Rail-Cask Tests: Normal-Conditionsof- Transport Tests of Surrogate PWR Fuel Assemblies in an ENSA ENUN 32P Cask.

DOE Office of Scientific and Technical Information (OSTI.GOV)

McConnell, Paul E.; Ross, Steven; Grey, Carissa Ann

This report describes tests conducted using a full-size rail cask, the ENSA ENUN 32P, involving handling of the cask and transport of the cask via truck, ships, and rail. The purpose of the tests was to measure strains and accelerations on surrogate pressurized water reactor fuel rods when the fuel assemblies were subjected to Normal Conditions of Transport within the rail cask. In addition, accelerations were measured on the transport platform, the cask cradle, the cask, and the basket within the cask holding the assemblies. These tests were an international collaboration that included Equipos Nucleares S.A., Sandia National Laboratories, Pacificmore » Northwest National Laboratory, Coordinadora Internacional de Cargas S.A., the Transportation Technology Center, Inc., the Korea Radioactive Waste Agency, and the Korea Atomic Energy Research Institute. All test results in this report are PRELIMINARY – complete analyses of test data will be completed and reported in FY18. However, preliminarily: The strains were exceedingly low on the surrogate fuel rods during the rail-cask tests for all the transport and handling modes. The test results provide a compelling technical basis for the safe transport of spent fuel.« less

COMPILATION OF DISPOSABLE SOLID WASTE CASK EVALUATIONS

DOE Office of Scientific and Technical Information (OSTI.GOV)

THIELGES, J.R.; CHASTAIN, S.A.

The Disposable Solid Waste Cask (DSWC) is a shielded cask capable of transporting, storing, and disposing of six non-fuel core components or approximately 27 cubic feet of radioactive solid waste. Five existing DSWCs are candidates for use in storing and disposing of non-fuel core components and radioactive solid waste from the Interim Examination and Maintenance Cell, ultimately shipping them to the 200 West Area disposal site for burial. A series of inspections, studies, analyses, and modifications were performed to ensure that these casks can be used to safely ship solid waste. These inspections, studies, analyses, and modifications are summarized andmore » attached in this report. Visual inspection of the casks interiors provided information with respect to condition of the casks inner liners. Because water was allowed to enter the casks for varying lengths of time, condition of the cask liner pipe to bottom plate weld was of concern. Based on the visual inspection and a corrosion study, it was concluded that four of the five casks can be used from a corrosion standpoint. Only DSWC S/N-004 would need additional inspection and analysis to determine its usefulness. The five remaining DSWCs underwent some modification to prepare them for use. The existing cask lifting inserts were found to be corroded and deemed unusable. New lifting anchor bolts were installed to replace the existing anchors. Alternate lift lugs were fabricated for use with the new lifting anchor bolts. The cask tiedown frame was modified to facilitate adjustment of the cask tiedowns. As a result of the above mentioned inspections, studies, analysis, and modifications, four of the five existing casks can be used to store and transport waste from the Interim Examination and Maintenance Cell to the disposal site for burial. The fifth cask, DSWC S/N-004, would require further inspections before it could be used.« less

Multiple-Angle Muon Radiography of a Dry Storage Cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

Durham, J. Matthew; Guardincerri, Elena; Morris, Christopher

A partially loaded dry storage cask was imaged using cosmic ray muons. Since the cask is large relative to the size of the muon tracking detectors, the instruments were placed at nine different positions around the cask to record data covering the entire fuel basket. We show that this technique can detect the removal of a single fuel assembly from the center of the cask.

A task analysis of the shift from teacher instructions to self-instructions in performing an in-common task.

PubMed

Grote, I; Rosales, J; Baer, D M

1996-11-01

Three preschool children repeatedly did four kinds of sorts with a deck of stimulus cards: a difficult, untaught target sort and three other sorts considered analytic of self-instructing the target performance. The untaught target sort was to find in a deck of cards those matching what two sample cards had in common. Most preschool children must be taught to mediate this problem. The three other kinds of sorts taught skills involved in the target performance or its mediation. As correct self-instructive talk emerged in the target sorts, it was confirmed. The untaught target sorts were interspersed infrequently among the three alternating directly taught skill sorts, to see if accurate target sorts, and accurate self-instructive talk about the target sorts, would emerge as the three skill sorts were mastered. As all the sorts progressed, increasing accuracy was seen first in the skill sorts and then in the untaught target sorts. All three subjects showed subsequent generalization to new target sorts involving other stimulus sets. Correct spontaneous self-instructions about the target sorts increased from near zero at the beginning of the experiment to consistency at its end. Thus the three skill sorts appeared sufficient for the emergence of a self-instructed solution to the previously insoluble target performance.

Used Fuel Cask Identification through Neutron Profile

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rauch, Eric Benton

2015-11-20

Currently, most spent fuel is stored near reactors. An interim consolidated fuel storage facility would receive fuel from multiple sites and store it in casks on site for decades. For successful operation of such a facility there is need for a way to restore continuity of knowledge if lost as well as a method that will indicate state of fuel inside the cask. Used nuclear fuel is identifiable by its radiation emission, both gamma and neutron. Neutron emission from fission products, multiplication from remaining fissile material, and the unique distribution of both in each cask produce a unique neutron signature.more » If two signatures taken at different times do not match, either changes within the fuel content or misidentification of a cask occurred. It was found that identification of cask loadings works well through the profile of emitted neutrons in simulated real casks. Even casks with similar overall neutron emission or average counts around the circumference can be distinguished from each other by analyzing the profile. In conclusion, (1) identification of unaltered casks through neutron signature profile is viable; (2) collecting the profile provides insight to the condition and intactness of the fuel stored inside the cask; and (3) the signature profile is stable over time.« less

78 FR 78285 - List of Approved Spent Fuel Storage Casks: HI-STORM 100 Cask System; Amendment No. 9

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-26

...-2012-0052] RIN 3150-AJ12 List of Approved Spent Fuel Storage Casks: HI-STORM 100 Cask System; Amendment... document proposed to amend the NRC's spent fuel storage regulations by revising the Holtec International HI...

Performance testing and analyses of the VSC-17 ventilated concrete cask. Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

McKinnon, M.A.; Dodge, R.E.; Schmitt, R.C.

1992-05-01

This document details performance test which was conducted on a Pacific Sierra Nuclear VSC-17 ventilated concrete storage cask configured for pressurized-water reactor (PWR) spent fuel. The performance test consisted of loading the VSC-17 cask with 17 canisters of consolidated PWR spent fuel from Virginia Power`s Surry and Florida Power & Light Turkey Point reactors. Cask surface, concrete, air channel surfaces, and fuel canister guide tube temperatures were measured, as were cask surface gamma and neutron dose rates. Testing was performed with vacuum, nitrogen, and helium backfill environments in a vertical cask orientation. Data on spent fuel integrity were also obtained.

SCAN+

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kenneth Krebs, John Svoboda

2009-11-01

SCAN+ is a software application specifically designed to control the positioning of a gamma spectrometer by a two dimensional translation system above spent fuel bundles located in a sealed spent fuel cask. The gamma spectrometer collects gamma spectrum information for the purpose of spent fuel cask fuel loading verification. SCAN+ performs manual and automatic gamma spectrometer positioning functions as-well-as exercising control of the gamma spectrometer data acquisitioning functions. Cask configuration files are used to determine the positions of spent fuel bundles. Cask scanning files are used to determine the desired scan paths for scanning a spent fuel cask allowing formore » automatic unattended cask scanning that may take several hours.« less

Testing and COBRA-SFS analysis of the VSC-17 ventilated concrete, spent fuel storage cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

McKinnon, M.A.; Dodge, R.E.; Schmitt, R.C.

1992-04-01

A performance test of a Pacific Sierra Nuclear VSC-17 ventilated concrete storage cask loaded with 17 canisters of consolidated PWR spent fuel generating approximately 15 kW was conducted. The performance test included measuring the cask surface, concrete, air channel surface, and fuel temperatures, as well as cask surface gamma and neutron dose rates. Testing was performed using vacuum, nitrogen, and helium backfill environments. Pretest predictions of cask thermal performance were made using the COBRA-SFS computer code. Analysis results were within 15{degrees}C of measured peak fuel temperature. Peak fuel temperature for normal operation was 321{degrees}C. In general, the surface dose ratesmore » were less than 30 mrem/h on the side of the cask and 40 mrem/h on the top of the cask.« less

Axon-Axon Interactions Regulate Topographic Optic Tract Sorting via CYFIP2-Dependent WAVE Complex Function.

PubMed

Cioni, Jean-Michel; Wong, Hovy Ho-Wai; Bressan, Dario; Kodama, Lay; Harris, William A; Holt, Christine E

2018-03-07

The axons of retinal ganglion cells (RGCs) are topographically sorted before they arrive at the optic tectum. This pre-target sorting, typical of axon tracts throughout the brain, is poorly understood. Here, we show that cytoplasmic FMR1-interacting proteins (CYFIPs) fulfill non-redundant functions in RGCs, with CYFIP1 mediating axon growth and CYFIP2 specifically involved in axon sorting. We find that CYFIP2 mediates homotypic and heterotypic contact-triggered fasciculation and repulsion responses between dorsal and ventral axons. CYFIP2 associates with transporting ribonucleoprotein particles in axons and regulates translation. Axon-axon contact stimulates CYFIP2 to move into growth cones where it joins the actin nucleating WAVE regulatory complex (WRC) in the periphery and regulates actin remodeling and filopodial dynamics. CYFIP2's function in axon sorting is mediated by its binding to the WRC but not its translational regulation. Together, these findings uncover CYFIP2 as a key regulatory link between axon-axon interactions, filopodial dynamics, and optic tract sorting. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Thermal analyses of the IF-300 shipping cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

Meier, J.K.

1978-07-01

In order to supply temperature data for structural testing and analysis of shipping casks, a series of thermal analyses using the TRUMP thermal analyzer program were performed on the GE IF-300 spent fuel shipping cask. Major conclusions of the analyses are: (1) Under normal cooling conditions and a cask heat load of 262,000 BTU/h, the seal area of the cask will be roughly 100/sup 0/C (180/sup 0/F) above the ambient surroundings. (2) Under these same conditions the uranium shield at the midpoint of the cask will be between 69/sup 0/C (125/sup 0/F) and 92/sup 0/C (166/sup 0/F) above the ambientmore » surroundings. (3) Significant thermal gradients are not likely to develop between the head studs and the surrounding metal. (4) A representative time constant for the cask as a whole is on the order of one day.« less

76 FR 2277 - List of Approved Spent Fuel Storage Casks: NUHOMS® HD System Revision 1

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-13

... Fuel Storage Casks: NUHOMS[supreg] HD System Revision 1 AGENCY: Nuclear Regulatory Commission. ACTION... amend its spent fuel storage cask regulations by revising the Transnuclear, Inc. (TN) NUHOMS[supreg] HD System listing within the ``List of Approved Spent Fuel Storage Casks'' to include Amendment No. 1 to...

Development of New Transportation/Storage Cask System for Use by DOE Russian Research Reactor Fuel Return Program

DOE Office of Scientific and Technical Information (OSTI.GOV)

Michael Tyacke; Frantisek Svitak; Jiri Rychecky

2010-04-01

The United States, the Russian Federation, and the International Atomic Energy Agency (IAEA) have been working together on a program called the Russian Research Reactor Fuel Return (RRRFR) Program. The purpose of this program is to return Soviet or Russian supplied high-enriched uranium (HEU) fuel currently stored at Russian-designed research reactors throughout the world to Russia. To accommodate transport of the HEU spent nuclear fuel (SNF), a new large-capacity transport/storage cask system was specially designed for handling and operations under the unique conditions for these research reactor facilities. This new cask system is named the ŠKODA VPVR/M cask. The design,more » licensing, testing, and delivery of this new cask system are the results of a significant international cooperative effort by several countries and involved numerous private and governmental organizations. This paper contains the following sections: (1) Introduction/Background; (2) VPVR/M Cask Description; (3) Ancillary Equipment, (4) Cask Licensing; (5) Cask Demonstration and Operations; (6) IAEA Procurement, Quality Assurance Inspections, Fabrication, and Delivery; and, (7) Summary and Conclusions.« less

Development of a New Transportation/Storage Cask System for Use by the DOE Russian Research Reactor Fuel Return Program

DOE Office of Scientific and Technical Information (OSTI.GOV)

Michael J. Tyacke; Frantisek Svitak; Jiri Rychecky

2007-10-01

The United States, the Russian Federation, and the International Atomic Energy Agency (IAEA) have been working together on a program called the Russian Research Reactor Fuel Return (RRRFR) Program. The purpose of this program is to return Soviet or Russian-supplied high-enriched uranium (HEU) fuel, currently stored at Russian-designed research reactors throughout the world, to Russia. To accommodate transport of the HEU spent nuclear fuel (SNF), a new large-capacity transport/storage cask system was specially designed for handling and operations under the unique conditions at these research reactor facilities. This new cask system is named the ŠKODA VPVR/M cask. The design, licensing,more » testing, and delivery of this new cask system result from a significant international cooperative effort by several countries and involved numerous private and governmental organizations. This paper contains the following sections: 1) Introduction; 2) VPVR/M Cask Description; 3) Ancillary Equipment, 4) Cask Licensing; 5) Cask Demonstration and Operations; 6) IAEA Procurement, Quality Assurance Inspections, Fabrication, and Delivery; and, 7) Conclusions.« less

A preliminary evaluation of the ability of from-reactor casks to geometrically accommodate commercial LWR spent nuclear fuel

DOE Office of Scientific and Technical Information (OSTI.GOV)

Andress, D.; Joy, D.S.; McLeod, N.B.

The Department of Energy has sponsored a number of cask design efforts to define several transportation casks to accommodate the various assemblies expected to be accepted by the Federal Waste Management System. At this time, three preliminary cask designs have been selected for the final design--the GA-4 and GA-9 truck casks and the BR-100 rail cask. In total, this assessment indicates that the current Initiative I cask designs can be expected to dimensionally accommodate 100% of the PWR fuel assemblies (other than the extra-long South Texas Fuel) with control elements removed, and >90% of the assemblies having the control elementsmore » as an integral part of the fuel assembly. For BWR assemblies, >99% of the assemblies can be accommodated with fuel channels removed. This paper summarizes preliminary results of one part of that evaluation related to the ability of the From-Reactor Initiative I casks to accommodate the physical and radiological characteristics of the Spent Nuclear Fuel projected to be accepted into the Federal Waste Management System. 3 refs., 5 tabs.« less

Nondestructive Evaluation of the VSC-17 Cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sheryl Morton; Al Carlson; Cecilia Hoffman

2006-01-01

In 2003, representatives from the Central Research Institute of Electric Power Industry (CRIEPI) requested development of a project with the objective of determining the performance of a concrete spent nuclear fuel storage cask. Radiation and environmental effects may cause chemical alteration of the concrete that could result in excessive cracking, spalling, and loss of compressive strength. The Idaho National Laboratory (INL) project team and CRIEPI representatives identified the Ventilated Storage Cask (VSC 17) spent nuclear fuel storage cask, originally located at the INL Test Area North, as a candidate to study cask performance because it had been used to storemore » fuel as part of a dry cask storage demonstration project for over 15 years. The project involved investigating the properties of the concrete shield. INL performed a survey of the cask in the summers of 2003 and 2004. The INL team met with the CRIEPI representatives in December of 2004 to discuss the next steps. As a result of that meeting, CRIEPI requested that in the summer 2005 INL perform additional surveys on the VSC 17 cask with participation of CRIEPI scientists. This document summarizes the evaluation methods used on the VSC 17 to evaluate the cask for compressive strength, concrete cracking, concrete thickness, and temperature distribution.« less

78 FR 22411 - List of Approved Spent Fuel Storage Casks: HI-STORM 100, Amendment No. 8; Corrections

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-16

... Fuel Storage Casks: HI-STORM 100, Amendment No. 8; Corrections AGENCY: Nuclear Regulatory Commission... revising the Holtec International, Inc. (Holtec) HI-STORM 100 Cask System listing within the ``List of... the Holtec HI-STORM 100 Cask System, Amendment No. 8. The purpose of this document is to provide...

75 FR 27463 - List of Approved Spent Fuel Storage Casks: NUHOMS® HD System Revision 1; Correction

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-17

... Fuel Storage Casks: NUHOMS[supreg] HD System Revision 1; Correction AGENCY: Nuclear Regulatory... fuel storage casks to add revision 1 to the NUHOMS HD spent fuel storage cask system. This action is... Federal Register on May 7, 2010 (75 FR 25120), that proposes to amend the regulations that govern storage...

Results for the Aboveground Configuration of the Boiling Water Reactor Dry Cask Simulator

DOE Office of Scientific and Technical Information (OSTI.GOV)

Durbin, Samuel G.; Lindgren, Eric R.

The thermal performance of commercial nuclear spent fuel dry storage casks is evaluated through detailed numerical analysis. These modeling efforts are completed by the vendor to demonstrate performance and regulatory compliance. The calculations are then independently verified by the Nuclear Regulatory Commission (NRC). Carefully measured data sets generated from testing of full-sized casks or smaller cask analogs are widely recognized as vital for validating these models. Recent advances in dry storage cask designs have significantly increased the maximum thermal load allowed in a cask, in part by increasing the efficiency of internal conduction pathways, and also by increasing the internalmore » convection through greater canister helium pressure. These same canistered cask systems rely on ventilation between the canister and the overpack to convect heat away from the canister to the environment for both above- and below-ground configurations. While several testing programs have been previously conducted, these earlier validation attempts did not capture the effects of elevated helium pressures or accurately portray the external convection of above-ground and below-ground canistered dry cask systems. The purpose of the current investigation was to produce data sets that can be used to test the validity of the assumptions associated with the calculations used to determine steady-state cladding temperatures in modern dry casks that utilize elevated helium pressure in the sealed canister in an above-ground configuration.« less

76 FR 33121 - List of Approved Spent Fuel Storage Casks: HI-STORM Flood/Wind Addition

Federal Register 2010, 2011, 2012, 2013, 2014

2011-06-08

... Storage Casks: HI-STORM Flood/Wind Addition AGENCY: Nuclear Regulatory Commission. ACTION: Direct final... regulations to add the Holtec HI-STORM Flood/Wind cask system to the ``List of Approved Spent Fuel Storage... Title 10 of the Code of Federal Regulations Section 72.214 to add the Holtec HI- STORM Flood/Wind cask...

Feasibility Study For Use Of Commercial Cask Vendor Dry Transfer Systems To Unload Used Fuel Assemblies In L-Area

DOE Office of Scientific and Technical Information (OSTI.GOV)

Krementz, Dan; Rose, David; Dunsmuir, Mike

2014-02-06

The purpose of this study is to determine whether a commercial dry transfer system (DTS) could be used for loading or unloading used nuclear fuel (UNF) in L-Basin and to determine if a DTS pool adapter could be made for L-Basin Transfer Pit #2 that could accommodate a variety of DTS casks and fuel baskets or canisters up to 24” diameter.[1, 2] This study outlines the technical feasibility of accommodating different vendor dry transfer systems in the L-Basin Transfer Bay with a general work scope. It identifies equipment needing development, facility modifications, and describes the needed analyses and calculations. Aftermore » reviewing the L-Basin Transfer Bay area layout and information on the only DTS system currently in use for the Nuclear Assurance Corporation Legal Weight Truck cask (NAC LWT), the authors conclude that use of a dry transfer cask is feasible. AREVA was contacted and acknowledged that they currently do not have a design for a dry transfer cask for their new Transnuclear Long Cask (TN-LC) cask. Nonetheless, this study accounted for a potential future DTS from AREVA to handle fuel baskets up to 18” in diameter. Due to the layout of the Transfer Bay, it was determined that a DTS cask pool adapter designed specifically for spanning Pit #2 and placed just north of the 70 Ton Cask lid lifting superstructure would be needed. The proposed pool adapter could be used to transition a fuel basket up to 24” in diameter and ~11 feet long from a dry transfer cask to the basin. The 18” and 24” applications of the pool adapter are pending vendor development of dry transfer casks that accommodate these diameters. Once a fuel basket has been lowered into Pit #2 through a pool adapter, a basket cart could be used to move the basket out from under the pool adapter for access by the 5 Ton Crane. The cost to install a dry transfer cask handling system in L-Area capable of handling multiple vendor provided transport and dry transfer casks and baskets with different diameters and lengths would likely be on the same order of magnitude as the Basin Modifications project. The cost of a DTS capability is affected by the number of design variations of different vendor transport and dry transfer casks to be considered for design input. Some costs would be incurred for each vendor DTS to be handled. For example, separate analyses would be needed for each dry transfer cask type such as criticality, shielding, dropping a dry transfer cask and basket, handling and auxiliary equipment, procedures, operator training, readiness assessments, and operational readiness reviews. A DTS handling capability in L-Area could serve as a backup to the Shielded Transfer System (STS) for unloading long casks and could support potential future missions such as the Idaho National Laboratory (INL) Exchange or transferring UNF from wet to dry storage.« less

Early thermal testing of type B radioactive material packages in USA to environments beyond regulatory package thermal test standards

DOE PAGES

Yoshimura, H. R.; Pope, R. B.; Kubo, M.

