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Centrosomes nucleate and organize interphase MTs and areinstrumental in the assembly of the mitotic bipolar spindle. Here wereport that two members of the multifunctional protein 4.1 family havedistinct distributions at centrosomes. Protein 4.1R localizes to maturecentrioles whereas 4.1G is a component of the pericentriolar matrixsurrounding centrioles. To selectively probe 4.1R function, we used RNAinterference-mediated depletion of 4.1R without decreasing 4.1Gexpression. 4.1R downregulation reduces MT anchoring and organization atinterphase and impairs centrosome separation during prometaphase.Metaphase chromosomes fail to properly condense/align and spindleorganization is aberrant. Notably 4.1R depletion causes mislocalizationof its binding partner NuMA (Nuclear Mitotic Apparatus Protein),essential for spindle pole focusing, and disrupts ninein. Duringanaphase/telophase, 4.1R-depleted cells have lagging chromosomes andaberrant MT bridges. Our data provide functional evidence that 4.1R makescrucial contributions to centrosome integrity and to mitotic spindlestructure enabling mitosis and anaphase to proceed with the coordinatedprecision required to avoid pathological events.

The central metabolic fluxes of Shewanella oneidensis MR-1were examined under carbon-limited (aerobic) and oxygen-limited(micro-aerobic) chemostat conditions using 13C labeled lactate as thesole carbon source. The carbon labeling patterns of key amino acids inbiomass were probed using both GC-MS and 13C-NMR. Based on the genomeannotation, a metabolic pathway model was constructed to quantify thecentral metabolic flux distributions. The model showed that thetricarboxylic acid (TCA) cycle is the major carbon metabolism route underboth conditions. The Entner-Doudoroff and pentose phosphate pathways weremainly utilized for biomass synthesis (flux below 5 percent of thelactate uptake rate). The anapleurotic reactions (pyruvate to malate andoxaloacetate to phosphoenolpyruvate) and the glyoxylate shunt wereactive. Under carbon-limited conditions, a substantial amount of carbonwas oxidized via the highly reversible serine metabolic pathway. Fluxesthrough the TCA cycle were less whereas acetate production was more underoxygen limitation than under carbon limitation. Although fluxdistributions under aerobic, micro-aerobic, and shake-flask cultureconditions were dramatically different, the relative flux ratios of thecentral metabolic reactions did not vary significantly. Hence, S.oneidensis metabolism appears to be quite robust to environmentalchanges. Our study also demonstrates the merit of coupling GC-MS with 13CNMR for metabolic flux analysis to reduce the use of 13C labeledsubstrates and to obtain more accurate flux values.

Model evolution and improvement is complicated by the lack of high quality observational data. To address a major limitation of these measurements the Atmospheric Radiation Measurement (ARM) program was formed. For the second quarter ARM metric we will make use of new water vapor data that has become available, and called the 'Merged-sounding' value added product (referred to as OBS, within the text) at three sites: the North Slope of Alaska (NSA), Darwin Australia (DAR) and the Southern Great Plains (SGP) and compare these observations to model forecast data. Two time periods will be analyzed March 2000 for the SGP and October 2004 for both DAR and NSA. The merged-sounding data have been interpolated to 37 pressure levels (e.g., from 1000hPa to 100hPa at 25hPa increments) and time averaged to 3 hourly data for direct comparison to our model output.

The ability of wetted cathodes of limited area to support localized corrosion sites on passive materials exposed to atmospheric conditions was studied computationally. The analysis pertains to conditions where metal surfaces are covered by thin layers of moisture in contrast to conditions of full immersion. The moisture may be a continuous layer or in patches with and without particulate on the surface. These conditions are of interest for the surfaces of the waste packages at the proposed Yucca Mountain Repository where waste packages are supported in air. The cathode capacity was characterized by the total net cathodic current, I{sub net}, which the surface surrounding a localized corrosion site (i.e., a pit or crevice) could supply. The cathode capacity increases with increasing cathode area, but it saturates at finite cathode sizes due to the resistance of the thin electrolyte layer. The magnitude of the capacity depends on the water layer thickness, the solution conductivity, and the electrochemical reaction kinetics. The presence of particulates is treated by considering both volume and surface coverage effects. The limited electrolyte volume under thin film conditions can lead to rapid pH changes which decrease the cathode capacity due to the slower electrochemical kinetics at elevated pH. …

Sorption of {sup 237}Np(V) and {sup 233}U(VI) was measured on clays separated from Yucca Mountain alluvium as a function of solution pH and aqueous actinide concentrations. The results indicate that sorption of U and Np on the separated clay fraction depends strongly on solution pH. Np sorption on clays increases slowly with increasing pH from 3 to 7. Above pH 7, Np sorption on clays increases rapidly up to a pH of about 10. On the other hand, U sorption on clays reaches it maximum at a pH of about 6, with sorption decreasing as pH increases from 6 to 8 and then increasing again as pH increases further from 8 to about 10. The results suggest that a Freundlich isotherm can be used to describe U and Np sorption on clays at pH above 5.5. The results of this study indicate that clay minerals play a very important role in the sorption of U and Np on Yucca Mountain alluvium. Indeed, the clay content of the alluvium is probably considerably more important than water chemistry in predicting the ability of the alluvium to attenuate the transport of these radionuclides.

