranium in metal shrapnel fragments.

Azo Compounds↗

Rapid determination of 226Ra and uranium isotopes in solid samples by fusion with lithium metaborate and alpha spectrometry.

A simple and rapid method has been developed to determine 226Ra in rocks, soils, and sediments. Samples are decomposed by fusion with lithium metaborate and the melt is dissolved in a solution containing sulfates and citric acid. During the dissolution, a fine suspension of mixed barium and radium sulfates is formed. The microcrystals are collected on a membrane filter (pore size 0.1 microm) and analysed in an alpha spectrometer. Application of a 133Ba tracer enables us to assess the loss of the analyte, which only rarely exceeds 10%. All analytical operations, beginning from sample decomposition to source preparation for alpha spectrometry, can be accomplished within 1 or 2 h. With uranium determination, the filtrate is spiked with a 232U tracer and passed through a column loaded with a Dowex AG (1 x 4) anion-exchange resin in the sulfate form. Interfering elements are eluted with dilute sulfuric acid followed by concentrated hydrochloric acid. Uranium is eluted with water, electrodeposited on silver discs, and analysed in the alpha spectrometer. The method was tested on reference soil and sediment materials and was found to be accurate within the estimated uncertainties.

Borates↗

A new procedure for the spectrophotometric determination of uranium(VI) in the presence of a large excess of thorium(IV).

Spectrophotometric determination of microgram amounts of U(VI) with 2-(5-Bromo-2-Pyridylazo)-5-diethylaminophenol (Br-PADAP), originally developed by Johnson and Florence has been modified to enable the determination of U(VI) in the presence of a large excess of Th(IV). The effects of thorium, tri-n-butyl phosphate (TBP) and ethanol on the estimation of uranium have been studied in detail and are presented in this paper. This modified method can be applied for the analysis of U(VI) both in aqueous and organic samples containing a large excess of Th(IV) (Th:U = 10000:1).

Azo Compounds↗

Dioxouranium(VI)--carboxylate complexes. Interaction with dicarboxylic acids in aqueous solution: speciation and structure.

In this paper we report the results of an investigation performed by potentiometric (H+-glass electrode) and visible spectrophotometric measurements on the interaction of UO2(2+) ion towards some carboxylic ligands (acetate, malonate, succinate, azelate). The measurements were carried out at T= 25 degrees C in different ionic media (KNO3 and NaCl) at different ionic strengths (0.1 < or = I/mol L(-1) < or = 1.0, NaCl; I/mol L(-1) = 0.1, KNO3). The dependence on ionic strength of formation constants was taken into account by using both a simple Debye-Hückel type equation and the SIT (Specific ion Interaction Theory) approach. Different speciation models (depending on concentration of reagents, ionic strength, pH-range) both for different carboxylates and different ionic media have been obtained. Linear combinations between formation constants, stoichiometric coefficients and length of alkyl chain of dicarboxylates have been observed and predicted formation constants at I= 0 mol L(-1) are reported for the interaction of UO2(2+) with HOOC-(CH2)n-COOH with 1 < or = n < or = 7. Finally, a visible absorption spectrum for each complex reaching a significant percentage of formation in solution (KNO3 medium) has been calculated to characterise the compounds found by pH-metric refinement.

Dicarboxylic Acids↗

The use of radioimmune assay in investigating reagents to be used in the immunocytochemical localization of hepatitis B surface antigen in immune complexes in the kidney of patients with membranous nephropathy and Australia antigenaemia.

Radioimmune assay (RIA) was used to investigate the effect of fixatives on antigenicity of the hepatitis B surface antigen (HBsAg) and the effect of pronase on the elution of antibody (Ab) from the HBsAG-Ab complex. The effect of pronase on Ab elution was also tested on sections of kidney from a patient with the immune complex disease systemic lupus erythematosus (SLE). Immunoglobulin G (IgG) was located in pronase treated and untreated sections using the indirect immunoperoxidase technique. Glutaraldehyde was shown to be the fixative of choice for studies involving HbsAG. All fixatives were shown to have less effect on antigenicity at 4 degrees C than at room temperature. Osmium tetroxide reduced antigenicity to one-third, even at 4 degrees C. RIA and SLE kidney section studies showed that Ab was eluted from immune complexes by pronase. Pre-fixation of the antigen (Ag) by glutaraldehyde appears to have no effect on the final elution, although fixation after pronase treatment seemed to enhance the elution effects. The availability of an RIA kit with HBsAg- and AB-coated beads was of great assistance in evaluating reagents to be used in immunoperoxidase studies of HBsAg in immune complexes of patients with membranous nephropathy and Australia antigenaemia.