2007-06-01

Three separate fire test programmes exposing casks beyond the regulatory thermal test requirements were performed by Sandia National Laboratories during the late 1970s and mid 1980s. The results of these test programmes can be used to assist in addressing the adequacy of the regulatory thermal test of fully engulfing exposure at 800°C for 30 min and how that test might relate to real accident thermal environments. The test programmes were undertaken on obsolete and new casks on behalf of the US Department of Energy (DOE), the US Department of Transportation (DOT) and the Japanese Power Reactor and Nuclear Fuel Developmentmore » Corporation (PNC), currently known as the Japan Atomic Energy Agency. Two of the tests involved exposure of casks in damaged transport vehicles to fully engulfing fires for 72–125 min, and the other test involved four exposures of a cask to torch environments for 30 min. Much of the original documentation regarding these tests and their results is no longer readily available. The documents relating to these tests have been surveyed; this paper presents summaries from this survey of the tests and their results. Specifically, for the pool fire exposures, the temperatures measured in the flames of both exceeded the flame temperature required by the Transport Regulations; yet an obsolete 67 t cask endured 90 min of exposure before evidence of failure was detected, and a new cask endured the 72 min exposure while retaining its containment integrity. For the exposure of a modified obsolete cask to four different torch environments, the integrity of the cask was retained and the relative temperature increases within the cask were well within acceptable limits and well below the values that could be expected if the cask was exposed to the regulatory thermal test. In this paper, a review of these three thermal test programmes, establishes that the two older cask designs and one new cask design have the ability to survive environments that were different from (the torch environments) or more severe than the environment specified by the existing thermal test requirement in the Transport Regulations. Finally, these results can be extrapolated to apply to modern casks that generally have more robust designs as well as better quality assurance applied during the manufacturing process.« less

CASK and CaMKII function in Drosophila memory

PubMed Central

Malik, Bilal R.; Hodge, James J. L.

2014-01-01

Calcium (Ca2+) and Calmodulin (CaM)-dependent serine/threonine kinase II (CaMKII) plays a central role in synaptic plasticity and memory due to its ability to phosphorylate itself and regulate its own kinase activity. Autophosphorylation at threonine 287 (T287) switches CaMKII to a Ca2+ independent and constitutively active state replicated by overexpression of a phosphomimetic CaMKII-T287D transgene or blocked by expression of a T287A transgene. A second pair of sites, T306 T307 in the CaM binding region once autophosphorylated, prevents CaM binding and inactivates the kinase during synaptic plasticity and memory, and can be blocked by a TT306/7AA transgene. Recently the synaptic scaffolding molecule called CASK (Ca2+/CaM-associated serine kinase) has been shown to control both sets of CaMKII autophosphorylation events during neuronal growth, Ca2+ signaling and memory in Drosophila. Deletion of either full length CASK or just its CaMK-like and L27 domains removed middle-term memory (MTM) and long-term memory (LTM), with CASK function in the α′/ß′ mushroom body neurons being required for memory. In a similar manner directly changing the levels of CaMKII autophosphorylation (T287D, T287A, or TT306/7AA) in the α′/ß′ neurons also removed MTM and LTM. In the CASK null mutant expression of either the Drosophila or human CASK transgene in the α′/ß′ neurons was found to completely rescue memory, confirming that CASK signaling in α′/β′ neurons is necessary and sufficient for Drosophila memory formation and that the neuronal function of CASK is conserved between Drosophila and human. Expression of human CASK in Drosophila also rescued the effect of CASK deletion on the activity state of CaMKII, suggesting that human CASK may also regulate CaMKII autophosphorylation. Mutations in human CASK have recently been shown to result in intellectual disability and neurological defects suggesting a role in plasticity and learning possibly via regulation of CaMKII autophosphorylation. PMID:25009461

CASK and CaMKII function in the mushroom body α'/β' neurons during Drosophila memory formation.

PubMed

Malik, Bilal R; Gillespie, John Michael; Hodge, James J L

2013-01-01

Ca(2+)/CaM serine/threonine kinase II (CaMKII) is a central molecule in mechanisms of synaptic plasticity and memory. A vital feature of CaMKII in plasticity is its ability to switch to a calcium (Ca(2+)) independent constitutively active state after autophosphorylation at threonine 287 (T287). A second pair of sites, T306 T307 in the calmodulin (CaM) binding region once autophosphorylated, prevent subsequent CaM binding and inactivates the kinase during synaptic plasticity and memory. Recently a synaptic molecule called Ca(2+)/CaM-dependent serine protein kinase (CASK) has been shown to control both sets of CaMKII autophosphorylation events and hence is well poised to be a key regulator of memory. We show deletion of full length CASK or just its CaMK-like and L27 domains disrupts middle-term memory (MTM) and long-term memory (LTM), with CASK function in the α'/β' subset of mushroom body neurons being required for memory. Likewise directly changing the levels of CaMKII autophosphorylation in these neurons removed MTM and LTM. The requirement of CASK and CaMKII autophosphorylation was not developmental as their manipulation just in the adult α'/β' neurons was sufficient to remove memory. Overexpression of CASK or CaMKII in the α'/β' neurons also occluded MTM and LTM. Overexpression of either Drosophila or human CASK in the α'/β' neurons of the CASK mutant completely rescued memory, confirming that CASK signaling in α'/β' neurons is necessary and sufficient for Drosophila memory formation and that the neuronal function of CASK is conserved between Drosophila and human. At the cellular level CaMKII overexpression in the α'/β' neurons increased activity dependent Ca(2+) responses while reduction of CaMKII decreased it. Likewise reducing CASK or directly expressing a phosphomimetic CaMKII T287D transgene in the α'/β' similarly decreased Ca(2+) signaling. Our results are consistent with CASK regulating CaMKII autophosphorylation in a pathway required for memory formation that involves activity dependent changes in Ca(2+) signaling in the α'/β' neurons.

Status update of the BWR cask simulator

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lindgren, Eric R.; Durbin, Samuel G.

2015-09-01

The performance of commercial nuclear spent fuel dry storage casks are typically evaluated through detailed numerical analysis of the system's thermal performance. These modeling efforts are performed by the vendor to demonstrate the performance and regulatory compliance and are independently verified by the Nuclear Regulatory Commission (NRC). Carefully measured data sets generated from testing of full sized casks or smaller cask analogs are widely recognized as vital for validating these models. Numerous studies have been previously conducted. Recent advances in dry storage cask designs have moved the storage location from above ground to below ground and significantly increased the maximummore » thermal load allowed in a cask in part by increasing the canister helium pressure. Previous cask performance validation testing did not capture these parameters. The purpose of the investigation described in this report is to produce a data set that can be used to test the validity of the assumptions associated with the calculations presently used to determine steady-state cladding temperatures in modern dry casks. These modern cask designs utilize elevated helium pressure in the sealed canister or are intended for subsurface storage. The BWR cask simulator (BCS) has been designed in detail for both the above ground and below ground venting configurations. The pressure vessel representing the canister has been designed, fabricated, and pressure tested for a maximum allowable pressure (MAWP) rating of 24 bar at 400 C. An existing electrically heated but otherwise prototypic BWR Incoloy-clad test assembly is being deployed inside of a representative storage basket and cylindrical pressure vessel that represents the canister. The symmetric single assembly geometry with well-controlled boundary conditions simplifies interpretation of results. Various configurations of outer concentric ducting will be used to mimic conditions for above and below ground storage configurations of vertical, dry cask systems with canisters. Radial and axial temperature profiles will be measured for a wide range of decay power and helium cask pressures. Of particular interest is the evaluation of the effect of increased helium pressure on heat load and the effect of simulated wind on a simplified below ground vent configuration.« less

Viability of Existing INL Facilities for Dry Storage Cask Handling

DOE Office of Scientific and Technical Information (OSTI.GOV)

Randy Bohachek; Charles Park; Bruce Wallace

2013-04-01

This report evaluates existing capabilities at the INL to determine if a practical and cost effective method could be developed for opening and handling full-sized dry storage casks. The Idaho Nuclear Technology and Engineering Center (INTEC) CPP-603, Irradiated Spent Fuel Storage Facility, provides the infrastructure to support handling and examining casks and their contents. Based on a reasonable set of assumptions, it is possible to receive, open, inspect, remove samples, close, and reseal large bolted-lid dry storage casks at the INL. The capability can also be used to open and inspect casks that were last examined at the TAN Hotmore » Shop over ten years ago. The Castor V/21 and REA-2023 casks can provide additional confirmatory information regarding the extended performance of low-burnup (<45 GWD/MTU) used nuclear fuel. Once a dry storage cask is opened inside CPP-603, used fuel retrieved from the cask can be packaged in a shipping cask, and sent to a laboratory for testing. Testing at the INL’s Materials and Fuels Complex (MFC) can occur starting with shipment of samples from CPP-603 over an on-site road, avoiding the need to use public highways. This reduces cost and reduces the risk to the public. The full suite of characterization methods needed to establish the condition of the fuel exists and MFC. Many other testing capabilities also exist at MFC, but when those capabilities are not adequate, samples can be prepared and shipped to other laboratories for testing. This report discusses how the casks would be handled, what work needs to be done to ready the facilities/capabilities, and what the work will cost.« less

Viability of Existing INL Facilities for Dry Storage Cask Handling

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bohachek, Randy; Wallace, Bruce; Winston, Phil

2013-04-30

This report evaluates existing capabilities at the INL to determine if a practical and cost effective method could be developed for opening and handling full-sized dry storage casks. The Idaho Nuclear Technology and Engineering Center (INTEC) CPP-603, Irradiated Spent Fuel Storage Facility, provides the infrastructure to support handling and examining casks and their contents. Based on a reasonable set of assumptions, it is possible to receive, open, inspect, remove samples, close, and reseal large bolted-lid dry storage casks at the INL. The capability can also be used to open and inspect casks that were last examined at the TAN Hotmore » Shop over ten years ago. The Castor V/21 and REA-2023 casks can provide additional confirmatory information regarding the extended performance of low-burnup (<45 GWD/MTU) used nuclear fuel. Once a dry storage cask is opened inside CPP-603, used fuel retrieved from the cask can be packaged in a shipping cask, and sent to a laboratory for testing. Testing at the INL’s Materials and Fuels Complex (MFC) can occur starting with shipment of samples from CPP-603 over an on-site road, avoiding the need to use public highways. This reduces cost and reduces the risk to the public. The full suite of characterization methods needed to establish the condition of the fuel exists and MFC. Many other testing capabilities also exist at MFC, but when those capabilities are not adequate, samples can be prepared and shipped to other laboratories for testing. This report discusses how the casks would be handled, what work needs to be done to ready the facilities/capabilities, and what the work will cost.« less

Genetics Home Reference: CASK-related intellectual disability

MedlinePlus

... XL-ID with or without nystagmus (rapid, involuntary eye movements) is a milder form of CASK -related intellectual ... to promote development of the nerves that control eye movement (the oculomotor neural network). Mutations in the CASK ...

Test Plan for Cask Identification Detector

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rauch, Eric Benton

2016-09-29

This document serves to outline the testing of a Used Fuel Cask Identification Detector (CID) currently being designed under the DOE-NE MPACT Campaign. A bench-scale prototype detector will be constructed and tested using surrogate neutron sources. The testing will serve to inform the design of the full detector that is to be used as a way of fingerprinting used fuel storage casks based on the neutron signature produced by the used fuel inside the cask.

Development of a conditioning system for the dual-purpose transport and storage cask for spent nuclear fuel from decommissioned Russian submarines

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dyer, R.S.; Barnes, E.; Snipes, R.L.

2007-07-01

Russia, stores large quantities of spent nuclear fuel (SNF) from submarine and ice-breaker nuclear powered naval vessels. This high-level radioactive material presents a significant threat to the Arctic and marine environments. Much of the SNF from decommissioned Russian nuclear submarines is stored either onboard the submarines or in floating storage vessels in Northwest and Far East Russia. Some of the SNF is damaged, stored in an unstable condition, or of a type that cannot currently be reprocessed. In many cases, the existing Russian transport infrastructure and reprocessing facilities cannot meet the requirements for moving and reprocessing all of this fuelmore » from remote locations. Additional transport and storage options are required. Some of the existing storage facilities being used in Russia do not meet health and safety and physical security requirements. The U.S. has assisted Russia in the development of a new dual-purpose metal-concrete transport and storage cask (TUK-108/1) for their military SNF and assisted them in building several new facilities for off-loading submarine SNF and storing these TUK-108/1 casks. These efforts have reduced the technical, ecological, and security challenges for removal, handling, interim storage, and shipment of this submarine fuel. Currently, Russian licensing limits the storage period of the TUK-108/1 casks to no more than two years before the fuel must be shipped for reprocessing. In order to extend this licensed storage period, a system is required to condition the casks by removing residual water and creating an inert storage environment by backfilling the internal canisters with a noble gas such as argon. The U.S. has assisted Russia in the development of a mobile cask conditioning system for the TUK-108/1 cask. This new conditioning system allows the TUK 108/1 casks to be stored for up to five years after which the license may be considered for renewal for an additional five years or the fuel will be shipped to 'Mayak' for reprocessing. The U.S. Environmental Protection Agency (EPA), in cooperation with the U.S. DOD Office of Cooperative Threat Reduction (CTR), and the DOE's ORNL, along with the Norwegian Defense Research Establishment, worked closely with the Ministry of Defense and the Ministry of Atomic Energy of the Russian Federation (RF) to develop an improved integrated management system for interim storage of military SNF in Russia. The initial Project activities included: (1) development of a prototype dual-purpose, metal-concrete 40-ton cask for both the transport and interim storage of RF SNF, and (2) development of the first transshipment/interim storage facility for these casks in Murmansk. The U.S. has continued support to the project by assisting the RF with the development of the first mobile system that provides internal conditioning for the TUK-108/1 casks to allow them to be stored for longer than the current licensing period of two years. Development of the prototype TUK-108/1 cask was completed in December 2000 under the Arctic Military Environmental Cooperation (AMEC) Program. This was the first metal-concrete cask developed, licensed, and produced in the RF for both the transportation and storage of SNF from decommissioned submarines. These casks are currently being serially produced in NW Russia and 108 casks have been produced to date. Russia is using these casks for the transport and interim storage of military SNF from decommissioned nuclear submarines at naval installations in the Arctic and Far East in conformance with the Strategic Arms Reduction Treaty (START II). The design, construction, and commissioning of the first transshipment/interim storage facility in the RF was completed and ready for full operation in September 2003. Because of the RF government reorganization and changing regulations for spent fuel storage facilities, the storage facility at Murmansk was not fully licensed for operation until December 2005. The RF has reported that the facility is now fully operational. The TUK-108/1 SNF transport and storage casks were designed to have a 50-year storage life. Current RF practice is not to condition the submarine SNF or cask during the cask loading. Current RF regulations allow up to 4 mm of residual water (up to 3.2 liters) to remain in the casks. It has been determined that allowing this amount of residual water to remain untreated for a period longer than two years can produce hydrogen gas through hydrolysis which will increase the risk of explosion and could cause some corrosion of internal components. A solution to this problem was to develop and utilize a cask conditioning system to remove the residual water and create an inert storage environment in the cask by back-filling the internal cask cavity with an inert gas, such as helium or argon. This system is compatible with the existing TUK-108/1 design and is mobile for use at multiple submarine dismantlement sites. The RF has required that this cask conditioning system be tested and commissioned at the 'Zvezda' Shipyard in the Far East near Vladivostok, one of the major RF submarine fuel off loading and storage facilities. Currently, the fuel cannot be transferred to 'Mayak' for reprocessing until the completion of the 20 km railroad connector between 'Zvezda' and the main rail line to 'Mayak'. The cask conditioning system will allow extension of the currently-stored casks for an additional three years, at which time the rail connector line should be completed. The current license to store these casks at 'Zvezda' was scheduled to expire on 31 Dec 2006. Without the cask-conditioning system, the license could not be extended, no more fuel could be off-loaded from the decommissioned submarines, and the START objectives could not be met at 'Zvezda'. Completion of this cask conditioning system has removed a significant bottleneck for the completion of the Russian submarine decommissioning program under the START II Agreement. (authors)« less

CASK and CaMKII function in the mushroom body α′/β′ neurons during Drosophila memory formation

PubMed Central

Malik, Bilal R.; Gillespie, John Michael; Hodge, James J. L.

2013-01-01

Ca2+/CaM serine/threonine kinase II (CaMKII) is a central molecule in mechanisms of synaptic plasticity and memory. A vital feature of CaMKII in plasticity is its ability to switch to a calcium (Ca2+) independent constitutively active state after autophosphorylation at threonine 287 (T287). A second pair of sites, T306 T307 in the calmodulin (CaM) binding region once autophosphorylated, prevent subsequent CaM binding and inactivates the kinase during synaptic plasticity and memory. Recently a synaptic molecule called Ca2+/CaM-dependent serine protein kinase (CASK) has been shown to control both sets of CaMKII autophosphorylation events and hence is well poised to be a key regulator of memory. We show deletion of full length CASK or just its CaMK-like and L27 domains disrupts middle-term memory (MTM) and long-term memory (LTM), with CASK function in the α′/β′ subset of mushroom body neurons being required for memory. Likewise directly changing the levels of CaMKII autophosphorylation in these neurons removed MTM and LTM. The requirement of CASK and CaMKII autophosphorylation was not developmental as their manipulation just in the adult α′/β′ neurons was sufficient to remove memory. Overexpression of CASK or CaMKII in the α′/β′ neurons also occluded MTM and LTM. Overexpression of either Drosophila or human CASK in the α′/β′ neurons of the CASK mutant completely rescued memory, confirming that CASK signaling in α′/β′ neurons is necessary and sufficient for Drosophila memory formation and that the neuronal function of CASK is conserved between Drosophila and human. At the cellular level CaMKII overexpression in the α′/β′ neurons increased activity dependent Ca2+ responses while reduction of CaMKII decreased it. Likewise reducing CASK or directly expressing a phosphomimetic CaMKII T287D transgene in the α′/β′ similarly decreased Ca2+ signaling. Our results are consistent with CASK regulating CaMKII autophosphorylation in a pathway required for memory formation that involves activity dependent changes in Ca2+ signaling in the α′/β′ neurons. PMID:23543616

Signatures of Extended Storage of Used Nuclear Fuel in Casks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rauch, Eric Benton

2016-09-28

As the amount of used nuclear fuel continues to grow, more and more used nuclear fuel will be transferred to storage casks. A consolidated storage facility is currently in the planning stages for storing these casks, where at least 10,000 MTHM of fuel will be stored. This site will have potentially thousands of casks once it is operational. A facility this large presents new safeguards and nuclear material accounting concerns. A new signature based on the distribution of neutron sources and multiplication within casks was part of the Department of Energy Office of Nuclear Energy’s Material Protection, Account and Controlmore » Technologies (MPACT) campaign. Under this project we looked at fingerprinting each cask's neutron signature. Each cask has a unique set of fuel, with a unique spread of initial enrichment, burnup, cooling time, and power history. The unique set of fuel creates a unique signature of neutron intensity based on the arrangement of the assemblies. The unique arrangement of neutron sources and multiplication produces a reliable and unique identification of the cask that has been shown to be relatively constant over long time periods. The work presented here could be used to restore from a loss of continuity of knowledge at the storage site. This presentation will show the steps used to simulate and form this signature from the start of the effort through its conclusion in September 2016.« less

Adapting Dry Cask Storage for Aging at a Geologic Repository

DOE Office of Scientific and Technical Information (OSTI.GOV)

C. Sanders; D. Kimball

2005-08-02

A Spent Nuclear Fuel (SNF) Aging System is a crucial part of operations at the proposed Yucca Mountain repository in the United States. Incoming commercial SNF that does not meet thermal limits for emplacement will be aged on outdoor pads. U.S. Department of Energy SNF will also be managed using the Aging System. Proposed site-specific designs for the Aging System are closely based upon designs for existing dry cask storage (DCS) systems. This paper evaluates the applicability of existing DCS systems for use in the SNF Aging System at Yucca Mountain. The most important difference between existing DCS facilities andmore » the Yucca Mountain facility is the required capacity. Existing DCS facilities typically have less than 50 casks. The current design for the aging pad at Yucca Mountain calls for a capacity of over 2,000 casks (20,000 MTHM) [1]. This unprecedented number of casks poses some unique problems. The response of DCS systems to off-normal and accident conditions needs to be re-evaluated for multiple storage casks. Dose calculations become more complicated, since doses from multiple or very long arrays of casks can dramatically increase the total boundary dose. For occupational doses, the geometry of the cask arrays and the order of loading casks must be carefully considered in order to meet ALARA goals during cask retrieval. Due to the large area of the aging pad, skyshine must also be included when calculating public and worker doses. The expected length of aging will also necessitate some design adjustments. Under 10 CFR 72.236, DCS systems are initially certified for a period of 20 years [2]. Although the Yucca Mountain facility is not intended to be a storage facility under 10 CFR 72, the operational life of the SNF Aging System is 50 years [1]. Any cask system selected for use in aging will have to be qualified to this design lifetime. These considerations are examined, and a summary is provided of the adaptations that must be made in order to use DCS technologies successfully at a geologic repository.« less

Full-Scale Cask Testing and Public Acceptance of Spent Nuclear Fuel Shipments - 12254

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dilger, Fred; Halstead, Robert J.; Ballard, James D.