Uranium-series ages, oxygen-isotopic compositions, and uranium contents were determined in outer growth layers of opal and calcite from 0.5- to 3-centimeter-thick mineral coatings hosted by lithophysal cavities in the unsaturated zone at Yucca Mountain, Nevada, the proposed site of a permanent repository for high-level radioactive waste. Micrometer-scale growth layering in the minerals was imaged using a cathodoluminescence detector on a scanning electron microscope. Determinations of the chemistry, ages, and delta oxygen-18 values of the growth layers were conducted by electron microprobe analysis and secondary ion mass spectrometry techniques at spatial resolutions of 1 to about 20 micrometers ({micro}m) and 25 to 40 micrometers, respectively. Growth rates for the last 300 thousand years (k.y.) calculated from about 300 new high-resolution uranium-series ages range from approximately 0.5 to 1.5 {micro}m/k.y. for 1- to 3-centimeter-thick coatings, whereas coatings less than about I-centimeter-thick have growth rates less than 0.5 {micro}m/k.y. At the depth of the proposed repository, correlations of uranium concentration and delta oxygen-18 values with regional climate records indicate that unsaturated zone percolation and seepage water chemistries have responded to changes in climate during the last several hundred thousand years.

{sup 99}Tc and {sup 129}I are important contributors to risk assessment due to their long half-lives and high mobility as aqueous anionic species. We analyzed {sup 99}Tc and {sup 129}I in groundwater samples in and near 11 underground nuclear tests and in melt glass and rock samples retrieved from the Chancellor test cavity, Nevada Test Site. The {sup 129}I/{sup 127}I ratio ranges from 10{sup -3} to 10{sup -6} in cavity water and 10{sup -4} to 10{sup -9} in satellite wells. The {sup 99}Tc concentration ranges from 3 to 10{sup -4} Bq/L in cavity waters and from 0.3 to 10{sup -4} Bq/L in satellite wells. Downstream migration is apparent for both radionuclides. However, it is affected by both retardation and initial distribution. In-situ {sup 99}Tc and {sup 129}I K{sub d}s calculated using rubble and water concentrations are 3 to 22 mL/g and 0 to 0.12 mL/g, respectively and are suggestive of mildly reducing conditions. {sup 129}I distribution in the melt glass, rubble and groundwater of the Chancellor test cavity is 28%, 24% and 48%, respectively; for {sup 99}Tc, it is 65%, 35% and 0.3%, respectively. Our partitioning estimates differ from those of underground tests in French Polynesia, implying that fission product …

The Yucca Mountain Performance Confirmation program consists of tests, monitoring activities, experiments, and analyses to evaluate the adequacy of assumptions, data, and analyses that form the basis of the conceptual and numerical models of flow and transport associated with a proposed radioactive waste repository at Yucca Mountain, Nevada. The Performance Confirmation program uses an eight-stage risk-informed, performance-based approach. Selection of the Performance Confirmation activities (a parameter and a test method) for inclusion in the Performance Confirmation program was done using a risk-informed performance-based decision analysis. The result of this analysis and review was a Performance Confirmation base portfolio that consists of 20 activities. The 20 Performance Confirmation activities include geologic, hydrologic, and construction/engineering testing. Several of the activities were initiated during site characterization and are ongoing. Others activities will commence during construction and/or post emplacement and will continue until repository closure.

The Yucca Mountain Performance Confirmation program consists of tests, monitoring activities, experiments, and analyses to evaluate the adequacy of assumptions, data, and analyses that form the basis of the conceptual and numerical models of flow and transport associated with a proposed radioactive waste repository at Yucca Mountain, Nevada. The Performance Confirmation program uses an eight-stage risk-informed, performance-based approach. Selection of the Performance Confirmation activities (a parameter and a test method) for inclusion in the Performance Confirmation program was done using a risk-informed performance-based decision analysis. The result of this analysis and review was a Performance Confirmation base portfolio that consists of 20 activities. The 20 Performance Confirmation activities include geologic, hydrologic, and construction/engineering testing. Several of the activities were initiated during site characterization and are ongoing. Others activities will commence during construction and/or post emplacement and will continue until repository closure.

As part of the effort to understand the flow and transport characteristics downgradient from the proposed high-level radioactive waste geologic repository at Yucca Mountain, Nevada, single- and cross-hole tracer tests were conducted from December 2004 through October 2005 in boreholes at the Nye County 22 well complex. The results were analyzed for transport properties using both numerical and analytical solutions of the governing advection dispersion equation. Preliminary results indicate effective flow porosity values ranging from 1.0 x 10{sup -2} for an individual flow path to 2.0 x 10{sup -1} for composite flow paths, longitudinal dispersivity ranging from 0.3 to 3 m, and a transverse horizontal dispersivity of 0.03 m. Individual flow paths identified from the cross-hole testing indicate some solute diffusion into the stagnant portion of the alluvial aquifer.

This document provides preplanning necessary to expeditiously remove any waste that may leak from the primary tank to the secondary tank for Hanford's 28 DSTs. The strategy is described, applicable emergency procedures are referenced, and transfer routes and pumping equipment for each tank are identified.