Antigen-Antibody Complex↗

Negative staining of rat tail tendon collagen fibrils with uranyl formate.

Negative staining of rat tail tendon collagen fibrils with uranyl formate appears to reveal more detail in the axial banding pattern than any other positive or negative staining method hitherto employed. In addition, uranyl formate and other uranyl solutions appear to reveal fine, closely spaced, longitudinal filaments which may represent the individual tropocollagen molecules.

Acetates↗

Chelation in metal intoxication. XIV. Comparative effect of thiol and amino chelators on lead-poisoned rats with normal or damaged kidneys.

D-Penicillamine (DPA), diethyldithiocarbamate (DDC), L-cysteine, ethylenediaminetetraacetic acid (EDTA), cyclohexylenediaminetetraacetic acid (CDTA), and diethylene triamine pentaacetic acid (DTPA) were compared for their efficacy to enhance urinary excretion of Pb, to reduce Pb concentration of body organs, and to restore the enhanced urinary excretion of delta-aminolevulinic acid (delta-ALA), the inhibited activities of blood delta-ALA dehydratase, and renal enzymes in Pb-administered rats (10 mg/kg, po, 4 weeks) with normal or experimentally damaged kidneys. The acute renal damage was induced by uranyl acetate (3 mg/kg, sc, once) prior to treatment with the chelators (0.3 mmol/kg, ip, twice) and evaluated by enhanced urinary excretion of diagnostic enzymes and inhibition in their renal activities. Among thiol chelators, DPA was the most effective followed by DDC in enhancing the urinary excretion of Pb, reducing the concentration of Pb in blood, kidneys and liver, and in restoring Pb-induced biological alterations in urine, blood, and kidneys. Among amino carboxylic acids, DTPA was the most effective and EDTA and CDTA were about equally potent in countering Pb toxicity. Protection was more marked in animals with normal kidneys than in those with acutely damaged kidneys.

Alkaline Phosphatase↗

Analysis of chromosomal aberration frequencies in the peripheral blood lymphocytes of smokers exposed to uranyl compounds.

One hundred fifteen smokers working in a nuclear fuel manufacturing facility were analysed for various types of chromosomal aberrations. They experienced exposure for a period of 1-25 years. Their age ranges from 23 to 52 years. A total of 94 smokers and 118 non-smokers who were not exposed to uranyl compounds or to any other known mutagens and belong to the same age group formed the control subjects. The results showed that there is a significant increase in the frequency of chromosomal aberrations in the exposed smokers when compared to the control smokers. In the control group, the smokers showed a high frequency of chromosomal aberrations when compared to non-smokers suggesting clastogenic effect of smoking. Chromosomal aberrations observed in the exposed smokers could be due to the cumulative effect of both smoking and exposure to uranyl compounds.

Adult↗

The covalent and three-dimensional structure of concanavalin A.

The tentative amino-acid sequence and three-dimensional structure of the lectin concanavalin A have been determined. The amino-acid sequence, which was determined chemically, contains 238 residues. The sequences of three short stretches were assigned on the basis of x-ray crystallographic data. Interpretation of an electron density map at 2-A resolution indicates that the predominant structural element is extended polypeptide chain arranged in two anti-parallel pleated sheets or beta-structures. Residues not included in the beta-structures are arranged in regions of random coil. One of the pleated sheets contributes extensively to the interactions among the monomers to form both dimers and tetramers. The positions at which Mn(2+), Ca(2+), and saccharide are bound to the protein, and the point of cleavage for the formation of the naturally occurring fragments A(1) and A(2), have been tentatively assigned. Both metal-binding sites are at least 20-A removed from the position at which saccharides are bound. The saccharide-binding site is a deep pocket of approximately 6A x 7.5A x 18A, the inner portion of which is occupied by hydrophobic residues.