Full-scale physical testing of spent fuel shipping casks has been proposed by the National Academy of Sciences (NAS) 2006 report on spent nuclear fuel transportation, and by the Presidential Blue Ribbon Commission (BRC) on America's Nuclear Future 2011 draft report. The U.S. Nuclear Regulatory Commission (NRC) in 2005 proposed full-scale testing of a rail cask, and considered 'regulatory limits' testing of both rail and truck casks (SRM SECY-05-0051). The recent U.S. Department of Energy (DOE) cancellation of the Yucca Mountain project, NRC evaluation of extended spent fuel storage (possibly beyond 60-120 years) before transportation, nuclear industry adoption of very largemore » dual-purpose canisters for spent fuel storage and transport, and the deliberations of the BRC, will fundamentally change assumptions about the future spent fuel transportation system, and reopen the debate over shipping cask performance in severe accidents and acts of sabotage. This paper examines possible approaches to full-scale testing for enhancing public confidence in risk analyses, perception of risk, and acceptance of spent fuel shipments. The paper reviews the literature on public perception of spent nuclear fuel and nuclear waste transportation risks. We review and summarize opinion surveys sponsored by the State of Nevada over the past two decades, which show consistent patterns of concern among Nevada residents about health and safety impacts, and socioeconomic impacts such as reduced property values along likely transportation routes. We also review and summarize the large body of public opinion survey research on transportation concerns at regional and national levels. The paper reviews three past cask testing programs, the way in which these cask testing program results were portrayed in films and videos, and examines public and official responses to these three programs: the 1970's impact and fire testing of spent fuel truck casks at Sandia National Laboratories, the 1980's regulatory and demonstration testing of MAGNOX fuel flasks in the United Kingdom (the CEGB 'Operation Smash Hit' tests), and the 1980's regulatory drop and fire tests conducted on the TRUPACT II containers used for transuranic waste shipments to the Waste Isolation Pilot Plant in New Mexico. The primary focus of the paper is a detailed evaluation of the cask testing programs proposed by the NRC in its decision implementing staff recommendations based on the Package Performance Study, and by the State of Nevada recommendations based on previous work by Audin, Resnikoff, Dilger, Halstead, and Greiner. The NRC approach is based on demonstration impact testing (locomotive strike) of a large rail cask, either the TAD cask proposed by DOE for spent fuel shipments to Yucca Mountain, or a similar currently licensed dual-purpose cask. The NRC program might also be expanded to include fire testing of a legal-weight truck cask. The Nevada approach calls for a minimum of two tests: regulatory testing (impact, fire, puncture, immersion) of a rail cask, and extra-regulatory fire testing of a legal-weight truck cask, based on the cask performance modeling work by Greiner. The paper concludes with a discussion of key procedural elements - test costs and funding sources, development of testing protocols, selection of testing facilities, and test peer review - and various methods of communicating the test results to a broad range of stakeholder audiences. (authors)« less

CASK interacts with PMCA4b and JAM-A on the Mouse Sperm Flagellum to Regulate Ca2+ Homeostasis and Motility1

PubMed Central

Aravindan, Rolands G.; Fomin, Victor P.; Naik, Ulhas P.; Modelski, Mark J.; Naik, Meghna U.; Galileo, Deni S.; Duncan, Randall L.; Martin-DeLeon, Patricia A.

2012-01-01

Deletion of the highly conserved gene for the major Ca2+ efflux pump, Plasma membrane calcium/calmodulin-dependent ATPase 4b (Pmca4b), in the mouse leads to loss of progressive and hyperactivated sperm motility and infertility. Here we first demonstrate that compared to wild-type (WT), Junctional adhesion molecule-A (Jam-A) null sperm, previously shown to have motility defects and an abnormal mitochondrial phenotype reminiscent of that seen in Pmca4b nulls, exhibit reduced (P<0.001) ATP levels, significantly (P<0.001) greater cytosolic Ca2+ concentration ([Ca2+]c) and ~10-fold higher mitochondrial sequestration, indicating Ca2+ overload. Investigating the mechanism involved, we used coimmunoprecipitation studies to show that CASK (Ca2+/calmodulin-dependent serine kinase), identified for the first time on the sperm flagellum where it co-localizes with both PMCA4b and JAM-A on the proximal principal piece, acts as a common interacting partner of both. Importantly, CASK binds alternatively and non-synergistically with each of these molecules via its single PDZ (PDS-95/Dlg/ZO-1) domain to either inhibit or promote efflux. In the absence of CASK-JAM-A interaction in Jam-A null sperm, CASK-PMCA4b interaction is increased, resulting in inhibition of PMCA4b’s enzymatic activity, consequent Ca2+ accumulation, and a ~6-fold over-expression of constitutively ATP-utilizing CASK, compared to WT. Thus, CASK negatively regulates PMCA4b by directly binding to it and JAM-A positively regulates it indirectly through CASK. The decreased motility is likely due to the collateral net deficit in ATP observed in nulls. Our data indicate that Ca2+ homeostasis in sperm is maintained by the relative ratios of CASK-PMCA4b and CASK-JAM-A interactions. PMID:22020416

Survivability Tests on a Nuclear Waste Cask in Simulated Railroad Accident Fires.

DTIC Science & Technology

1983-06-01

Axial Reference Point ( XRP ) .......... 19 4. A View of the Torch Facility with the Nozzle Directed Side-On to the HNPF Cask... XRP and the TIC for Various HNPF Cask Surfaces in Test Number 1 .................... 47 16. The Spatial Distribution of Sensors in a Cross-Sectional...Plane Through the HNPF Cask at 289.6 cm from the XRP as Viewed from the Top End with the TIC Located at 900 for Test Numbers 1 and 2

Estimation of Inherent Safety Margins in Loaded Commercial Spent Nuclear Fuel Casks

DOE PAGES

Banerjee, Kaushik; Robb, Kevin R.; Radulescu, Georgeta; ...

2016-06-15

We completed a novel assessment to determine the unquantified and uncredited safety margins (i.e., the difference between the licensing basis and as-loaded calculations) available in as-loaded spent nuclear fuel (SNF) casks. This assessment was performed as part of a broader effort to assess issues and uncertainties related to the continued safety of casks during extended storage and transportability following extended storage periods. Detailed analyses crediting the actual as-loaded cask inventory were performed for each of the casks at three decommissioned pressurized water reactor (PWR) sites to determine their characteristics relative to regulatory safety criteria for criticality, thermal, and shielding performance.more » These detailed analyses were performed in an automated fashion by employing a comprehensive and integrated data and analysis tool—Used Nuclear Fuel-Storage, Transportation & Disposal Analysis Resource and Data System (UNF-ST&DARDS). Calculated uncredited criticality margins from 0.07 to almost 0.30 Δk eff were observed; calculated decay heat margins ranged from 4 to almost 22 kW (as of 2014); and significant uncredited transportation dose rate margins were also observed. The results demonstrate that, at least for the casks analyzed here, significant uncredited safety margins are available that could potentially be used to compensate for SNF assembly and canister structural performance related uncertainties associated with long-term storage and subsequent transportation. The results also suggest that these inherent margins associated with how casks are loaded could support future changes in cask licensing to directly or indirectly credit the margins. Work continues to quantify the uncredited safety margins in the SNF casks loaded at other nuclear reactor sites.« less

NRC approves spent-fuel cask for general use: Who needs Yucca Mountain?

DOE Office of Scientific and Technical Information (OSTI.GOV)

Simpson, J.

1993-07-01

The Nuclear Regulatory Commission (NRC) on April 7, 1993, added Pacific Sierra Nuclear Associates`s (PSNA`s) VSC-24 spent-fuel container to its list of approved storage casks. Unlike previously approved designs, however, the cask was made available for use by utilities without site-specific approval. The VSC-24 (ventilated storage cask) is a 130-ton, 16-foot high vertical storage container composed of a ventilated concrete cask (VCC) housing a steel multi-assembly sealed basket (MSB). A third component, a transfer cask (MTC), shields, supports, and protects the MSB during fuel loading and VCC loading operations. The VCC is a cylindrical reinforced-concrete cask 29 inches thick, withmore » a 1.75-inch-thick A 36 steel liner. The cask contains eight vents-four on the top and four on the bottom-to provide for MSB (and fuel rod) cooling. Its concrete shell provides protection against shearing and penetration by tornado projectiles, protects the MSB in the event of a drop or tipover, and is designed to withstand internal temperatures of 350 degrees Farenheit. The VCC is closed with a bolted-down cover of 0.75-inch-thick A 36 steel. The MSB, which provides the primary boundary for 24 spent fuel rods, is a cylindrical steel shell with a thick shield plug and steel cover plates welded at each end. The shell and covers are constructed from SA 516 Grade 70 pressure vessel steel. Fuel is housed in a basket fabricated from SA 516 Grade 70 sheet steel. Penetrations in the MSB`s structural and shield lids allow for vacuum drying and backfilling with helium after fuel loading. Although its manufacturer claims a design life of 50 years, the NRC has licensed the VSC-24 cask for 20 years.« less

Cosmic Ray Muon Imaging of Spent Nuclear Fuel in Dry Storage Casks

DOE PAGES

Durham, J. Matthew; Guardincerri, Elena; Morris, Christopher L.; ...

2016-04-29

In this paper, cosmic ray muon radiography has been used to identify the absence of spent nuclear fuel bundles inside a sealed dry storage cask. The large amounts of shielding that dry storage casks use to contain radiation from the highly radioactive contents impedes typical imaging methods, but the penetrating nature of cosmic ray muons allows them to be used as an effective radiographic probe. This technique was able to successfully identify missing fuel bundles inside a sealed Westinghouse MC-10 cask. This method of fuel cask verification may prove useful for international nuclear safeguards inspectors. Finally, muon radiography may findmore » other safety and security or safeguards applications, such as arms control verification.« less

Verification of Spent Nuclear Fuel in Sealed Dry Storage Casks via Measurements of Cosmic-Ray Muon Scattering

NASA Astrophysics Data System (ADS)

Durham, J. M.; Poulson, D.; Bacon, J.; Chichester, D. L.; Guardincerri, E.; Morris, C. L.; Plaud-Ramos, K.; Schwendiman, W.; Tolman, J. D.; Winston, P.

2018-04-01

Most of the plutonium in the world resides inside spent nuclear reactor fuel rods. This high-level radioactive waste is commonly held in long-term storage within large, heavily shielded casks. Currently, international nuclear safeguards inspectors have no stand-alone method of verifying the amount of reactor fuel stored within a sealed cask. Here we demonstrate experimentally that measurements of the scattering angles of cosmic-ray muons, which pass through a storage cask, can be used to determine if spent fuel assemblies are missing without opening the cask. This application of technology and methods commonly used in high-energy particle physics provides a potential solution to this long-standing problem in international nuclear safeguards.

Sensitivity analysis for best-estimate thermal models of vertical dry cask storage systems

DOE Office of Scientific and Technical Information (OSTI.GOV)

DeVoe, Remy R.; Robb, Kevin R.; Skutnik, Steven E.

Loading requirements for dry cask storage of spent nuclear fuel are driven primarily by decay heat capacity limitations, which themselves are determined through recommended limits on peak cladding temperature within the cask. This study examines the relative sensitivity of peak material temperatures within the cask to parameters that influence both the stored fuel residual decay heat as well as heat removal mechanisms. Here, these parameters include the detailed reactor operating history parameters (e.g., soluble boron concentrations and the presence of burnable poisons) as well as factors that influence heat removal, including non-dominant processes (such as conduction from the fuel basketmore » to the canister and radiation within the canister) and ambient environmental conditions. By examining the factors that drive heat removal from the cask alongside well-understood factors that drive decay heat, it is therefore possible to make a contextual analysis of the most important parameters to evaluation of peak material temperatures within the cask.« less

Cosmic ray muon computed tomography of spent nuclear fuel in dry storage casks

NASA Astrophysics Data System (ADS)

Poulson, D.; Durham, J. M.; Guardincerri, E.; Morris, C. L.; Bacon, J. D.; Plaud-Ramos, K.; Morley, D.; Hecht, A. A.

2017-01-01

Radiography with cosmic ray muon scattering has proven to be a successful method of imaging nuclear material through heavy shielding. Of particular interest is monitoring dry storage casks for diversion of plutonium contained in spent reactor fuel. Using muon tracking detectors that surround a cylindrical cask, cosmic ray muon scattering can be simultaneously measured from all azimuthal angles, giving complete tomographic coverage of the cask interior. This paper describes the first application of filtered back projection algorithms, typically used in medical imaging, to cosmic ray muon scattering imaging. The specific application to monitoring spent nuclear fuel in dry storage casks is investigated via GEANT4 simulations. With a cylindrical muon tracking detector surrounding a typical spent fuel cask, simulations indicate that missing fuel bundles can be detected with a statistical significance of ∼ 18 σ in less than two days exposure and a sensitivity at 1σ to a 5% missing portion of a fuel bundle. Potential detector technologies and geometries are discussed.

Sensitivity analysis for best-estimate thermal models of vertical dry cask storage systems

DOE PAGES

DeVoe, Remy R.; Robb, Kevin R.; Skutnik, Steven E.

2017-07-08

Loading requirements for dry cask storage of spent nuclear fuel are driven primarily by decay heat capacity limitations, which themselves are determined through recommended limits on peak cladding temperature within the cask. This study examines the relative sensitivity of peak material temperatures within the cask to parameters that influence both the stored fuel residual decay heat as well as heat removal mechanisms. Here, these parameters include the detailed reactor operating history parameters (e.g., soluble boron concentrations and the presence of burnable poisons) as well as factors that influence heat removal, including non-dominant processes (such as conduction from the fuel basketmore » to the canister and radiation within the canister) and ambient environmental conditions. By examining the factors that drive heat removal from the cask alongside well-understood factors that drive decay heat, it is therefore possible to make a contextual analysis of the most important parameters to evaluation of peak material temperatures within the cask.« less

Dry transfer system for spent fuel: Project report, A system designed to achieve the dry transfer of bare spent fuel between two casks. Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dawson, D.M.; Guerra, G.; Neider, T.

1995-12-01

This report describes the system developed by EPRI/DOE for the dry transfer of spent fuel assemblies outside the reactor spent fuel pool. The system is designed to allow spent fuel assemblies to be removed from a spent fuel pool in a small cask, transported to the transfer facility, and transferred to a larger cask, either for off-site transportation or on-site storage. With design modifications, this design is capable of transferring single spent fuel assemblies from dry storage casks to transportation casks or visa versa. One incentive for the development of this design is that utilities with limited lifting capacity ormore » other physical or regulatory constraints are limited in their ability to utilize the current, more efficient transportation and storage cask designs. In addition, DOE, in planning to develop and implement the multi-purpose canister (MPC) system for the Civilian Radioactive Waste Management System, included the concept of an on-site dry transfer system to support the implementation of the MPC system at reactors with limitations that preclude the handling of the MPC system transfer casks. This Dry Transfer System can also be used at reactors wi decommissioned spent fuel pools and fuel in dry storage in non-MPC systems to transfer fuel into transportation casks. It can also be used at off-reactor site interim storage facilities for the same purpose.« less

Age Differences in Perseveration: Cognitive and Neuroanatomical Mediators of Performance on the Wisconsin Card Sorting Test

ERIC Educational Resources Information Center

Head, Denise; Kennedy, Kristen M.; Rodrigue, Karen M.; Raz, Naftali

2009-01-01

Aging effects on the Wisconsin Card Sorting Test (WCST) are fairly well established but the mechanisms of the decline are not clearly understood. In this study, we examined the cognitive and neural mechanisms mediating age-related increases in perseveration on the WCST. MRI-based volumetry and measures of selected executive functions in…

78 FR 16619 - List of Approved Spent Fuel Storage Casks: MAGNASTOR® System

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-18

...-0308] RIN 3150-AJ22 List of Approved Spent Fuel Storage Casks: MAGNASTOR[supreg] System AGENCY: Nuclear... proposing to amend its spent fuel storage regulations by revising the NAC International, Inc., Modular Advanced Generation Nuclear All-purpose Storage (MAGNASTOR[supreg]) Cask System listing within the ``List...

Verification of Spent Nuclear Fuel in Sealed Dry Storage Casks via Measurements of Cosmic-Ray Muon Scattering

DOE PAGES

Durham, J. M.; Poulson, D.; Bacon, J.; ...

2018-04-10

Most of the plutonium in the world resides inside spent nuclear reactor fuel rods. This high-level radioactive waste is commonly held in long-term storage within large, heavily shielded casks. Currently, international nuclear safeguards inspectors have no stand-alone method of verifying the amount of reactor fuel stored within a sealed cask. In this paper, we demonstrate experimentally that measurements of the scattering angles of cosmic-ray muons, which pass through a storage cask, can be used to determine if spent fuel assemblies are missing without opening the cask. Finally, this application of technology and methods commonly used in high-energy particle physics providesmore » a potential solution to this long-standing problem in international nuclear safeguards.« less

Verification of Spent Nuclear Fuel in Sealed Dry Storage Casks via Measurements of Cosmic-Ray Muon Scattering

DOE Office of Scientific and Technical Information (OSTI.GOV)

Durham, J. M.; Poulson, D.; Bacon, J.

Most of the plutonium in the world resides inside spent nuclear reactor fuel rods. This high-level radioactive waste is commonly held in long-term storage within large, heavily shielded casks. Currently, international nuclear safeguards inspectors have no stand-alone method of verifying the amount of reactor fuel stored within a sealed cask. In this paper, we demonstrate experimentally that measurements of the scattering angles of cosmic-ray muons, which pass through a storage cask, can be used to determine if spent fuel assemblies are missing without opening the cask. Finally, this application of technology and methods commonly used in high-energy particle physics providesmore » a potential solution to this long-standing problem in international nuclear safeguards.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

J. Bisset

This calculation documents the design of the Spent Nuclear Fuel (SNF) and High-Level Waste (HLW) Cask storage slab for the Aging Area. The design is based on the weights of casks that may be stored on the slab, the weights of vehicles that may be used to move the casks, and the layout shown on the sketch for a 1000 Metric Ton of Heavy Metal (MTHM) storage pad on Attachment 2, Sht.1 of the calculation 170-C0C-C000-00100-000-00A (BSC 2004a). The analytical model used herein is based on the storage area for 8 vertical casks. To simplify the model, the storage areamore » of the horizontal concrete modules and their related shield walls is not included. The heavy weights of the vertical storage casks and the tensile forces due to pullout at the anchorages will produce design moments and shear forces that will envelope those that would occur in the storage area of the horizontal modules. The design loadings will also include snow and live loads. In addition, the design will also reflect pertinent geotechnical data. This calculation will document the preliminary thickness and general reinforcing steel requirements for the slab. This calculation also documents the initial design of the cask anchorage. Other slab details are not developed in this calculation. They will be developed during the final design process. The calculation also does not include the evaluation of the effects of cask drop loads. These will be evaluated in this or another calculation when the exact cask geometry is known.« less

Canister Design for Deep Borehole Disposal of Nuclear Waste

DTIC Science & Technology

2006-05-01

radioactive waste disposal (not yet released) Fortunately, transportation casks for spent fuel have already been approved, built, and used as...would allow use of the current designs for transportation casks ; or, place the fuel assemblies into the final disposal canisters 21 prior to transport ...16 Figure 1-5. Typical Spent Fuel Transportation Casks

Parkin mediates the ubiquitination of VPS35 and modulates retromer-dependent endosomal sorting.

