Rapid quantitative measurements of proteomes by Fourier transform ion cyclotron resonance mass spectrometry.

The patterns of gene expression, post-translational modifications, protein/biomolecular interactions, and how these may be affected by changes in the environment, cannot be accurately predicted from DNA sequences. Approaches for proteome characterization are generally based upon mass spectrometric analysis of in-gel digested two dimensional polyacrylamide gel electrophoresis (2-D PAGE) separated proteins, allowing relatively rapid protein identification compared to conventional approaches. This technique, however, is constrained by the speed of the 2-D PAGE separations, the sensitivity limits intrinsic to staining necessary for protein visualization, the speed and sensitivity of subsequent mass spectrometric analyses for identification, and the limited ability for accurate quantitative measurements based on differences in spot intensity. We are presently developing alternative approaches for proteomics based upon the combination of fast capillary electrophoresis, or other suitable chromatographic separations, and the high mass accuracy and sensitivity obtainable with unique Fourier transform ion cyclotron resonance (FTICR) mass spectrometers available at our laboratory. Several approaches are presently being pursued; one based upon the analysis of intact proteins and the second upon approaches for global protein digestion and accurate peptide mass analysis. Quantitation of protein/peptide levels are based on using two or more stable-isotope labeled versions of proteomes which are combined to obtain precise quantitation of relative protein abundances. We describe the status of our efforts towards the development of a high-throughput proteomics capability and present initial results for application to several microorganisms and discuss our efforts for extending the developed capability to mammalian proteomes.     

Self contained enrichment cell for tritium

Tritiated water is probably the most mobile manmade nuclide found in the environment. The low-energy beta decay produces few health concerns except at extremely high concentrations of tritium. However, because of the ubiquitous nature of tritiated water, when identified in the bionetwork, it can be viewed as a precursor of nuclides with greater health risks. For this reason alone it becomes desirable to determine tritium concentrations many orders of magnitude below that considered a health concern. A devise is described that eliminates the need for large water-filled chest freezers used as coolers. The enrichment cells can be employed individually or in series.     

Radiological characterization of neutron-activated metal wastes from nuclear power stations

Three different methods for determining the long-lived radionuclide contents of highly neutron activated metal wastes are being compared and assessed using samples of pressurized-water reactor (PWR) and boiling-water reactor (BWR) spent fuel disassembly hardware and control rods. These methods include: 1) empirical analyses involving sampling and laboratory measurements; 2) direct assay techniques; 3) calculation methods. These techniques are discussed and the results of the comparisons made to date are presented. The agreement between empirical versus calculational methods for the disassembly hardware was generally within 10% to 50%, and rarely exceeded a factor of 2.5, especially near the fueled region of the reactor core. However, large discrepancies between the measured versus calculated concentrations were observed for⁵⁹Ni and⁶³Ni at the end fittings of the fuel assembly hardware, where the calculations underestimated the concentrations by factors varying from about 8 to 28. The calculation errors are believed to be due to inadequate cross section data for the nickel isotopes.Pacific Northwest Laboratory is operated for the U. S. Department of Energy by Battelle Memorial Institute under Contract DE-AC06-76RLO 1830.     

Stochastic radiative transfer in multilayer broken clouds. Part I: Markovian approach

A two-part paper describes the statistical treatment of solar radiative transfer in multilayer broken clouds. The proposed approach is a logical development of the statistical ones originally suggested for a single-layer broken clouds. This first part introduces a new statistically inhomogeneous Markovian model that allows one to properly account for different combinations of the random and maximum overlap of broken clouds at distinct vertical layers. The statistically inhomogeneous Markovian model and the stochastic radiative transfer equation have been used to derive equations for the mean radiance of solar radiation. It was demonstrated that in extreme cases the obtained equations agree with corresponding equations previously derived for (i) the statistically homogeneous broken clouds and (ii) the vertically inhomogeneous overcast clouds.     

Creation of variable concentrations of defects on TiO2(110) using low-density electron beams

Low density (˜μA/cm²) 0.48 and 1.0 keV electron beams have been used to create surface defects on a TiO₂(110) surface. These electron-beam induced defects were examined primarily by X-ray photoelectron spectroscopy (XPS) with supporting ultraviolet photoemission spectroscopy (UPS). Glancing and normal emission XPS spectra of nearly defect-free surfaces revealed that Ti atoms on the surface were similar to the bulk Ti, while some surface oxygen atoms were different from the bulk oxygen. XPS of Ti 2p_3/2 was used to quantify the defect concentration and to examine the defect electronic structure. Based on our calculation of defect concentrations and the comparison of our results with results and models from the literature, we conclude that oxygen vacancies induced by electron beams in the current study are mostly from the bridging oxygen sites, in agreement with the previous work. A range of defect concentrations with similar electronic structure, mainly composed of Ti³⁺, have been induced by low-density electron beams. Beam energy and exposure were the experimental variables. The rates of defect formation at low beam exposure were beam-energy dependent, with a faster growth rate at 0.48 keV than at 1.0 keV. These defects were similar to those by thermal annealing in vacuum, but a higher concentration of defects could be obtained with longer beam exposure. However, the e-beam induced defects were different from those produced by Ar⁺ ion bombardment since both this and previous studies have found defects produced by Ar⁺ ion bombardment to be complex, with a variety of different local environments where oxygen and titanium surface atoms coexist.     

The measurement of222Rn in drinking water by low-level liquid scintillation counting

Radon-222 (Rn) has universally been found in well water. Non-stagnant ground water is collected at the well head while the well is pumping. The water is adjusted to a slow, non-aerated, steady flow through a clear tube, and a 500 ml glass bottle is filled. The sample is tightly capped after a high meniscus has developed. In the laboratory, standard 22 ml glass vials are filled with 10 ml of a toluene based mineral oil LS cocktail. Then, two 5 ml sample aliquots are pipetted into the vial. Vials are capped tightly, shaken vigorously, and placed in the liquid scintillation (LS) counter. Secular equilibrium is established in approximately 4 hours, after which samples are counted for 100 minutes each. The counting efficiency for Rn and progeny ranges between 315 to 345 percent depending on the chosen spectral window. The average background is about 6 cpm. A total of 28 wells were tested for Rn in the Carefree-Cave Creek, Arizona, USA area. The area's geometric average Rn concentration was found to be 46.5 Bq·l^–1. The associated estimated lung dose is 0.51 mSv·y^–1.Deceased 1 June 1991.     

Experimental limitations in impedance spectroscopy: Part VI. Four-point measurements of solid materials systems

Four-point impedance spectroscopy of solid materials systems is severely hampered by unavoidable voltage-divider effects associated with the reference electrodes. As demonstrated by test circuit studies and experiments with Pt/YSZ/Pt cells (with embedded silver reference electrodes), high impedance reference electrodes can produce distorted and erroneous impedance data. The relationships between these data and the sample properties (conductivity, dielectric constant) and instrument limitations (input impedance/capacitance) are derived. Successful four-point impedance measurements on conductive systems require large effective dielectric constants, which may be unattainable in bulk solids, but are often associated with internal interfaces.