Amino Acid Sequence↗

Electron microscopy of negatively stained and unstained fibrinogen.

Electron microscope images of negatively stained fibrinogen are predominantly asymmetric rods 450 A in length and about 60 A in width. The molecules appear to have considerable flexibility, and mass distribution along the major axis is not uniquely distinguished despite apparent beading in some particles. Scanning transmission electron microscopy of unstained fibrinogen again demonstrates that a majority of molecules are rodlike. The results differ from those obtained by negative staining in that a substantial fraction of images are trinodular with striking resemblance to those obtained by C. E. Hall and H. S. Slayter [J. Biophys. Biochem. Cytol. (1959) 5, 11--16] using the mica replica technique. The above results were obtained on glow-discharged carbon substrate films by a simple low-concentration, long-attachment-time modification of standard deposition methods that is diffusion controlled and depends on concentration and time but is independent of pH, buffer, and other staining conditions. Evidence is presented that standard attachment procedures result in artifactual images. Any models of fibrinogen in solution consequently must encompass properties that permit its visualization as an asymmetetric rod by electron microscopy as first suggested by Hall and Slayter 20 years ago.

Animals↗

Staining of intracellular deposits of uranium in cultured murine macrophages.

In our studies of the health effects of internalized depleted uranium, we developed a simple and rapid light microscopic method to stain specifically intracellular uranium deposits. Using J774 cells, a mouse macrophage line, treated with uranyl nitrate and the pyridylazo dye 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol, uranium uptake by the cells was followed. Specificity of the stain for uranium was accomplished by using masking agents to prevent the interaction of the stain with other metals. Prestaining wash consisting of a mixture of sodium citrate and ethylenediaminetetraacetic acid eliminated staining of metals other than uranium. The staining solution consisted of the pyridylazo dye in borate buffer along with a quaternary ammonium salt, ethylhexadecyldimethylammonium bromide, and the aforementioned sodium citrate/ethylenediaminetetraacetic acid mixture. The buffer was essential for maintaining the pH within the optimum range of 8 to 12, and the quaternary ammonium salt prevented precipitation of the dye. Staining was conducted at room temperature and was complete in 30 min. Staining intensity correlated with both uranyl nitrate concentration and incubation time. Our method provides a simple procedure for detecting intracellular uranium deposits in macrophages.

Animals↗

XAFS determination of the bacterial cell wall functional groups responsible for complexation of Cd and U as a function of pH.

Bacteria, which are ubiquitous in near-surface geologic systems, can affect the distribution and fate of metals in these systems through adsorption reactions between the metals and bacterial cell walls. Recently, Fein et al. (1997) developed a chemical equilibrium approach to quantify metal adsorption onto cell walls, treating the sorption as a surface complexation phenomenon. However, such models are based on circumstantial bulk adsorption evidence only, and the nature and mechanism of metal binding to cell walls for each metal system have not been determined spectroscopically. The results of XAFS measurements at the Cd K-edge and U L3-edge on Bacillus subtilis exposed to these elements show that, at low pH, U binds to phosphoryl groups while Cd binds to carboxyl functional groups.

Adsorption↗

A new technique for perfusion fixation and contrast enhancement of foetal lamb myocardium for electron microscopy.

A technique of perfusion fixation following elective diastolic cardiac arrest was developed to obtain excellent preservation of foetal lamb hearts for morphological studies with electron microscopy. A technique was developed to markedly enhance the contrast of embedded material by treating thin sections with tannic acid prior to lead staining. This technique has been particularly useful for quantitative morphometric analysis.

Animals↗

A routine method for contrasting elastin at the ultrastructural level.

A reliable, simple, and inexpensive method for ultrastructural investigation of elastin is described. This method uses uranyl acetate dissolved in absolute methanol, followed by an optional lead citrate counterstain. The procedure was tested on a number of animal and human tissues that had been fixed and processed differently.

Animals↗