PubMed

Williams, Erin T; Glauser, Liliane; Tsika, Elpida; Jiang, Haisong; Islam, Shariful; Moore, Darren J

2018-06-11

Mutations in a number of genes cause familial forms of Parkinson's disease (PD), including mutations in the vacuolar protein sorting 35 ortholog (VPS35) and parkin genes. In this study, we identify a novel functional interaction between parkin and VPS35. We demonstrate that parkin interacts with and robustly ubiquitinates VPS35 in human neural cells. Familial parkin mutations are impaired in their ability to ubiquitinate VPS35. Parkin mediates the attachment of an atypical poly-ubiquitin chain to VPS35 with three lysine residues identified within the C-terminal region of VPS35 that are covalently modified by ubiquitin. Notably, parkin-mediated VPS35 ubiquitination does not promote the proteasomal degradation of VPS35. Furthermore, parkin does not influence the steady-state levels or turnover of VPS35 in neural cells and VPS35 levels are normal in the brains of parkin knockout mice. These data suggest that ubiquitination of VPS35 by parkin may instead serve a non-degradative cellular function potentially by regulating retromer-dependent sorting. Accordingly, we find that components of the retromer-associated WASH complex are markedly decreased in the brain of parkin knockout mice, suggesting that parkin may modulate WASH complex-dependent retromer sorting. Parkin gene silencing in primary cortical neurons selectively disrupts the vesicular sorting of the autophagy receptor ATG9A, a WASH-dependent retromer cargo. Parkin is not required for dopaminergic neurodegeneration induced by the expression of PD-linked D620N VPS35 in mice, consistent with VPS35 being located downstream of parkin function. Our data reveal a novel functional interaction of parkin with VPS35 that may be important for retromer-mediated endosomal sorting and PD.

[Lentivirus-mediated RNA interference of CD133 inhibits the proliferation of CD133(+) liver cancer stem cells and increases their cisplatin chemosensitivity].

PubMed

Lan, Xi; Wang, Yong; Cao, Shu; Zou, Dongling; Li, Fang; Li, Shaolin

2012-12-01

To study the effects of CD133 suppression by lentivirus-mediated RNA interference (RNAi) on the proliferation and chemosensitivity of CD133(+) cancer stem cells (CSCs) sorted from HepG2 cell line. CD133(+) and CD133- cells were sorted from HepG2 cell line by flow cytometry, and the expression of CD133 before and after cell sorting were detected. The stem cell property of sorted CD133(+) cells were validated by sphere-forming assay in vitro and xenograft experiments in vivo. Lentivirus-mediated short hairpin RNA (shRNA) targeting CD133 were transfected into CD133(+) cells, and CD133 mRNA and protein expressions of the transfected cells were detected by RT-PCR and Western blotting, respectively. Before and after the transfection, the proliferative ability of CD133(+) cells was evaluated by colony formation assay, and the cell growth inhibition rate and apoptosis following cisplatin exposure were detected using CCK-8 assay and flow cytometry. The sorted CD133(+) cells showed a high purity of (88.74∓3.19)%, as compared with the purity of (3.36∓1.80)% before cell sorting. CD133(+) cells showed a high tumor sphere formation ability and tumorigenesis capacity compared with CD133- cells. CD133 shRNA transfection significantly inhibited CD133 mRNA and protein expressions in CD133(+) cells (P<0.01), resulting also in a significantly lowered cell proliferative ability (P<0.01) and an increased growth inhibition rate (P<0.01) and obviously increased cell apoptosis (P<0.05) after cisplatin exposure. Lentivirus-mediated RNAi for CD133 suppression inhibits the proliferation of CD133(+) liver cancer stem cells and increases their chemosensitivity to cisplatin.

Test Plan for the Boiling Water Reactor Dry Cask Simulator

DOE Office of Scientific and Technical Information (OSTI.GOV)

Durbin, Samuel; Lindgren, Eric R.

The thermal performance of commercial nuclear spent fuel dry storage casks are evaluated through detailed numerical analysis . These modeling efforts are completed by the vendor to demonstrate performance and regulatory compliance. The calculations are then independently verified by the Nuclear Regulatory Commission (NRC). Carefully measured data sets generated from testing of full sized casks or smaller cask analogs are widely recognized as vital for validating these models. Recent advances in dry storage cask designs have significantly increased the maximum thermal load allowed in a cask in part by increasing the efficiency of internal conduction pathways and by increasing themore » internal convection through greater canister helium pressure. These same vertical, canistered cask systems rely on ventilation between the canister and the overpack to convect heat away from the canister to the environment for both above and below-ground configurations. While several testing programs have been previously conducted, these earlier validation attempts did not capture the effects of elevated helium pressures or accurately portray the external convection of above-ground and below-ground canistered dry cask systems. The purpose of the investigation described in this report is to produce a data set that can be used to test the validity of the assumptions associated with the calculations presently used to determine steady-state cladding temperatures in modern vertical, canistered dry cask systems. The BWR cask simulator (BCS) has been designed in detail for both the above-ground and below-ground venting configurations. The pressure vessel representing the canister has been designed, fabricated, and pressure tested for a maximum allowable pressure (MAWP) rating of 24 bar at 400 deg C. An existing electrically heated but otherwise prototypic BWR Incoloy-clad test assembly is being deployed inside of a representative storage basket and cylindrical pressure vessel that represents the canister. The symmetric single assembly geometry with well-controlled boundary conditions simplifies interpretation of results. Various configurations of outer concentric ducting will be used to mimic conditions for above and below-ground storage configurations of vertical, dry cask systems with canisters. Radial and axial temperature profiles will be measured for a wide range of decay power and helium cask pressures. Of particular interest is the evaluation of the effect of increased helium pressure on allowable heat load and the effect of simulated wind on a simplified below ground vent configuration. While incorporating the best available information, this test plan is subject to changes due to improved understanding from modeling or from as-built deviations to designs. As-built conditions and actual procedures will be documented in the final test report.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Luna, R. E.

This paper provides a simple model for estimating the release of respirable aerosols resulting from an attack on a spent fuel cask using a high energy density device (HEDD). Two primary experiments have provided data on potential releases from spent fuel casks under HEDD attack. Sandia National Laboratories (SNL) conducted the first in the early 1980's and the second was sponsored by Gessellshaft fur Anlagen- and Reaktorsicherheit (GRS) in Germany and conducted in France in 1994. Both used surrogate spent fuel assemblies in real casks. The SNL experiments used un-pressurized fuel pin assemblies in a single element cask while themore » GRS tests used pressurized fuel pin assemblies in a 9-element cask. Data from the two test programs is reasonably consistent, given the differences in the experiments, but the use of the test data for prediction of releases resulting from HEDD attack requires a method for accounting for the effects of pin pressurization release and the ratio of pin plenum gas release to cask free volume (VR). To account for the effects of VR and to link the two data sources, a simple model has been developed that uses both the SNL data and the GRS data as well as recent test data on aerosols produced in experiments with single pellets subjected to HEDD effects conducted under the aegis of the International Consortium's Working Group on Sabotage of Transport and Storage Casks (WGSTSC). (authors)« less

Monte Carlo Shielding Comparative Analysis Applied to TRIGA HEU and LEU Spent Fuel Transport

NASA Astrophysics Data System (ADS)

Margeanu, C. A.; Margeanu, S.; Barbos, D.; Iorgulis, C.

2010-12-01

The paper is a comparative study of LEU and HEU fuel utilization effects for the shielding analysis during spent fuel transport. A comparison against the measured data for HEU spent fuel, available from the last stage of spent fuel repatriation fulfilled in the summer of 2008, is also presented. All geometrical and material data for the shipping cask were considered according to NAC-LWT Cask approved model. The shielding analysis estimates radiation doses to shipping cask wall surface, and in air at 1 m and 2 m, respectively, from the cask, by means of 3D Monte Carlo MORSE-SGC code. Before loading into the shipping cask, TRIGA spent fuel source terms and spent fuel parameters have been obtained by means of ORIGEN-S code. Both codes are included in ORNL's SCALE 5 programs package. The actinides contribution to total fuel radioactivity is very low in HEU spent fuel case, becoming 10 times greater in LEU spent fuel case. Dose rates for both HEU and LEU fuel contents are below regulatory limits, LEU spent fuel photon dose rates being greater than HEU ones. Comparison between HEU spent fuel theoretical and measured dose rates in selected measuring points shows a good agreement, calculated values being greater than the measured ones both to cask wall surface (about 34% relative difference) and in air at 1 m distance from cask surface (about 15% relative difference).

77 FR 9515 - List of Approved Spent Fuel Storage Casks: HI-STORM 100, Revision 8

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-17

... Storage Casks: HI-STORM 100, Revision 8 AGENCY: Nuclear Regulatory Commission. ACTION: Direct final rule... regulations by revising the Holtec International HI-STORM 100 dry cask storage system listing within the... and safety will be adequately protected. This direct final rule revises the HI-STORM 100 listing in 10...

75 FR 27401 - List of Approved Spent Fuel Storage Casks: NUHOMS® HD System Revision 1; Correction

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-17

... Storage Casks: NUHOMS[reg] HD System Revision 1; Correction AGENCY: Nuclear Regulatory Commission. ACTION... HD spent fuel storage cask system. This action is necessary to correctly specify the effective date... on May 6, 2010 (75 FR 24786), that amends the regulations that govern storage of spent nuclear fuel...

10 CFR 72.240 - Conditions for spent fuel storage cask reapproval.

Code of Federal Regulations, 2011 CFR

2011-01-01

... has been determined by the NRC. The application must be accompanied by a safety analysis report (SAR). The new SAR may reference the SAR originally submitted for the approved spent fuel storage cask design. (c) The design of a spent fuel storage cask will be reapproved if the conditions in § 72.238 are met...

Results for the Aboveground Configuration of the Boiling Water Reactor Dry Cask Simulator

DOE Office of Scientific and Technical Information (OSTI.GOV)

Durbin, Samuel G.; Lindgren, Eric Richard

The thermal performance of commercial nuclear spent fuel dry storage casks are evaluated through detailed numerical analysis. These modeling efforts are completed by the vendor to demonstrate performance and regulatory compliance. The calculations are then independently verified by the Nuclear Regulatory Commission (NRC). Carefully measured data sets generated from testing of full sized casks or smaller cask analogs are widely recognized as vital for validating these models. Recent advances in dry storage cask designs have significantly increased the maximum thermal load allowed in a cask in part by increasing the efficiency of internal conduction pathways and also by increasing themore » internal convection through greater canister helium pressure. These same canistered cask systems rely on ventilation between the canister and the overpack to convect heat away from the canister to the environment for both above and belowground configurations. While several testing programs have been previously conducted, these earlier validation attempts did not capture the effects of elevated helium pressures or accurately portray the external convection of aboveground and belowground canistered dry cask systems. The purpose of the current investigation was to produce data sets that can be used to test the validity of the assumptions associated with the calculations used to determine steady-state cladding temperatures in modern dry casks that utilize elevated helium pressure in the sealed canister in an aboveground configuration. An existing electrically heated but otherwise prototypic BWR Incoloy-clad test assembly was deployed inside of a representative storage basket and cylindrical pressure vessel that represents a vertical canister system. The symmetric single assembly geometry with well-controlled boundary conditions simplifies interpretation of results. The arrangement of ducting was used to mimic conditions for an aboveground storage configuration in a vertical, dry cask systems with canisters. Transverse and axial temperature profiles were measured for a wide range of decay power and helium cask pressures. Of particular interest was the evaluation of the effect of increased helium pressure on peak cladding temperatures (PCTs) for identical thermal loads. All steady state peak temperatures and induced flow rates increased with increasing assembly power. Peak cladding temperatures decreased with increasing internal helium pressure for a given assembly power, indicating increased internal convection. In addition, the location of the PCT moved from near the top of the assembly to ~1/3 the height of the assembly for the highest (8 bar absolute) to the lowest (0 bar absolute) pressure studied, respectively. This shift in PCT location is consistent with the varying contribution of convective heat transfer proportional with of internal helium pressure.« less

Tandem SAM Domain Structure of Human Caskin1: A Presynaptic, Self-Assembling Scaffold for CASK

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stafford, Ryan L.; Hinde, Elizabeth; Knight, Mary Jane

2012-02-07

The synaptic scaffolding proteins CASK and Caskin1 are part of the fibrous mesh of proteins that organize the active zones of neural synapses. CASK binds to a region of Caskin1 called the CASK interaction domain (CID). Adjacent to the CID, Caskin1 contains two tandem sterile a motif (SAM) domains. Many SAM domains form polymers so they are good candidates for forming the fibrous structures seen in the active zone. We show here that the SAM domains of Caskin1 form a new type of SAM helical polymer. The Caskin1 polymer interface exhibits a remarkable segregation of charged residues, resulting in amore » high sensitivity to ionic strength in vitro. The Caskin1 polymers can be decorated with CASK proteins, illustrating how these proteins may work together to organize the cytomatrix in active zones.« less

Evaluation of Cask Drop Criticality Issues at K Basin

DOE Office of Scientific and Technical Information (OSTI.GOV)

GOLDMANN, L.H.

An analysis of ability of Multi-canister Overpack (MCO) to withstand drops at K Basin without exceeding the criticality design requirements. Report concludes the MCO will function acceptably. The spent fuel currently residing in the 105 KE and 105 KW storage basins will be placed in fuel storage baskets which will be loaded into the MCO cask assembly. During the basket loading operations the MCO cask assembly will be positioned near the bottom of the south load out pit (SLOP). The loaded MCO cask will be lifted from the SLOP transferred to the transport trailer and delivered to the Cold Vacuummore » Drying Facility (CVDF). In the wet condition there is a potential for criticality problems if significant changes in the designed fuel configurations occur. The purpose of this report is to address structural issues associated with criticality design features for MCO cask drop accidents in the 105 KE and 105 KW facilities.« less

76 FR 17037 - List of Approved Spent Fuel Storage Casks: HI-STORM Flood/Wind Addition

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-28

...-0007] RIN 3150-AI90 List of Approved Spent Fuel Storage Casks: HI-STORM Flood/Wind Addition AGENCY... or the Commission) is proposing to amend its spent fuel storage cask regulations to add the HI-STORM...: June 13, 2011. SAR Submitted by: Holtec International, Inc. SAR Title: Safety Analysis Report on the HI...

AUTOCASK (AUTOmatic Generation of 3-D CASK models). A microcomputer based system for shipping cask design review analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gerhard, M.A.; Sommer, S.C.

1995-04-01

AUTOCASK (AUTOmatic Generation of 3-D CASK models) is a microcomputer-based system of computer programs and databases developed at the Lawrence Livermore National Laboratory (LLNL) for the structural analysis of shipping casks for radioactive material. Model specification is performed on the microcomputer, and the analyses are performed on an engineering workstation or mainframe computer. AUTOCASK is based on 80386/80486 compatible microcomputers. The system is composed of a series of menus, input programs, display programs, a mesh generation program, and archive programs. All data is entered through fill-in-the-blank input screens that contain descriptive data requests.

Mutated CaV2.1 channels dysregulate CASK/P2X3 signaling in mouse trigeminal sensory neurons of R192Q Cacna1a knock-in mice.

PubMed

Gnanasekaran, Aswini; Bele, Tanja; Hullugundi, Swathi; Simonetti, Manuela; Ferrari, Michael D; van den Maagdenberg, Arn M J M; Nistri, Andrea; Fabbretti, Elsa

2013-12-02

ATP-gated P2X3 receptors of sensory ganglion neurons are important transducers of pain as they adapt their expression and function in response to acute and chronic nociceptive signals. The present study investigated the role of calcium/calmodulin-dependent serine protein kinase (CASK) in controlling P2X3 receptor expression and function in trigeminal ganglia from Cacna1a R192Q-mutated knock-in (KI) mice, a genetic model for familial hemiplegic migraine type-1. KI ganglion neurons showed more abundant CASK/P2X3 receptor complex at membrane level, a result that likely originated from gain-of-function effects of R192Q-mutated CaV2.1 channels and downstream enhanced CaMKII activity. The selective CaV2.1 channel blocker ω-Agatoxin IVA and the CaMKII inhibitor KN-93 were sufficient to return CASK/P2X3 co-expression to WT levels. After CASK silencing, P2X3 receptor expression was decreased in both WT and KI ganglia, supporting the role of CASK in P2X3 receptor stabilization. This process was functionally observed as reduced P2X3 receptor currents. We propose that, in trigeminal sensory neurons, the CASK/P2X3 complex has a dynamic nature depending on intracellular calcium and related signaling, that are enhanced in a transgenic mouse model of genetic hemiplegic migraine.

78 FR 63375 - List of Approved Spent Fuel Storage Casks: Transnuclear, Inc. Standardized NUHOMS® Cask System

Federal Register 2010, 2011, 2012, 2013, 2014

2013-10-24

... include: adding a new transfer cask (TC), the OS197L, for use with the 32PT and 61BT dry shielded.... 1004. Specifically, Transnuclear, Inc. requested changes to: (1) add a new TC, the OS197L, for use with... with NUREG-1745 requirements. Deleting the TC dose rates for all currently licensed payloads (TSs 1.2...

Cosmic ray muons for spent nuclear fuel monitoring

NASA Astrophysics Data System (ADS)

Chatzidakis, Stylianos

There is a steady increase in the volume of spent nuclear fuel stored on-site (at reactor) as currently there is no permanent disposal option. No alternative disposal path is available and storage of spent nuclear fuel in dry storage containers is anticipated for the near future. In this dissertation, a capability to monitor spent nuclear fuel stored within dry casks using cosmic ray muons is developed. The motivation stems from the need to investigate whether the stored content agrees with facility declarations to allow proliferation detection and international treaty verification. Cosmic ray muons are charged particles generated naturally in the atmosphere from high energy cosmic rays. Using muons for proliferation detection and international treaty verification of spent nuclear fuel is a novel approach to nuclear security that presents significant advantages. Among others, muons have the ability to penetrate high density materials, are freely available, no radiological sources are required and consequently there is a total absence of any artificial radiological dose. A methodology is developed to demonstrate the applicability of muons for nuclear nonproliferation monitoring of spent nuclear fuel dry casks. Purpose is to use muons to differentiate between spent nuclear fuel dry casks with different amount of loading, not feasible with any other technique. Muon scattering and transmission are used to perform monitoring and imaging of the stored contents of dry casks loaded with spent nuclear fuel. It is shown that one missing fuel assembly can be distinguished from a fully loaded cask with a small overlapping between the scattering distributions with 300,000 muons or more. A Bayesian monitoring algorithm was derived to allow differentiation of a fully loaded dry cask from one with a fuel assembly missing in the order of minutes and negligible error rate. Muon scattering and transmission simulations are used to reconstruct the stored contents of sealed dry casks from muon measurements. A combination of muon scattering and muon transmission imaging can improve resolution and thus a missing fuel assembly can be identified for vertical and horizontal dry casks. The apparent separation of the images reveals that the muon scattering and transmission can be used for discrimination between casks, satisfying the diversion criteria set by IAEA.

Cosmic ray muon computed tomography of spent nuclear fuel in dry storage casks

DOE PAGES

Poulson, Daniel Cris; Durham, J. Matthew; Guardincerri, Elena; ...

2016-10-22

Radiography with cosmic ray muon scattering has proven to be a successful method of imaging nuclear material through heavy shielding. Of particular interest is monitoring dry storage casks for diversion of plutonium contained in spent reactor fuel. Using muon tracking detectors that surround a cylindrical cask, cosmic ray muon scattering can be simultaneously measured from all azimuthal angles, giving complete tomographic coverage of the cask interior. This article describes the first application of filtered back projection algorithms, typically used in medical imaging, to cosmic ray muon scattering imaging. The specific application to monitoring spent nuclear fuel in dry storage casksmore » is investigated via GEANT4 simulations. With a cylindrical muon tracking detector surrounding a typical spent fuel cask, simulations indicate that missing fuel bundles can be detected with a statistical significance of ~18σ in less than two days exposure and a sensitivity at 1σ to a 5% missing portion of a fuel bundle. Finally, we discuss potential detector technologies and geometries.« less

NEUTRON CHARACTERIZATION OF ENSA-DPT TYPE SPENT FUEL CASK AT TRILLO NUCLEAR POWER PLANT.

PubMed

Méndez-Villafañe, Roberto; Campo-Blanco, Xandra; Embid, Miguel; Yéboles, César A; Morales, Ramón; Novo, Manuel; Sanz, Javier

2018-04-23

The Neutron Standards Laboratory of CIEMAT has conducted the characterization of the independent spent fuel storage installation at the Trillo Nuclear Power Plant. At this facility, the spent fuel assemblies are stored in ENSA-DPT type dual purpose casks. Neutron characterization was performed by dosimetry measurements with a neutron survey meter (LB6411) inside the facility, around an individual cask and between stored casks, and outside the facility. Spectra measurements were also performed with a Bonner sphere system in order to determine the integral quantities and validate the use of the neutron monitor at the different positions. Inside the facility, measured neutron spectra and neutron ambient dose equivalent rate are consistent with the casks spatial distribution and neutron emission rates, and measurements with both instruments are consistent with each other. Outside the facility, measured neutron ambient dose equivalent rates are well below the 0.5 μSv/h limit established by the nuclear regulatory authority.

Cosmic ray muon computed tomography of spent nuclear fuel in dry storage casks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Poulson, Daniel Cris; Durham, J. Matthew; Guardincerri, Elena

Radiography with cosmic ray muon scattering has proven to be a successful method of imaging nuclear material through heavy shielding. Of particular interest is monitoring dry storage casks for diversion of plutonium contained in spent reactor fuel. Using muon tracking detectors that surround a cylindrical cask, cosmic ray muon scattering can be simultaneously measured from all azimuthal angles, giving complete tomographic coverage of the cask interior. This article describes the first application of filtered back projection algorithms, typically used in medical imaging, to cosmic ray muon scattering imaging. The specific application to monitoring spent nuclear fuel in dry storage casksmore » is investigated via GEANT4 simulations. With a cylindrical muon tracking detector surrounding a typical spent fuel cask, simulations indicate that missing fuel bundles can be detected with a statistical significance of ~18σ in less than two days exposure and a sensitivity at 1σ to a 5% missing portion of a fuel bundle. Finally, we discuss potential detector technologies and geometries.« less

SCANS (Shipping Cask ANalysis System) a microcomputer-based analysis system for shipping cask design review: User`s manual to Version 3a. Volume 1, Revision 2

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mok, G.C.; Thomas, G.R.; Gerhard, M.A.

SCANS (Shipping Cask ANalysis System) is a microcomputer-based system of computer programs and databases developed at the Lawrence Livermore National Laboratory (LLNL) for evaluating safety analysis reports on spent fuel shipping casks. SCANS is an easy-to-use system that calculates the global response to impact loads, pressure loads and thermal conditions, providing reviewers with an independent check on analyses submitted by licensees. SCANS is based on microcomputers compatible with the IBM-PC family of computers. The system is composed of a series of menus, input programs, cask analysis programs, and output display programs. All data is entered through fill-in-the-blank input screens thatmore » contain descriptive data requests. Analysis options are based on regulatory cases described in the Code of Federal Regulations 10 CFR 71 and Regulatory Guides published by the US Nuclear Regulatory Commission in 1977 and 1978.« less

Characterization of neutron sources from spent fuel casks. [Skyshine

DOE Office of Scientific and Technical Information (OSTI.GOV)

Parks, C.V.; Pace, J.V. III

1987-01-01

In the interim period prior to the acceptance of spent fuel for disposal by the USDOE, utilities are beginning to choose dry cask storage as an alternative to pool re-racking, transshipments, or new pool construction. In addition, the current MRS proposal calls for interim dry storage of consolidated spent fuel in concrete casks. As part of the licensing requirements for these cask storage facilities, calculations are typically necessary to determine the yearly radiation dose received at the site boundary. Unlike wet facilities, neutron skyshine can be an important contribution to the total boundary dose from a dry storage facility. Calculationmore » of the neutron skyshine is in turn heavily dependent on the source characteristics and source model selected for the analysis. This paper presents the basic source characteristics of the spent fuel stored in dry casks and discusses factors that must be considered in evaluating and modeling the radiation sources for the subsequent skyshine calculation. 4 refs., 1 tab.« less

High energy neutron transmission analysis of dry cask storage

NASA Astrophysics Data System (ADS)

Greulich, Christopher; Hughes, Christopher; Gao, Yuan; Enqvist, Andreas; Baciak, James

2017-12-01

Since the U.S. currently only approves of storing used nuclear fuel in pools or dry casks, the demand for dry cask storage is on the rise due to the continuous operation of currently existing nuclear plants which are reaching or have reached the capacity of their used fuel pools. With the rising demand comes additional pressure to ensure the integrity of dry cask systems. Visual inspection is costly and man-power intensive, so alternative nondestructive testing techniques are desired to insure the continued safe and effective storage of fuel. One such approach being investigated by the University of Florida is neutron based computed tomography. Simulations in MCNP are preformed where D-T energy neutrons are transmitted through the dry cask and measured on the opposite side. If the transmitted signal is clear enough, the interior of the cask can be reconstructed from the measurement of the alterations of neutron signal intensity using standard mathematical techniques developed for medical imaging. Preliminary efforts show a correlation between energy and number of scatters (which is an indication of retention of position information). Work is ongoing to quantify if the correlation is strong enough that an energy discriminator may be used as a filter in future image reconstruction. The calculated transmission probability suggests that an image could be reconstructed with a week of scanning.

Research on Spent Fuel Storage and Transportation in CRIEPI (Part 2 Concrete Cask Storage)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Koji Shirai; Jyunichi Tani; Taku Arai

2008-10-01

Concrete cask storage has been implemented in the world. At a later stage of storage period, the containment of the canister may deteriorate due to stress corrosion cracking phenomena in a salty air environment. High resistant stainless steels against SCC have been tested as compared with normal stainless steel. Taking account of the limited time-length of environment with certain level of humidity and temperature range, the high resistant stainless steels will survive from SCC damage. In addition, the adhesion of salt from salty environment on the canister surface will be further limited with respect to the canister temperature and anglemore » of the canister surface against the salty air flow in the concrete cask. Optional countermeasure against SCC with respect to salty air environment has been studied. Devices consisting of various water trays to trap salty particles from the salty air were designed to be attached at the air inlet for natural cooling of the cask storage building. Efficiency for trapping salty particles was evaluated. Inspection of canister surface was carried out using an optical camera inserted from the air outlet through the annulus of a concrete cask that has stored real spent fuel for more than 15 years. The camera image revealed no gross degradation on the surface of the canister. Seismic response of a full-scale concrete cask with simulated spent fuel assemblies has been demonstrated. The cask did not tip over, but laterally moved by the earthquake motion. Stress generated on the surface of the spent fuel assemblies during the earthquake motion were within the elastic region.« less

Mutated CaV2.1 channels dysregulate CASK/P2X3 signaling in mouse trigeminal sensory neurons of R192Q Cacna1a knock-in mice

PubMed Central

2013-01-01

Background ATP-gated P2X3 receptors of sensory ganglion neurons are important transducers of pain as they adapt their expression and function in response to acute and chronic nociceptive signals. The present study investigated the role of calcium/calmodulin-dependent serine protein kinase (CASK) in controlling P2X3 receptor expression and function in trigeminal ganglia from Cacna1a R192Q-mutated knock-in (KI) mice, a genetic model for familial hemiplegic migraine type-1. Results KI ganglion neurons showed more abundant CASK/P2X3 receptor complex at membrane level, a result that likely originated from gain-of-function effects of R192Q-mutated CaV2.1 channels and downstream enhanced CaMKII activity. The selective CaV2.1 channel blocker ω-Agatoxin IVA and the CaMKII inhibitor KN-93 were sufficient to return CASK/P2X3 co-expression to WT levels. After CASK silencing, P2X3 receptor expression was decreased in both WT and KI ganglia, supporting the role of CASK in P2X3 receptor stabilization. This process was functionally observed as reduced P2X3 receptor currents. Conclusions We propose that, in trigeminal sensory neurons, the CASK/P2X3 complex has a dynamic nature depending on intracellular calcium and related signaling, that are enhanced in a transgenic mouse model of genetic hemiplegic migraine. PMID:24294842

Thermal-Hydraulic Results for the Boiling Water Reactor Dry Cask Simulator.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Durbin, Samuel; Lindgren, Eric R.

The thermal performance of commercial nuclear spent fuel dry storage casks is evaluated through detailed numerical analysis. These modeling efforts are completed by the vendor to demonstrate performance and regulatory compliance. The calculations are then independently verified by the Nuclear Regulatory Commission (NRC). Carefully measured data sets generated from testing of full sized casks or smaller cask analogs are widely recognized as vital for validating these models. Recent advances in dry storage cask designs have significantly increased the maximum thermal load allowed in a cask in part by increasing the efficiency of internal conduction pathways and by increasing the internalmore » convection through greater canister helium pressure. These same canistered cask systems rely on ventilation between the canister and the overpack to convect heat away from the canister to the environment for both aboveground and belowground configurations. While several testing programs have been previously conducted, these earlier validation attempts did not capture the effects of elevated helium pressures or accurately portray the external convection of aboveground and belowground canistered dry cask systems. The purpose of this investigation was to produce validation-quality data that can be used to test the validity of the modeling presently used to determine cladding temperatures in modern vertical dry casks. These cladding temperatures are critical to evaluate cladding integrity throughout the storage cycle. To produce these data sets under well-controlled boundary conditions, the dry cask simulator (DCS) was built to study the thermal-hydraulic response of fuel under a variety of heat loads, internal vessel pressures, and external configurations. An existing electrically heated but otherwise prototypic BWR Incoloy-clad test assembly was deployed inside of a representative storage basket and cylindrical pressure vessel that represents a vertical canister system. The symmetric single assembly geometry with well-controlled boundary conditions simplified interpretation of results. Two different arrangements of ducting were used to mimic conditions for aboveground and belowground storage configurations for vertical, dry cask systems with canisters. Transverse and axial temperature profiles were measured throughout the test assembly. The induced air mass flow rate was measured for both the aboveground and belowground configurations. In addition, the impact of cross-wind conditions on the belowground configuration was quantified. Over 40 unique data sets were collected and analyzed for these efforts. Fourteen data sets for the aboveground configuration were recorded for powers and internal pressures ranging from 0.5 to 5.0 kW and 0.3 to 800 kPa absolute, respectively. Similarly, fourteen data sets were logged for the belowground configuration starting at ambient conditions and concluding with thermal-hydraulic steady state. Over thirteen tests were conducted using a custom-built wind machine. The results documented in this report highlight a small, but representative, subset of the available data from this test series. This addition to the dry cask experimental database signifies a substantial addition of first-of-a-kind, high-fidelity transient and steady-state thermal-hydraulic data sets suitable for CFD model validation.« less

Shielding calculation and criticality safety analysis of spent fuel transportation cask in research reactors.

PubMed

Mohammadi, A; Hassanzadeh, M; Gharib, M

2016-02-01

In this study, shielding calculation and criticality safety analysis were carried out for general material testing reactor (MTR) research reactors interim storage and relevant transportation cask. During these processes, three major terms were considered: source term, shielding, and criticality calculations. The Monte Carlo transport code MCNP5 was used for shielding calculation and criticality safety analysis and ORIGEN2.1 code for source term calculation. According to the results obtained, a cylindrical cask with body, top, and bottom thicknesses of 18, 13, and 13 cm, respectively, was accepted as the dual-purpose cask. Furthermore, it is shown that the total dose rates are below the normal transport criteria that meet the standards specified. Copyright © 2015 Elsevier Ltd. All rights reserved.

TN International and ITS operational feedback regarding the decommissioning of obsolete casks dedicated to the transport and/or storage of nuclear raw materials, fuel and used fuel

DOE Office of Scientific and Technical Information (OSTI.GOV)

Blachet, L.; Bimet, F.; Rennesson, N.

2008-07-01

Within the AREVA group, TN International is a major actor regarding the design of casks and transportation for the nuclear cycle. In the early 2005, TN International has started the project of decommissioning some of its own equipment and was hence the first company ever in the AREVA Group to implement this new approach. In order to do so, TN International has based this project by taking into account the AREVA Sustainable Development Charter, the French regulatory framework, the ANDRA (Agence Nationale pour la Gestion des Dechets Radioactifs - National Agency for the radioactive waste management) requirements and has deployedmore » a step by step methodology such as radiological characterization following a logical route. The aim was to define a standardized process with optimized solutions regarding the diversity of the cask's fleet. As a general matter, decommissioning of nuclear casks is a brand new field as the nuclear field is more familiar with the dismantling of nuclear facilities and/or nuclear power plant. Nevertheless existing workshops, maintenance facilities, measurements equipments and techniques have been exploited and adapted by TN International in order to turn an ambitious project into a permanent and cost-effective activity. The decommissioning of the nuclear casks implemented by TN International regarding its own needs and the French regulatory framework is formalized by several processes and is materialized for instance by the final disposal of casks as they are or in ISO container packed with cut-off casks and big bags filled with crushed internal cask equipments, etc. The first part of this paper aims to describe the history of the project that started with a specific environmental analysis which took into account the values of AREVA as regards the Sustainable Development principles that were at the time and are still a topic of current concern in the world. The second part will deal with the definition, the design and the implementation of the decommissioning processes and the applied techniques. The third part will present a two years operational feedback. The last part will introduce new processes which are currently under investigation and will put into light that decommissioning of nuclear casks is a continuous activity that is in perpetual mutation. (authors)« less

Interaction of cosmic ray muons with spent nuclear fuel dry casks and determination of lower detection limit

NASA Astrophysics Data System (ADS)

Chatzidakis, S.; Choi, C. K.; Tsoukalas, L. H.

2016-08-01

The potential non-proliferation monitoring of spent nuclear fuel sealed in dry casks interacting continuously with the naturally generated cosmic ray muons is investigated. Treatments on the muon RMS scattering angle by Moliere, Rossi-Greisen, Highland and, Lynch-Dahl were analyzed and compared with simplified Monte Carlo simulations. The Lynch-Dahl expression has the lowest error and appears to be appropriate when performing conceptual calculations for high-Z, thick targets such as dry casks. The GEANT4 Monte Carlo code was used to simulate dry casks with various fuel loadings and scattering variance estimates for each case were obtained. The scattering variance estimation was shown to be unbiased and using Chebyshev's inequality, it was found that 106 muons will provide estimates of the scattering variances that are within 1% of the true value at a 99% confidence level. These estimates were used as reference values to calculate scattering distributions and evaluate the asymptotic behavior for small variations on fuel loading. It is shown that the scattering distributions between a fully loaded dry cask and one with a fuel assembly missing initially overlap significantly but their distance eventually increases with increasing number of muons. One missing fuel assembly can be distinguished from a fully loaded cask with a small overlapping between the distributions which is the case of 100,000 muons. This indicates that the removal of a standard fuel assembly can be identified using muons providing that enough muons are collected. A Bayesian algorithm was developed to classify dry casks and provide a decision rule that minimizes the risk of making an incorrect decision. The algorithm performance was evaluated and the lower detection limit was determined.

Risk Assessment of Structural Integrity of Transportation Casks after Extended Storage

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ibarra, Luis; Medina, Ricardo; Yang, Haori

This study assessed the risk of loss of structural integrity of transportation casks and fuel cladding after extended storage. Although it is known that fuel rods discharged from NPPs have a small percentage of rod cladding defects, the behavior of fuel cladding and the structural elements of assemblies during transportation after long-term storage is not well understood. If the fuel degrades during extended storage, it could be susceptible to damage from vibration and impact loads during transport operations, releasing fission-product gases into the canister or the cask interior (NWTRB 2010). Degradation of cladding may occur due to mechanisms associated withmore » hydrogen embrittlement, delayed hydride cracking, low temperature creep, and stress corrosion cracking (SCC) that may affect fuel cladding and canister components after extended storage of hundreds of years. Over extended periods at low temperatures, these mechanisms affect the ductility, strength, and fracture toughness of the fuel cladding, which becomes brittle. For transportation purposes, the fuel may be transferred from storage to shipping casks, or dual-purpose casks may be used for storage and transportation. Currently, most of the transportation casks will be the former case. A risk assessment evaluation is conducted based on results from experimental tests and simulations with advanced numerical models. A novel contribution of this study is the evaluation of the combined effect of component aging and vibration/impact loads in transportation scenarios. The expected levels of deterioration will be obtained from previous and current studies on the effect of aging on fuel and cask components. The emphasis of the study is placed on the structural integrity of fuel cladding and canisters.« less

Binding of Y-P30 to Syndecan 2/3 Regulates the Nuclear Localization of CASK

PubMed Central

Landgraf, Peter; Mikhaylova, Marina; Macharadze, Tamar; Borutzki, Corinna; Zenclussen, Ana-Claudia; Wahle, Petra; Kreutz, Michael R.

2014-01-01

The survival promoting peptide Y-P30 has documented neuroprotective effects as well as cell survival and neurite outgrowth promoting activity in vitro and in vivo. Previous work has shown that multimerization of the peptide with pleiotrophin (PTN) and subsequent binding to syndecan (SDC) -2 and -3 is involved in its neuritogenic effects. In this study we show that Y-P30 application regulates the nuclear localization of the SDC binding partner Calcium/calmodulin-dependent serine kinase (CASK) in neuronal primary cultures during development. In early development at day in vitro (DIV) 8 when mainly SDC-3 is expressed supplementation of the culture medium with Y-P30 reduces nuclear CASK levels whereas it has the opposite effect at DIV 18 when SDC-2 is the dominant isoform. In the nucleus CASK regulates gene expression via its association with the T-box transcription factor T-brain-1 (Tbr-1) and we indeed found that gene expression of downstream targets of this complex, like the GluN2B NMDA-receptor, exhibits a corresponding down- or up-regulation at the mRNA level. The differential effect of Y-P30 on the nuclear localization of CASK correlates with its ability to induce shedding of the ectodomain of SDC-2 but not -3. shRNA knockdown of SDC-2 at DIV 18 and SDC-3 at DIV 8 completely abolished the effect of Y-P30 supplementation on nuclear CASK levels. During early development a protein knockdown of SDC-3 also attenuated the effect of Y-P30 on axon outgrowth. Taken together these data suggest that Y-P30 can control the nuclear localization of CASK in a SDC-dependent manner. PMID:24498267

Inspection and Gamma-Ray Dose Rate Measurements of the Annulus of the VSC-17 Concrete Spent Nuclear Fuel Storage Cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

P. L. Winston

2007-09-01

The air cooling annulus of the Ventilated Storage Cask (VSC)-17 spent fuel storage cask was inspected using a Toshiba 7 mm (1/4”) CCD video camera. The dose rates observed in the annular space were measured to provide a reference for the activity to which the camera(s) being tested were being exposed. No gross degradation, pitting, or general corrosion was observed.

Greater-than-Class C low-level radioactive waste shipping package/container identification and requirements study. National Low-Level Waste Management Program

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tyacke, M.

1993-08-01

This report identifies a variety of shipping packages (also referred to as casks) and waste containers currently available or being developed that could be used for greater-than-Class C (GTCC) low-level waste (LLW). Since GTCC LLW varies greatly in size, shape, and activity levels, the casks and waste containers that could be used range in size from small, to accommodate a single sealed radiation source, to very large-capacity casks/canisters used to transport or dry-store highly radioactive spent fuel. In some cases, the waste containers may serve directly as shipping packages, while in other cases, the containers would need to be placedmore » in a transport cask. For the purpose of this report, it is assumed that the generator is responsible for transporting the waste to a Department of Energy (DOE) storage, treatment, or disposal facility. Unless DOE establishes specific acceptance criteria, the receiving facility would need the capability to accept any of the casks and waste containers identified in this report. In identifying potential casks and waste containers, no consideration was given to their adequacy relative to handling, storage, treatment, and disposal. Those considerations must be addressed separately as the capabilities of the receiving facility and the handling requirements and operations are better understood.« less

GGA3 mediates TrkA endocytic recycling to promote sustained Akt phosphorylation and cell survival

PubMed Central

Li, Xuezhi; Lavigne, Pierre; Lavoie, Christine

2015-01-01

Although TrkA postendocytic sorting significantly influences neuronal cell survival and differentiation, the molecular mechanism underlying TrkA receptor sorting in the recycling or degradation pathways remains poorly understood. Here we demonstrate that Golgi-localized, γ adaptin-ear–containing ADP ribosylation factor-binding protein 3 (GGA3) interacts directly with the TrkA cytoplasmic tail through an internal DXXLL motif and mediates the functional recycling of TrkA to the plasma membrane. We find that GGA3 depletion by siRNA delays TrkA recycling, accelerates TrkA degradation, attenuates sustained NGF-induced Akt activation, and reduces cell survival. We also show that GGA3’s effect on TrkA recycling is dependent on the activation of Arf6. This work identifies GGA3 as a key player in a novel DXXLL-mediated endosomal sorting machinery that targets TrkA to the plasma membrane, where it prolongs the activation of Akt signaling and survival responses. PMID:26446845

Radioactive materials shipping cask anticontamination enclosure

DOEpatents

Belmonte, Mark S.; Davis, James H.; Williams, David A.

1982-01-01

An anticontamination device for use in storing shipping casks for radioactive materials comprising (1) a seal plate assembly; (2) a double-layer plastic bag; and (3) a water management system or means for water management.

Impact of Reactor Operating Parameters on Cask Reactivity in BWR Burnup Credit

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ilas, Germina; Betzler, Benjamin R; Ade, Brian J

This paper discusses the effect of reactor operating parameters used in fuel depletion calculations on spent fuel cask reactivity, with relevance for boiling-water reactor (BWR) burnup credit (BUC) applications. Assessments that used generic BWR fuel assembly and spent fuel cask configurations are presented. The considered operating parameters, which were independently varied in the depletion simulations for the assembly, included fuel temperature, bypass water density, specific power, and operating history. Different operating history scenarios were considered for the assembly depletion to determine the effect of relative power distribution during the irradiation cycles, as well as the downtime between cycles. Depletion, decay,more » and criticality simulations were performed using computer codes and associated nuclear data within the SCALE code system. Results quantifying the dependence of cask reactivity on the assembly depletion parameters are presented herein.« less

Safety Analysis of Dual Purpose Metal Cask Subjected to Impulsive Loads due to Aircraft Engine Crash

NASA Astrophysics Data System (ADS)

Shirai, Koji; Namba, Kosuke; Saegusa, Toshiari

In Japan, the first Interim Storage Facility of spent nuclear fuel away from reactor site is being planned to start its commercial operation around 2010, in use of dual-purpose metal cask in the northern part of Main Japan Island. Business License Examination for safety design approval has started since March, 2007. To demonstrate the more scientific and rational performance of safety regulation activities on each phase for the first license procedure, CREPEI has executed demonstration tests with full scale casks, such as drop tests onto real targets without impact limiters(1) and seismic tests subjected to strong earthquake motions(2). Moreover, it is important to develop the knowledge for the inherent security of metal casks under extreme mechanical-impact conditions, especially for increasing interest since the terrorist attacks from 11th September 2001(3)-(6). This paper presents dynamic mechanical behavior of the metal cask lid closure system caused by direct aircraft engine crash and describes calculated results (especially, leak tightness based on relative dynamic displacements between metallic seals). Firstly, the local penetration damage of the interim storage facility building by a big passenger aircraft engine crash (diameter 2.7m, length 4.3m, weight 4.4ton, impact velocity 90m/s) has been examined. The reduced velocity is calculated by the local damage formula for concrete structure with its thickness of 70cm. The load vs. time function for this reduced velocity (60m/s) is estimated by the impact analysis using Finite Element code LS-DYNA with the full scale engine model onto a hypothetically rigid target. Secondly, as the most critical scenarios for the metal cask, two impact scenarios (horizontal impact hitting the cask and vertical impact onto the lid metallic seal system) are chosen. To consider the geometry of all bolts for two lids, the gasket reaction forces and the inner pressure of the cask cavity, the detailed three dimensional FEM models are developed and calculated. Main criteria for estimating the maximum leakage rate for the lid metallic seal system are no loss of the pre-stress of the lid bolts, no appearance of the plastic region between the metal seal flanges, and no large relative deformation of the lid seals. Finally, in both cases, the low leakage rate for the metal cask lid closure system under the impulsive loads due to aircraft engine crash will be proved thoroughly.

Preliminary design report: Babcock and Wilcox BR-100 100-ton rail/barge spent fuel shipping cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

None

1990-02-01

The purpose of this document is to provide information on burnup credit as applied to the preliminary design of the BR-100 shipping cask. There is a brief description of the preliminary basket design and the features used to maintain a critically safe system. Following the basket description is a discussion of various criticality analyses used to evaluate burnup credit. The results from these analyses are then reviewed in the perspective of fuel burnups expected to be shipped to either the final repository or a Monitored Retrievable Storage (MRS) facility. The hurdles to employing burnup credit in the certification of anymore » cask are then outlines and reviewed. the last section gives conclusions reached as to burnup credit for the BR-100 cask, based on our analyses and experience. All information in this study refers to the cask configured to transport PWR fuel. Boiling Water Reactor (BWR) fuel satisfies the criticality requirements so that burnup credit is not needed. All calculations generated in the preparation of this report were based upon the preliminary design which will be optimized during the final design. 8 refs., 19 figs., 16 tabs.« less

Evaluation of RAPID for a UNF cask benchmark problem

NASA Astrophysics Data System (ADS)

Mascolino, Valerio; Haghighat, Alireza; Roskoff, Nathan J.

2017-09-01

This paper examines the accuracy and performance of the RAPID (Real-time Analysis for Particle transport and In-situ Detection) code system for the simulation of a used nuclear fuel (UNF) cask. RAPID is capable of determining eigenvalue, subcritical multiplication, and pin-wise, axially-dependent fission density throughout a UNF cask. We study the source convergence based on the analysis of the different parameters used in an eigenvalue calculation in the MCNP Monte Carlo code. For this study, we consider a single assembly surrounded by absorbing plates with reflective boundary conditions. Based on the best combination of eigenvalue parameters, a reference MCNP solution for the single assembly is obtained. RAPID results are in excellent agreement with the reference MCNP solutions, while requiring significantly less computation time (i.e., minutes vs. days). A similar set of eigenvalue parameters is used to obtain a reference MCNP solution for the whole UNF cask. Because of time limitation, the MCNP results near the cask boundaries have significant uncertainties. Except for these, the RAPID results are in excellent agreement with the MCNP predictions, and its computation time is significantly lower, 35 second on 1 core versus 9.5 days on 16 cores.

Comparative changes in color features and pigment composition of red wines aged in oak and cherry wood casks.

PubMed

Chinnici, Fabio; Natali, Nadia; Sonni, Francesca; Bellachioma, Attilio; Riponi, Claudio

2011-06-22

The color features and the evolution of both the monomeric and the derived pigments of red wines aged in oak and cherry 225 L barriques have been investigated during a four months period. For cherry wood, the utilization of 1000 L casks was tested as well. The use of cherry casks resulted in a faster evolution of pigments with a rapid decline of monomeric anthocyanins and a quick augmentation formation of derived and polymeric compounds. At the end of the aging, wines stored in oak and cherry barriques lost, respectively, about 20% and 80% of the initial pigment amount, while in the 1000 L cherry casks, the same compounds diminished by about 60%. Ethyl-bridged adducts and vitisins were the main class of derivatives formed, representing up to 25% of the total pigment amount in the cherry aged samples. Color density augmented in both the oak and cherry wood aged samples, but the latter had the highest values of this parameter. Because of the highly oxidative behavior of the cherry barriques, the use of larger casks (e.g., 1000 L) is proposed in the case of prolonged aging times.

Concrete Shield Performance of the VSC-17 Spent Nuclear Fuel Cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

Koji Shirai

2006-04-01

The VSC-17 Spent Nuclear Fuel Storage Cask was surveyed for degradation of the concrete shield by radiation measurement, temperature measurement, and ultrasonic testing. No general loss of shielding function was identified.

Effect of a dual-purpose cask payload increment of spent fuel assemblies from VVER 1000 Bushehr Nuclear Power Plant on basket criticality.

PubMed

Rezaeian, M; Kamali, J

2017-01-01

Dual-purpose casks can be utilized for dry interim storage and transportation of the highly radioactive spent fuel assemblies (SFAs) of Bushehr Nuclear Power Plant (NPP). Criticality safety analysis was carried out using the MCNP code for the cask containing 12, 18, or 19 SFAs. The basket materials of borated stainless steel and Boral (Al-B 4 C) were investigated, and the minimum required receptacle pitch of the basket was determined. Copyright © 2016 Elsevier Ltd. All rights reserved.

Thermal modeling of a vertical dry storage cask for used nuclear fuel

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Jie; Liu, Yung Y.

2016-05-01

Thermal modeling of temperature profiles of dry casks has been identified as a high-priority item in a U.S. Department of Energy gap analysis. In this work, a three-dimensional model of a vertical dry cask has been constructed for computer simulation by using the ANSYS/FLUENT code. The vertical storage cask contains a welded canister for 32 Pressurized Water Reactor (PWR) used-fuel assemblies with a total decay heat load of 34 kW. To simplify thermal calculations, an effective thermal conductivity model for a 17 x 17 PWR used (or spent)-fuel assembly was developed and used in the simulation of thermal performance. Themore » effects of canister fill gas (helium or nitrogen), internal pressure (1-6 atm), and basket material (stainless steel or aluminum alloy) were studied to determine the peak cladding temperature (PCT) and the canister surface temperatures (CSTs). The results showed that high thermal conductivity of the basket material greatly enhances heat transfer and reduces the PCT. The results also showed that natural convection affects both PCT and the CST profile, while the latter depends strongly on the type of fill gas and canister internal pressure. Of particular interest to condition and performance monitoring is the identification of canister locations where significant temperature change occurs after a canister is breached and the fill gas changes from high-pressure helium to ambient air. This study provided insight on the thermal performance of a vertical storage cask containing high-burnup fuel, and helped advance the concept of monitoring CSTs as a means to detect helium leakage from a welded canister. The effects of blockage of air inlet vents on the cask's thermal performance were studied. The simulation were validated by comparing the results against data obtained from the temperature measurements of a commercial cask.« less

Conceptual Design Report Cask Loadout Sys and Cask Drop Redesign for the Immersion Pail Support Structure and Operator Interface Platform at 105 K West

DOE Office of Scientific and Technical Information (OSTI.GOV)

LANGEVIN, A.S.

1999-07-12

This conceptual design report documents the redesign of the IPSS and the OIP in the 105 KW Basin south loadout pit due to a postulated cask drop accident, as part of Project A.5/A.6, Canister Transfer Facility Modifications. Project A.5/A.6 involves facility modifications needed to transfer fuel from the basin into the cask-MCO. The function of the IPSS is to suspend, guide, and position the immersion pail. The immersion pail protects the cask-MCO from contamination by basin water and acts as a lifting device for the cask-MCO. The OIP provides operator access to the south loadout pit. Previous analyses studied themore » effects of a cask-MCO drop on the south loadout pit concrete structure and on the IPSS. The most recent analysis considered the resulting loads at the pit slab/wall joint (Kanjilal, 1999). This area had not been modeled previously, and the analysis results indicate that the demand capacity exceeds the allowable at the slab/wall joint. The energy induced on the south loadout pit must be limited such that the safety class function of the basin is maintained. The solution presented in this CDR redesigns the IPSS and the OIP to include impact-absorbing features that will reduce the induced energy. The impact absorbing features of the new design include: Impact-absorbing material at the IPSS base and at the upper portion of the IPSS legs. A sleeve which provides a hydraulic means of absorbing energy. Designing the OIP to act as an impact absorber. The existing IPSS structure in 105 KW will be removed. This conceptual design considers only loads resulting from drops directly over the IPSS and south loadout pit area. Drops in other areas of the basin are not considered, and will be covered as part of a future revision to this CDR.« less

Radiation Templates of Spent Fuel in Casks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vanier, Peter

BNL and INL propose to perform a scoping study, using heavily collimated gamma and fast neutron detectors, to obtain passive radiation templates of dry storage casks containing spent fuel. The goal is to demonstrate sufficient spatial resolution and sensitivity to detect a missing fuel assembly. Such measurements, combined with detailed modeling and decay corrections should provide confidence that the cask contents have not been altered, despite loss of continuity of knowledge (CoK). The concept relies on the leakage of high energy gammas and neutrons through the shielding of the casks. Tests will emphasize organic scintillators with pulse shape discrimination, butmore » baseline comparisons will be made to high purity germanium (HPGe) and collimated moderated 3He detectors deployed in the same locations. Commercial off-the-shelf (COTS) detectors and data acquisition electronics will be used with custom-built collimators and shielding.« less

Spent fuel cask handling at an operating nuclear power plant

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pal, A.C.

1988-01-01

The importance of spent fuel handling at operating nuclear power plants cannot be overstated. Because of its highly radioactive nature, however, spent fuel must be handled in thick, lead-lined containers or casks. Thus, all casks for spent fuel handling are heavy loads by the US Nuclear Regulatory Commission's definition, and any load-drop must be evaluated for its potential to damage safety-related equipment. Nuclear Regulatory Guide NUREG-0612 prescribes the regulatory requirements of alternative heavy-load-handling methodologies such as (a) by providing cranes that meet the requirements of NUREG-0554, which shall be called the soft path, or (b) by providing protective devices atmore » all postulated load-drop areas to prevent any damage to safety-related equipment, which shall be called the hard path. The work reported in this paper relates to cask handling at New York Power Authority's James A. FitzPatrick (JAF) plant.« less

Modification and benchmarking of SKYSHINE-III for use with ISFSI cask arrays

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hertel, N.E.; Napolitano, D.G.

1997-12-01

Dry cask storage arrays are becoming more and more common at nuclear power plants in the United States. Title 10 of the Code of Federal Regulations, Part 72, limits doses at the controlled area boundary of these independent spent-fuel storage installations (ISFSI) to 0.25 mSv (25 mrem)/yr. The minimum controlled area boundaries of such a facility are determined by cask array dose calculations, which include direct radiation and radiation scattered by the atmosphere, also known as skyshine. NAC International (NAC) uses SKYSHINE-III to calculate the gamma-ray and neutron dose rates as a function of distance from ISFSI arrays. In thismore » paper, we present modifications to the SKYSHINE-III that more explicitly model cask arrays. In addition, we have benchmarked the radiation transport methods used in SKYSHINE-III against {sup 60}Co gamma-ray experiments and MCNP neutron calculations.« less

Development of Neutron Energy Spectral Signatures for Passive Monitoring of Spent Nuclear Fuels in Dry Cask Storage

NASA Astrophysics Data System (ADS)

Harkness, Ira; Zhu, Ting; Liang, Yinong; Rauch, Eric; Enqvist, Andreas; Jordan, Kelly A.

2018-01-01

Demand for spent nuclear fuel dry casks as an interim storage solution has increased globally and the IAEA has expressed a need for robust safeguards and verification technologies for ensuring the continuity of knowledge and the integrity of radioactive materials inside spent fuel casks. Existing research has been focusing on "fingerprinting" casks based on count rate statistics to represent radiation emission signatures. The current research aims to expand to include neutron energy spectral information as part of the fuel characteristics. First, spent fuel composition data are taken from the Next Generation Safeguards Initiative Spent Fuel Libraries, representative for Westinghouse 17ˣ17 PWR assemblies. The ORIGEN-S code then calculates the spontaneous fission and (α,n) emissions for individual fuel rods, followed by detailed MCNP simulations of neutrons transported through the fuel assemblies. A comprehensive database of neutron energy spectral profiles is to be constructed, with different enrichment, burn-up, and cooling time conditions. The end goal is to utilize the computational spent fuel library, predictive algorithm, and a pressurized 4He scintillator to verify the spent fuel assemblies inside a cask. This work identifies neutron spectral signatures that correlate with the cooling time of spent fuel. Both the total and relative contributions from spontaneous fission and (α,n) change noticeably with respect to cooling time, due to the relatively short half-life (18 years) of the major neutron source 244Cm. Identification of this and other neutron spectral signatures allows the characterization of spent nuclear fuels in dry cask storage.

PRELIMINARY REPORT: EFFECTS OF IRRADIATION AND THERMAL EXPOSURE ON ELASTOMERIC SEALS FOR CASK TRANSPORTATION AND STORAGE

DOE Office of Scientific and Technical Information (OSTI.GOV)

Verst, C.; Skidmore, E.; Daugherty, W.

2014-05-30

A testing and analysis approach to predict the sealing behavior of elastomeric seal materials in dry storage casks and evaluate their ability to maintain a seal under thermal and radiation exposure conditions of extended storage and beyond was developed, and initial tests have been conducted. The initial tests evaluate the aging response of EPDM elastomer O-ring seals. The thermal and radiation exposure conditions of the CASTOR® V/21 casks were selected for testing as this cask design is of interest due to its widespread use, and close proximity of the seals to the fuel compared to other cask designs leading tomore » a relatively high temperature and dose under storage conditions. A novel test fixture was developed to enable compression stress relaxation measurements for the seal material at the thermal and radiation exposure conditions. A loss of compression stress of 90% is suggested as the threshold at which sealing ability of an elastomeric seal would be lost. Previous studies have shown this value to be conservative to actual leakage failure for most aging conditions. These initial results indicate that the seal would be expected to retain sealing ability throughout extended storage at the cask design conditions, though longer exposure times are needed to validate this assumption. The high constant dose rate used in the testing is not prototypic of the decreasingly low dose rate that would occur under extended storage. The primary degradation mechanism of oxidation of polymeric compounds is highly dependent on temperature and time of exposure, and with radiation expected to exacerbate the oxidation.« less

75 FR 49813 - List of Approved Spent Fuel Storage Casks: MAGNASTOR System, Revision 1, Confirmation of...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-16

... Storage Casks: MAGNASTOR System, Revision 1, Confirmation of Effective Date AGENCY: Nuclear Regulatory... spent fuel storage regulations at 10 CFR 72.214 to revise the MAGNASTOR System listing to include...

75 FR 42339 - List of Approved Spent Fuel Storage Casks: NAC-MPC System, Revision 6

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-21

...; minor design modifications to the Vertical Concrete Cask (VCC) incorporating design features from the... (ALARA) principles; an increase in the concrete pad compression strength from 4000 psi to 6000 psi; added...

Fire resistant nuclear fuel cask

DOEpatents

Heckman, Richard C.; Moss, Marvin

1979-01-01

The disclosure is directed to a fire resistant nuclear fuel cask employing reversibly thermally expansible bands between adjacent cooling fins such that normal outward flow of heat is not interfered with, but abnormal inward flow of heat is impeded or blocked.

Palladium-Mediated Catalysis Leads to Intramolecular Narcissistic Self-Sorting on a Cavitand Platform.

PubMed

Nagymihály, Zoltán; Caturello, Naidel A M S; Takátsy, Anikó; Aragay, Gemma; Kollár, László; Albuquerque, Rodrigo Q; Csók, Zsolt

2017-01-06

Palladium-catalyzed aminocarbonylation reactions have been used to directly convert a tetraiodocavitand intermediate into the corresponding carboxamides and 2-ketocarboxamides. When complex mixtures of the amine reactants are employed in competition experiments using polar solvents, such as DMF, no "mixed" products possessing structurally different amide fragments are detected either by 1 H or 13 C NMR. Only highly symmetrical cavitands are sorted out of a large number of potentially feasible products, which represents a rare example of intramolecular, narcissistic self-sorting. Our experimental results along with thermodynamic energy analysis suggest that the observed self-sorting is a symmetry-driven, kinetically controlled process.

Personal Wellness Card Sort: A Strategy for Promoting Relational Healing

ERIC Educational Resources Information Center

Lenz, A. Stephen; Roscoe, Lauren J.

2011-01-01

The Personal Wellness Card Sort task is presented as a practical tool for mediating a therapeutic impasse when working with clients who have experienced a traumatic event. The authors describe how this activity can assist clients with viewing themselves in a holistic way, regaining disowned aspects of self and initiating movement toward connecting…

Amyloid-like aggregation of provasopressin in diabetes insipidus and secretory granule sorting.

PubMed

Beuret, Nicole; Hasler, Franziska; Prescianotto-Baschong, Cristina; Birk, Julia; Rutishauser, Jonas; Spiess, Martin

2017-01-26

Aggregation of peptide hormone precursors in the trans-Golgi network is an essential process in the biogenesis of secretory granules in endocrine cells. It has recently been proposed that this aggregation corresponds to the formation of functional amyloids. Our previous finding that dominant mutations in provasopressin, which cause cell degeneration and diabetes insipidus, prevent native folding and produce fibrillar aggregates in the endoplasmic reticulum (ER) might thus reflect mislocalized amyloid formation by sequences that evolved to mediate granule sorting. Here we identified two sequences responsible for fibrillar aggregation of mutant precursors in the ER: the N-terminal vasopressin nonapeptide and the C-terminal glycopeptide. To test their role in granule sorting, the glycopeptide was deleted and/or vasopressin mutated to inactivate ER aggregation while still permitting precursor folding and ER exit. These mutations strongly reduced sorting into granules and regulated secretion in endocrine AtT20 cells. The same sequences - vasopressin and the glycopeptide - mediate physiological aggregation of the wild-type hormone precursor into secretory granules and the pathological fibrillar aggregation of disease mutants in the ER. These findings support the amyloid hypothesis for secretory granule biogenesis.

ESCRT-II/Vps25 constrains digit number by endosome-mediated selective modulation of FGF-SHH signaling.

PubMed

Handschuh, Karen; Feenstra, Jennifer; Koss, Matthew; Ferretti, Elisabetta; Risolino, Maurizio; Zewdu, Rediet; Sahai, Michelle A; Bénazet, Jean-Denis; Peng, Xiao P; Depew, Michael J; Quintana, Laura; Sharpe, James; Wang, Baolin; Alcorn, Heather; Rivi, Roberta; Butcher, Stephen; Manak, J Robert; Vaccari, Thomas; Weinstein, Harel; Anderson, Kathryn V; Lacy, Elizabeth; Selleri, Licia

2014-10-23

Sorting and degradation of receptors and associated signaling molecules maintain homeostasis of conserved signaling pathways during cell specification and tissue development. Yet, whether machineries that sort signaling proteins act preferentially on different receptors and ligands in different contexts remains mysterious. Here, we show that Vacuolar protein sorting 25, Vps25, a component of ESCRT-II (Endosomal Sorting Complex Required for Transport II), directs preferential endosome-mediated modulation of FGF signaling in limbs. By ENU-induced mutagenesis, we isolated a polydactylous mouse line carrying a hypomorphic mutation of Vps25 (Vps25(ENU)). Unlike Vps25-null embryos we generated, Vps25(ENU/ENU) mutants survive until late gestation. Their limbs display FGF signaling enhancement and consequent hyperactivation of the FGF-SHH feedback loop causing polydactyly, whereas WNT and BMP signaling remain unperturbed. Notably, Vps25(ENU/ENU) Mouse Embryonic Fibroblasts exhibit aberrant FGFR trafficking and degradation; however, SHH signaling is unperturbed. These studies establish that the ESCRT-II machinery selectively limits FGF signaling in vertebrate skeletal patterning.

Microfluidic devices for label-free separation of cells through transient interaction with asymmetric receptor patterns

NASA Astrophysics Data System (ADS)

Bose, S.; Singh, R.; Hollatz, M. H.; Lee, C.-H.; Karp, J.; Karnik, R.

2012-02-01

Cell sorting serves an important role in clinical diagnosis and biological research. Most of the existing microscale sorting techniques are either non-specific to antigen type or rely on capturing cells making sample recovery difficult. We demonstrate a simple; yet effective technique for isolating cells in an antigen specific manner by using transient interactions of the cell surface antigens with asymmetric receptor patterned surface. Using microfluidic devices incorporating P-selectin patterns we demonstrate separation of HL60 cells from K562 cells. We achieved a sorting purity above 90% and efficiency greater than 85% with this system. We also present a mathematical model incorporating flow mediated and adhesion mediated transport of cells in the microchannel that can be used to predict the performance of these devices. Lastly, we demonstrate the clinical significance of the method by demonstrating single step separation of neutrophils from whole blood. When whole blood is introduced in the device, the granulocyte population gets separated exclusively yielding neutrophils of high purity (<10% RBC contamination). To our knowledge, this is the first ever demonstration of continuous label free sorting of neutrophils from whole blood. We believe this technology will be useful in developing point-of-care diagnostic devices and also for a host of cell sorting applications.

78 FR 78693 - List of Approved Spent Fuel Storage Casks: Transnuclear, Inc. Standardized NUHOMS® Cask System

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-27

... dangerous to living organisms, including insects, microbes, bacteria or virus that attach to dust that.... According to his comment, ``[T]he Deer Tick has carried a spirochete bacteria for millions of years, but...

DOE Office of Scientific and Technical Information (OSTI.GOV)

Evans, J.H.; Chipley, K.K.; Nelms, H.A.

An evaluation of the ORNL loop transport cask demonstrating its compliance with the regulations governing the transportation of radioactive and fissile materials is presented. A previous review of the cask is updated to demonstrate compliance with current regulations, to present current procedures, and to reflect the more recent technology.

Ageing of a neutron shielding used in transport/storage casks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nizeyiman, Fidele; Alami, Aatif; Issard, Herve

2012-07-11

In radioactive materials transport/storage casks, a mineral-filled vinylester composite is used for neutron shielding which relies on its hydrogen and boron atoms content. During cask service life, this composite is mainly subjected to three types of ageing: hydrothermal ageing, thermal oxidation and neutron irradiation. The aim of this study is to investigate the effect of hydrothermal ageing on the properties and chemical composition of this polymer composite. At high temperature (120 Degree-Sign C and 140 Degree-Sign C), the main consequence is the strong decrease of mechanical properties induced by the filler/matrix debonding.

A sorting nexin 17-binding domain within the LRP1 cytoplasmic tail mediates receptor recycling through the basolateral sorting endosome

PubMed Central

Farfán, Pamela; Lee, Jiyeon; Larios, Jorge; Sotelo, Pablo; Bu, Guojun; Marzolo, María-Paz

2013-01-01

Sorting nexin 17 (SNX17) is an adaptor protein present in EEA1-positive sorting endosomes that promotes the efficient recycling of low-density lipoprotein receptor-related protein 1 (LRP1) to the plasma membrane through recognition of the first NPxY motif in the cytoplasmic tail of this receptor. The interaction of LRP1 with SNX17 also regulates the basolateral recycling of the receptor from the basolateral sorting endosome (BSE). In contrast, megalin, which is apically distributed in polarized epithelial cells and localizes poorly to EEA1-positive sorting endosomes, does not interact with SNX17, despite containing three NPxY motifs, indicating that this motif is not sufficient for receptor recognition by SNX17. Here, we identified a cluster of 32 amino acids within the cytoplasmic domain of LRP1 that is both necessary and sufficient for SNX17 binding. To delineate the function of this SNX17-binding domain, we generated chimeric proteins in which the SNX17-binding domain was inserted into the cytoplasmic tail of megalin. This insertion mediated the binding of megalin to SNX17 and modified the cell surface expression and recycling of megalin in non-polarized cells. However, the polarized localization of chimeric megalin was not modified in polarized MDCK cells. These results provide evidence regarding the molecular and cellular mechanisms underlying the specificity of SNX17-binding receptors and the restricted function of SNX17 in the BSE. PMID:23593972

Sorting of the Neuroendocrine Secretory Protein Secretogranin II into the Regulated Secretory Pathway

PubMed Central

Courel, Maïté; Vasquez, Michael S.; Hook, Vivian Y.; Mahata, Sushil K.; Taupenot, Laurent

2008-01-01

Secretogranin II (SgII) belongs to the granin family of prohormones widely distributed in dense-core secretory granules (DCGs) of endocrine, neuroendocrine, and neuronal cells, including sympathoadrenal chromaffin cells. The mechanisms by which secretory proteins, and granins in particular, are sorted into the regulated secretory pathway are unsettled. We designed a strategy based on novel chimeric forms of human SgII fused to fluorescent (green fluorescent protein) or chemiluminescent (embryonic alkaline phosphatase) reporters to identify trafficking determinants mediating DCG targeting of SgII in sympathoadrenal cells. Three-dimensional deconvolution fluorescence microscopy and secretagogue-stimulated release studies demonstrate that SgII chimeras are correctly targeted to DCGs and released by exocytosis in PC12 and primary chromaffin cells. Results from a Golgi-retained mutant form of SgII suggest that sorting of SgII into DCGs depends on a saturable sorting machinery at the trans-Golgi/trans-Golgi network. Truncation analyses reveal the presence of DCG-targeting signals within both the N- and C-terminal regions of SgII, with the putative α-helix-containing SgII-(25-41) and SgII-(334-348) acting as sufficient, independent sorting domains. This study defines sequence features of SgII mediating vesicular targeting in sympathoadrenal cells and suggests a mechanism by which discrete domains of the molecule function in sorting, perhaps by virtue of a particular arrangement in tertiary structure and/or interaction with a specific component of the DCG membrane. PMID:18299326

Sorting of the neuroendocrine secretory protein Secretogranin II into the regulated secretory pathway: role of N- and C-terminal alpha-helical domains.

PubMed

Courel, Maïté; Vasquez, Michael S; Hook, Vivian Y; Mahata, Sushil K; Taupenot, Laurent

2008-04-25

Secretogranin II (SgII) belongs to the granin family of prohormones widely distributed in dense-core secretory granules (DCGs) of endocrine, neuroendocrine, and neuronal cells, including sympathoadrenal chromaffin cells. The mechanisms by which secretory proteins, and granins in particular, are sorted into the regulated secretory pathway are unsettled. We designed a strategy based on novel chimeric forms of human SgII fused to fluorescent (green fluorescent protein) or chemiluminescent (embryonic alkaline phosphatase) reporters to identify trafficking determinants mediating DCG targeting of SgII in sympathoadrenal cells. Three-dimensional deconvolution fluorescence microscopy and secretagogue-stimulated release studies demonstrate that SgII chimeras are correctly targeted to DCGs and released by exocytosis in PC12 and primary chromaffin cells. Results from a Golgi-retained mutant form of SgII suggest that sorting of SgII into DCGs depends on a saturable sorting machinery at the trans-Golgi/trans-Golgi network. Truncation analyses reveal the presence of DCG-targeting signals within both the N- and C-terminal regions of SgII, with the putative alpha-helix-containing SgII-(25-41) and SgII-(334-348) acting as sufficient, independent sorting domains. This study defines sequence features of SgII mediating vesicular targeting in sympathoadrenal cells and suggests a mechanism by which discrete domains of the molecule function in sorting, perhaps by virtue of a particular arrangement in tertiary structure and/or interaction with a specific component of the DCG membrane.

75 FR 36449 - Yankee Atomic Electric Co.; Yankee Atomic Independent Spent Fuel Storage Installation; Issuance...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-25

... Specification (TS) Surveillance Requirement 3.1.6.1 to verify the operability of the concrete cask heat removal....6.1 to verify the operability of the concrete cask heat removal system to maintain safe storage...

75 FR 34181 - Connecticut Yankee Atomic Power Company, Haddam Neck Plant, Independent Spent Fuel Storage...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-16

... Specification (TS) Surveillance Requirement 3.1.6.1 to verify the operability of the concrete cask heat removal... Specification (TS) Surveillance Requirement 3.1.6.1 to verify the operability of the concrete cask heat removal...

Endocytosis and Endosomal Trafficking in Plants.

PubMed

Paez Valencia, Julio; Goodman, Kaija; Otegui, Marisa S

2016-04-29

Endocytosis and endosomal trafficking are essential processes in cells that control the dynamics and turnover of plasma membrane proteins, such as receptors, transporters, and cell wall biosynthetic enzymes. Plasma membrane proteins (cargo) are internalized by endocytosis through clathrin-dependent or clathrin-independent mechanism and delivered to early endosomes. From the endosomes, cargo proteins are recycled back to the plasma membrane via different pathways, which rely on small GTPases and the retromer complex. Proteins that are targeted for degradation through ubiquitination are sorted into endosomal vesicles by the ESCRT (endosomal sorting complex required for transport) machinery for degradation in the vacuole. Endocytic and endosomal trafficking regulates many cellular, developmental, and physiological processes, including cellular polarization, hormone transport, metal ion homeostasis, cytokinesis, pathogen responses, and development. In this review, we discuss the mechanisms that mediate the recognition and sorting of endocytic and endosomal cargos, the vesiculation processes that mediate their trafficking, and their connection to cellular and physiological responses in plants.

High Burnup Dry Storage Cask Research and Development Project, Final Test Plan

DOE Office of Scientific and Technical Information (OSTI.GOV)

None

2014-02-27

EPRI is leading a project team to develop and implement the first five years of a Test Plan to collect data from a SNF dry storage system containing high burnup fuel.12 The Test Plan defined in this document outlines the data to be collected, and the storage system design, procedures, and licensing necessary to implement the Test Plan.13 The main goals of the proposed test are to provide confirmatory data14 for models, future SNF dry storage cask design, and to support license renewals and new licenses for ISFSIs. To provide data that is most relevant to high burnup fuel inmore » dry storage, the design of the test storage system must mimic real conditions that high burnup SNF experiences during all stages of dry storage: loading, cask drying, inert gas backfilling, and transfer to the ISFSI for multi-year storage.15 Along with other optional modeling, SETs, and SSTs, the data collected in this Test Plan can be used to evaluate the integrity of dry storage systems and the high burnup fuel contained therein over many decades. It should be noted that the Test Plan described in this document discusses essential activities that go beyond the first five years of Test Plan implementation.16 The first five years of the Test Plan include activities up through loading the cask, initiating the data collection, and beginning the long-term storage period at the ISFSI. The Test Plan encompasses the overall project that includes activities that may not be completed until 15 or more years from now, including continued data collection, shipment of the Research Project Cask to a Fuel Examination Facility, opening the cask at the Fuel Examination Facility, and examining the high burnup fuel after the initial storage period.« less

Full-Scale Accident Testing in Support of Used Nuclear Fuel Transportation.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Durbin, Samuel G.; Lindgren, Eric R.; Rechard, Rob P.

2014-09-01

The safe transport of spent nuclear fuel and high-level radioactive waste is an important aspect of the waste management system of the United States. The Nuclear Regulatory Commission (NRC) currently certifies spent nuclear fuel rail cask designs based primarily on numerical modeling of hypothetical accident conditions augmented with some small scale testing. However, NRC initiated a Package Performance Study (PPS) in 2001 to examine the response of full-scale rail casks in extreme transportation accidents. The objectives of PPS were to demonstrate the safety of transportation casks and to provide high-fidelity data for validating the modeling. Although work on the PPSmore » eventually stopped, the Blue Ribbon Commission on America’s Nuclear Future recommended in 2012 that the test plans be re-examined. This recommendation was in recognition of substantial public feedback calling for a full-scale severe accident test of a rail cask to verify evaluations by NRC, which find that risk from the transport of spent fuel in certified casks is extremely low. This report, which serves as the re-assessment, provides a summary of the history of the PPS planning, identifies the objectives and technical issues that drove the scope of the PPS, and presents a possible path for moving forward in planning to conduct a full-scale cask test. Because full-scale testing is expensive, the value of such testing on public perceptions and public acceptance is important. Consequently, the path forward starts with a public perception component followed by two additional components: accident simulation and first responder training. The proposed path forward presents a series of study options with several points where the package performance study could be redirected if warranted.« less

Impact Analyses and Tests of Metal Cask Considering Aircraft Engine Crash - 12308

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lee, Sanghoon; Choi, Woo-Seok; Kim, Ki-Young

2012-07-01

The structural integrity of a dual purpose metal cask currently under development by the Korea Radioactive Waste Management Cooperation (KRMC) is evaluated through analyses and tests under a high-speed missile impact considering the targeted aircraft crash conditions. The impact conditions were carefully chosen through a survey on accident cases and recommendations from the literature. The missile impact velocity was set at 150 m/s, and two impact orientations were considered. A simplified missile simulating a commercial aircraft engine is designed from an impact load history curve provided in the literature. In the analyses, the focus is on the evaluation of themore » containment boundary integrity of the metal cask. The analyses results are compared with the results of tests using a 1/3 scale model. The results show very good agreements, and the procedure and methodology adopted in the structural analyses are validated. While the integrity of the cask is maintained in one evaluation where the missile impacts the top side of the free standing cask, the containment boundary is breached in another case in which the missile impacts the center of the cask lid in a perpendicular orientation. A safety assessment using a numerical simulation of an aircraft engine crash into spent nuclear fuel storage systems is performed. A commercially available explicit finite element code is utilized for the dynamic simulation, and the strain rate effect is included in the modeling of the materials used in the target system and missile. The simulation results show very good agreement with the test results. It is noted that this is the first test considering an aircraft crash in Korea. (authors)« less

Multiple motifs regulate apical sorting of p75 via a mechanism that involves dimerization and higher-order oligomerization

PubMed Central

Youker, Robert T.; Bruns, Jennifer R.; Costa, Simone A.; Rbaibi, Youssef; Lanni, Frederick; Kashlan, Ossama B.; Teng, Haibing; Weisz, Ora A.

2013-01-01

The sorting signals that direct proteins to the apical surface of polarized epithelial cells are complex and can include posttranslational modifications, such as N- and O-linked glycosylation. Efficient apical sorting of the neurotrophin receptor p75 is dependent on its O-glycosylated membrane proximal stalk, but how this domain mediates targeting is unknown. Protein oligomerization or clustering has been suggested as a common step in the segregation of all apical proteins. Like many apical proteins, p75 forms dimers, and we hypothesized that formation of higher-order clusters mediated by p75 dimerization and interactions of the stalk facilitate its apical sorting. Using fluorescence fluctuation techniques (photon-counting histogram and number and brightness analyses) to study p75 oligomerization status in vivo, we found that wild-type p75–green fluorescent protein forms clusters in the trans-Golgi network (TGN) but not at the plasma membrane. Disruption of either the dimerization motif or the stalk domain impaired both clustering and polarized delivery. Manipulation of O-glycan processing or depletion of multiple galectins expressed in Madin-Darby canine kidney cells had no effect on p75 sorting, suggesting that the stalk domain functions as a structural prop to position other determinants in the lumenal domain of p75 for oligomerization. Additionally, a p75 mutant with intact dimerization and stalk motifs but with a dominant basolateral sorting determinant (Δ250 mutant) did not form oligomers, consistent with a requirement for clustering in apical sorting. Artificially enhancing dimerization restored clustering to the Δ250 mutant but was insufficient to reroute this mutant to the apical surface. Together these studies demonstrate that clustering in the TGN is required for normal biosynthetic apical sorting of p75 but is not by itself sufficient to reroute a protein to the apical surface in the presence of a strong basolateral sorting determinant. Our studies shed new light on the hierarchy of polarized sorting signals and on the mechanisms by which newly synthesized proteins are segregated in the TGN for eventual apical delivery. PMID:23637462

Used fuel extended storage security and safeguards by design roadmap

DOE Office of Scientific and Technical Information (OSTI.GOV)

Durbin, Samuel G.; Lindgren, Eric Richard; Jones, Robert

2016-05-01

In the United States, spent nuclear fuel (SNF) is safely and securely stored in spent fuel pools and dry storage casks. The available capacity in spent fuel pools across the nuclear fleet has nearly reached a steady state value. The excess SNF continues to be loaded in dry storage casks. Fuel is expected to remain in dry storage for periods beyond the initial dry cask certification period of 20 years. Recent licensing renewals have approved an additional 40 years. This report identifies the current requirements and evaluation techniques associated with the safeguards and security of SNF dry cask storage. Amore » set of knowledge gaps is identified in the current approaches. Finally, this roadmap identifies known knowledge gaps and provides a research path to deliver the tools and models needed to close the gaps and allow the optimization of the security and safeguards approaches for an interim spent fuel facility over the lifetime of the storage site.« less

Feasibility study for a transportation operations system cask maintenance facility

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rennich, M.J.; Medley, L.G.; Attaway, C.R.

The US Department of Energy (DOE), Office of Civilian Radioactive Waste Management (OCRWM) is responsible for the development of a waste management program for the disposition of spent nuclear fuel (SNF) and high-level waste (HLW). The program will include a transportation system for moving the nuclear waste from the sources to a geologic repository for permanent disposal. Specially designed casks will be used to safely transport the waste. The cask systems must be operated within limits imposed by DOE, the Nuclear Regulatory Commission (NRC), and the Department of Transportation (DOT). A dedicated facility for inspecting, testing, and maintaining the caskmore » systems was recommended by the General Accounting Office (in 1979) as the best means of assuring their operational effectiveness and safety, as well as regulatory compliance. In November of 1987, OCRWM requested a feasibility study be made of a Cask Maintenance Facility (CMF) that would perform the required functions. 46 refs., 16 figs., 13 tabs.« less

Analysis of Corrosion Residues Collected from the Aluminum Basket Rails of the High-Burnup Demonstration Cask.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bryan, Charles R.

On September, 2015, an inspection was performed on the TN-32B cask that will be used for the high-burnup demonstration project. During the survey, wooden cribbing that had been placed within the cask eleven years earlier to prevent shifting of the basket during transport was removed, revealing two areas of residue on the aluminum basket rails, where they had contacted the cribbing. The residue appeared to be a corrosion product, and concerns were raised that similar attack could exist at more difficult-to-inspect locations in the canister. Accordingly, when the canister was reopened, samples of the residue were collected for analysis. Thismore » report presents the results of that assessment, which determined that the corrosion was due to the presence of the cribbing. The corrosion was associated with fungal material, and fungal activity likely contributed to an aggressive chemical environment. Once the cask has been cleaned, there will be no risk of further corrosion.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ibarra, Luis; Sanders, David; Yang, Haori

The main goal of this study is to evaluate the long-term seismic performance of freestanding and anchored Dry Storage Casks (DSCs) using experimental tests on a shaking table, as well as comprehensive numerical evaluations that include the cask-pad-soil system. The study focuses on the dynamic performance of vertical DSCs, which can be designed as free-standing structures resting on a reinforced concrete foundation pad, or casks anchored to a foundation pad. The spent nuclear fuel (SNF) at nuclear power plants (NPPs) is initially stored in fuel-storage pools to control the fuel temperature. After several years, the fuel assemblies are transferred tomore » DSCs at sites contiguous to the plant, known as Interim Spent Fuel Storage Installations (ISFSIs). The regulations for these storage systems (10 CFR 72) ensure adequate passive heat removal and radiation shielding during normal operations, off-normal events, and accident scenarios. The integrity of the DSCs is important, even if the overpack does not breach, because eventually the spent fuel-rods need to be shipped either to a reprocessing plant or a repository. DSCs have been considered as a temporary storage solution, and usually are licensed for 20 years, although they can be relicensed for operating periods of up to 60 years. In recent years, DSCs have been reevaluated as a potential mid-term solution, in which the operating period may be extended for up to 300 years. At the same time, recent seismic events have underlined the significant risks DSCs are exposed. The consideration of DCSs for storing spent fuel for hundreds of years has created new challenges. In the case of seismic hazard, longer-term operating periods not only lead to larger horizontal accelerations, but also increase the relative effect of vertical accelerations that usually are disregarded for smaller seismic events. These larger seismic demands could lead to casks sliding and tipping over, impacting the concrete pad or adjacent casks. The casks may also slide and collide with other casks or structural components. Also, the different DSC components may impact each other during these events. This study provides a comprehensive evaluation of DSCs subjected to these extreme demands, including the effect of vertical accelerations, and soilstructure interaction.« less

GGA1 regulates signal-dependent sorting of BACE1 to recycling endosomes, which moderates Aβ production

PubMed Central

Toh, Wei Hong; Chia, Pei Zhi Cheryl; Hossain, Mohammed Iqbal; Gleeson, Paul A.

2018-01-01

The diversion of the membrane-bound β-site amyloid precursor protein–(APP) cleaving enzyme (BACE1) from the endolysosomal pathway to recycling endosomes represents an important transport step in the regulation of amyloid beta (Aβ) production. However, the mechanisms that regulate endosome sorting of BACE1 are poorly understood. Here we assessed the transport of BACE1 from early to recycling endosomes and have identified essential roles for the sorting nexin 4 (SNX4)-mediated, signal-independent pathway and for a novel signal-mediated pathway. The signal-mediated pathway is regulated by the phosphorylation of the DXXLL-motif sequence DISLL in the cytoplasmic tail of BACE1. The phosphomimetic S498D BACE1 mutant was trafficked to recycling endosomes at a faster rate compared with wild-type BACE1 or the nonphosphorylatable S498A mutant. The rapid transit of BACE1 S498D from early endosomes was coupled with reduced levels of amyloid precursor protein processing and Aβ production, compared with the S498A mutant. We show that the adaptor, GGA1, and retromer are essential to mediate rapid trafficking of phosphorylated BACE1 to recycling endosomes. In addition, the BACE1 DISLL motif is phosphorylated and regulates endosomal trafficking, in primary neurons. Therefore, post-translational phosphorylation of DISLL enhances the exit of BACE1 from early endosomes, a pathway mediated by GGA1 and retromer, which is important in regulating Aβ production. PMID:29142073

DOE Office of Scientific and Technical Information (OSTI.GOV)

Winston, Philip Lon

Prior to performing an internal visual inspection, samples of the headspace gas of the GNS Castor V/21 cask were taken on June 12, 2014. These samples were taken in support of the CREIPI/Japanese nuclear industry effort to validate fuel integrity without visual inspection by measuring the 85Kr content of the cask headspace

78 FR 16601 - List of Approved Spent Fuel Storage Casks: MAGNASTOR® System

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-18

... Storage Casks: MAGNASTOR[supreg] System AGENCY: Nuclear Regulatory Commission. ACTION: Direct final rule. SUMMARY: The U.S. Nuclear Regulatory Commission (NRC) is amending its spent fuel storage regulations by revising the NAC International, Inc. (NAC) Modular Advanced Generation Nuclear All-purpose Storage...

78 FR 32077 - List of Approved Spent Fuel Storage Casks: MAGNASTOR® System

Federal Register 2010, 2011, 2012, 2013, 2014

2013-05-29

... Fuel Storage Casks: MAGNASTOR[supreg] System AGENCY: Nuclear Regulatory Commission. ACTION: Direct... final rule that would have revised its spent fuel storage regulations to include Amendment No. 3 to... All-purpose Storage (MAGNASTOR[supreg]) System listing within the ``List of Approved Spent Fuel...

Ultrasonic Fingerprinting of Structural Materials: Spent Nuclear Fuel Containers Case-Study

NASA Astrophysics Data System (ADS)

Sednev, D.; Lider, A.; Demyanuk, D.; Kroening, M.; Salchak, Y.

Nowadays, NDT is mainly focused on safety purposes, but it seems possible to apply those methods to provide national and IAEA safeguards. The containment of spent fuel in storage casks could be dramatically improved in case of development of so-called "smart" spent fuel storage and transfer casks. Such casks would have tamper indicating and monitoring/tracking features integrated directly into the cask design. The microstructure of the containers material as well as of the dedicated weld seam is applied to the lid and the cask body and provides a unique fingerprint of the full container, which can be reproducibly scanned by using an appropriate technique. The echo-sounder technique, which is the most commonly used method for material inspection, was chosen for this project. The main measuring parameter is acoustic noise, reflected from material's artefacts. The purpose is to obtain structural fingerprinting. Reference measurement and additional measurement results were compared. Obtained results have verified the appliance of structural fingerprint and the chosen control method. The successful authentication demonstrates the levels of the feature points' compliance exceeding the given threshold which differs considerably from the percentage of the concurrent points during authentication from other points. Since reproduction or doubling of the proposed unique identification characteristics is impossible at the current state science and technology, application of this technique is considered to identify the interference into the nuclear materials displacement with high accuracy.

10 CFR 72.230 - Procedures for spent fuel storage cask submittals.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 2 2010-01-01 2010-01-01 false Procedures for spent fuel storage cask submittals. 72.230 Section 72.230 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSING REQUIREMENTS FOR THE INDEPENDENT STORAGE OF SPENT NUCLEAR FUEL, HIGH-LEVEL RADIOACTIVE WASTE, AND REACTOR-RELATED GREATER THAN CLASS C...

10 CFR 72.240 - Conditions for spent fuel storage cask reapproval.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 2 2010-01-01 2010-01-01 false Conditions for spent fuel storage cask reapproval. 72.240 Section 72.240 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSING REQUIREMENTS FOR THE INDEPENDENT STORAGE OF SPENT NUCLEAR FUEL, HIGH-LEVEL RADIOACTIVE WASTE, AND REACTOR-RELATED GREATER THAN CLASS C...

77 FR 64834 - Computational Fluid Dynamics Best Practice Guidelines for Dry Cask Applications

Federal Register 2010, 2011, 2012, 2013, 2014

2012-10-23

... NUCLEAR REGULATORY COMMISSION [NRC-2012-0250] Computational Fluid Dynamics Best Practice... public comments on draft NUREG-2152, ``Computational Fluid Dynamics Best Practice Guidelines for Dry Cask... System (ADAMS): You may access publicly-available documents online in the NRC Library at http://www.nrc...

77 FR 26050 - Burnup Credit in the Criticality Safety Analyses of Pressurized Water Reactor Spent Fuel in...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-02

... Pressurized Water Reactor Spent Fuel in Transportation and Storage Casks AGENCY: Nuclear Regulatory Commission... of pressurized water reactor spent nuclear fuel (SNF) in transportation packages and storage casks... for the licensing basis, (b) provide recommendations regarding advanced isotopic depletion and...

A review of ventilated storage cask (VSC) system projects and experience

DOE Office of Scientific and Technical Information (OSTI.GOV)

McConaghy, W.

1995-12-31

First, the author discusses the ventilated storage cask (VSC) design and an operations summary is given. Next VSC project status at Palisades, Point Beach, Arkansas Nuclear One, Fast Flux Test Facility and Zaporozhye is discussed. Lastly, VSC operational experience and VSC transportation interfaces are reviewed.

10 CFR 72.214 - List of approved spent fuel storage casks.

Code of Federal Regulations, 2012 CFR

2012-01-01

... specified in their Certificates of Compliance. Certificate Number: 1000 SAR Submitted by: General Nuclear Systems, Inc. SAR Title: Topical Safety Analysis Report for the Castor V/21 Cask Independent Spent Fuel... Model Number: CASTOR V/21 Certificate Number: 1002 SAR Submitted by: Nuclear Assurance Corporation SAR...

10 CFR 72.214 - List of approved spent fuel storage casks.

Code of Federal Regulations, 2014 CFR

2014-01-01

... specified in their Certificates of Compliance. Certificate Number: 1000 SAR Submitted by: General Nuclear Systems, Inc. SAR Title: Topical Safety Analysis Report for the Castor V/21 Cask Independent Spent Fuel... Model Number: CASTOR V/21 Certificate Number: 1002 SAR Submitted by: Nuclear Assurance Corporation SAR...

10 CFR 72.214 - List of approved spent fuel storage casks.

Code of Federal Regulations, 2013 CFR

2013-01-01

... specified in their Certificates of Compliance. Certificate Number: 1000 SAR Submitted by: General Nuclear Systems, Inc. SAR Title: Topical Safety Analysis Report for the Castor V/21 Cask Independent Spent Fuel... Model Number: CASTOR V/21 Certificate Number: 1002 SAR Submitted by: Nuclear Assurance Corporation SAR...

A sorting nexin 17-binding domain within the LRP1 cytoplasmic tail mediates receptor recycling through the basolateral sorting endosome.

PubMed

Farfán, Pamela; Lee, Jiyeon; Larios, Jorge; Sotelo, Pablo; Bu, Guojun; Marzolo, María-Paz

2013-07-01

Sorting nexin 17 (SNX17) is an adaptor protein present in early endosomal antigen 1 (EEA1)-positive sorting endosomes that promotes the efficient recycling of low-density lipoprotein receptor-related protein 1 (LRP1) to the plasma membrane through recognition of the first NPxY motif in the cytoplasmic tail of this receptor. The interaction of LRP1 with SNX17 also regulates the basolateral recycling of the receptor from the basolateral sorting endosome (BSE). In contrast, megalin, which is apically distributed in polarized epithelial cells and localizes poorly to EEA1-positive sorting endosomes, does not interact with SNX17, despite containing three NPxY motifs, indicating that this motif is not sufficient for receptor recognition by SNX17. Here, we identified a cluster of 32 amino acids within the cytoplasmic domain of LRP1 that is both necessary and sufficient for SNX17 binding. To delineate the function of this SNX17-binding domain, we generated chimeric proteins in which the SNX17-binding domain was inserted into the cytoplasmic tail of megalin. This insertion mediated the binding of megalin to SNX17 and modified the cell surface expression and recycling of megalin in non-polarized cells. However, the polarized localization of chimeric megalin was not modified in polarized Madin-Darby canine kidney cells. These results provide evidence regarding the molecular and cellular mechanisms underlying the specificity of SNX17-binding receptors and the restricted function of SNX17 in the BSE. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

UBE4B Protein Couples Ubiquitination and Sorting Machineries to Enable Epidermal Growth Factor Receptor (EGFR) Degradation*

PubMed Central

Sirisaengtaksin, Natalie; Gireud, Monica; Yan, Qing; Kubota, Yoshihisa; Meza, Denisse; Waymire, Jack C.; Zage, Peter E.; Bean, Andrew J.

2014-01-01

The signaling of plasma membrane proteins is tuned by internalization and sorting in the endocytic pathway prior to recycling or degradation in lysosomes. Ubiquitin modification allows recognition and association of cargo with endosomally associated protein complexes, enabling sorting of proteins to be degraded from those to be recycled. The mechanism that provides coordination between the cellular machineries that mediate ubiquitination and endosomal sorting is unknown. We report that the ubiquitin ligase UBE4B is recruited to endosomes in response to epidermal growth factor receptor (EGFR) activation by binding to Hrs, a key component of endosomal sorting complex required for transport (ESCRT) 0. We identify the EGFR as a substrate for UBE4B, establish UBE4B as a regulator of EGFR degradation, and describe a mechanism by which UBE4B regulates endosomal sorting, affecting cellular levels of the EGFR and its downstream signaling. We propose a model in which the coordinated action of UBE4B, ESCRT-0, and the deubiquitinating enzyme USP8 enable the endosomal sorting and lysosomal degradation of the EGFR. PMID:24344129

Benchmarking Data for the Proposed Signature of Used Fuel Casks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rauch, Eric Benton

2016-09-23

A set of benchmarking measurements to test facets of the proposed extended storage signature was conducted on May 17, 2016. The measurements were designed to test the overall concept of how the proposed signature can be used to identify a used fuel cask based only on the distribution of neutron sources within the cask. To simulate the distribution, 4 Cf-252 sources were chosen and arranged on a 3x3 grid in 3 different patterns and raw neutron totals counts were taken at 6 locations around the grid. This is a very simplified test of the typical geometry studied previously in simulationmore » with simulated used nuclear fuel.« less

77 FR 24585 - List of Approved Spent Fuel Storage Casks: HI-STORM 100, Revision 8

Federal Register 2010, 2011, 2012, 2013, 2014

2012-04-25

... Storage Casks: HI-STORM 100, Revision 8 AGENCY: Nuclear Regulatory Commission. ACTION: Direct final rule... revising the Holtec International HI-STORM 100 System listing within the ``List of Approved Spent Fuel...) 72.214, by revising the Holtec International HI-STORM 100 System listing within the ``List of...

77 FR 60479 - Burnup Credit in the Criticality Safety Analyses of Pressurized Water Reactor Spent Fuel in...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-10-03

... Pressurized Water Reactor Spent Fuel in Transportation and Storage Casks AGENCY: Nuclear Regulatory Commission... 3, entitled, ``Burnup Credit in the Criticality Safety Analyses of PWR [Pressurized Water Reactor... water reactor spent nuclear fuel (SNF) in transportation packages and storage casks. SFST-ISG-8...

10 CFR 72.240 - Conditions for spent fuel storage cask renewal.

Code of Federal Regulations, 2013 CFR

2013-01-01

... to exceed 40 years. In the event that the certificate holder does not apply for a cask design renewal...) The application must be accompanied by a safety analysis report (SAR). The SAR must include the following: (1) Design bases information as documented in the most recently updated final safety analysis...

10 CFR 72.240 - Conditions for spent fuel storage cask renewal.

Code of Federal Regulations, 2012 CFR

2012-01-01

... to exceed 40 years. In the event that the certificate holder does not apply for a cask design renewal...) The application must be accompanied by a safety analysis report (SAR). The SAR must include the following: (1) Design bases information as documented in the most recently updated final safety analysis...

10 CFR 72.240 - Conditions for spent fuel storage cask renewal.

Code of Federal Regulations, 2014 CFR

2014-01-01

... to exceed 40 years. In the event that the certificate holder does not apply for a cask design renewal...) The application must be accompanied by a safety analysis report (SAR). The SAR must include the following: (1) Design bases information as documented in the most recently updated final safety analysis...

Translation and evaluation of the Cultural Awareness Scale for Korean nursing students.

PubMed

Oh, Hyunjin; Lee, Jung-ah; Schepp, Karen G

2015-02-20

To evaluate the effectiveness of a curriculum for achieving high levels of cultural competence, we need to be able to assess education intended to enhance cultural competency skills. We therefore translated the Cultural Awareness Scale (CAS) into Korean (CAS-K). The purpose of this study was to evaluate the cross-cultural applicability and psychometric properties of the CAS-K, specifically its reliability and validity. A cross-sectional descriptive design was used to conduct the evaluation. A convenience sample of 495 nursing students was recruited from four levels of nursing education within four universities in the city of Daejeon, South Korea. This study provided beginning evidence of the validity and reliability of the CAS-K and the cross-cultural applicability of the concepts underlying this instrument. Cronbach's alpha ranged between 0.59 and 0.86 (overall 0.89) in the tests of internal consistency. Cultural competency score prediction of the experience of travel abroad (r=0.084) and the perceived need for cultural education (r=0.223) suggested reasonable criterion validity. Five factors with eigenvalues >1.0 were extracted, accounting for 55.58% of the variance; two retained the same items previously identified for the CAS. The CAS-K demonstrated satisfactory validity and reliability in measuring cultural awareness in this sample of Korean nursing students. The revised CAS-K should be tested for its usability in curriculum evaluation and its applicability as a guide for teaching cultural awareness among groups of Korean nursing students.

Detection of Missing Assemblies and Estimation of the Scattering Densities in a VSC-24 Dry Storage Cask with Cosmic-Ray-Muon-Based Computed Tomography

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Zhengzhi; Hayward, Jason; Liao, Can

We report that highly energetic, cosmic-ray muons can penetrate a dry storage cask and yield information about the material inside it by making use of the physics of multiple Coulomb scattering. Work by others has shown this information may be used for verification of dry storage cask contents after continuity of knowledge has been lost. In our modeling and simulation approach, we use ideal planar radiation detectors to record the trajectories and momentum of both incident and exiting cosmic ray muons; this choice allows us to demonstrate the fundamental limit of the technology for a particular measurement and reconstruction method.more » In a method analogous to computed tomography with the attenuation coefficient replaced by scattering density, we apply a filtered back projection algorithm in order to reconstruct the geometry in modeled scenarios for a VSC-24 concrete-walled cask. We also report on our attempt to estimate material-specific information. A scenario where one of the middle four spent nuclear fuel assemblies is missing—undetectable with a simple PoCA-based approach—is expected to be detectable with a CT-based approach. Moreover, a trickier scenario where one or more assemblies is replaced by a dummy assembly is put forward. Lastly, in this case, we expect that this dry storage cask should be found to be not as declared based on our simulation and reconstruction results.« less

Detection of Missing Assemblies and Estimation of the Scattering Densities in a VSC-24 Dry Storage Cask with Cosmic-Ray-Muon-Based Computed Tomography

DOE PAGES

Liu, Zhengzhi; Hayward, Jason; Liao, Can; ...

2017-08-01

We report that highly energetic, cosmic-ray muons can penetrate a dry storage cask and yield information about the material inside it by making use of the physics of multiple Coulomb scattering. Work by others has shown this information may be used for verification of dry storage cask contents after continuity of knowledge has been lost. In our modeling and simulation approach, we use ideal planar radiation detectors to record the trajectories and momentum of both incident and exiting cosmic ray muons; this choice allows us to demonstrate the fundamental limit of the technology for a particular measurement and reconstruction method.more » In a method analogous to computed tomography with the attenuation coefficient replaced by scattering density, we apply a filtered back projection algorithm in order to reconstruct the geometry in modeled scenarios for a VSC-24 concrete-walled cask. We also report on our attempt to estimate material-specific information. A scenario where one of the middle four spent nuclear fuel assemblies is missing—undetectable with a simple PoCA-based approach—is expected to be detectable with a CT-based approach. Moreover, a trickier scenario where one or more assemblies is replaced by a dummy assembly is put forward. Lastly, in this case, we expect that this dry storage cask should be found to be not as declared based on our simulation and reconstruction results.« less

CVAK104 is a Novel Regulator of Clathrin-mediated SNARE Sorting

PubMed Central

Borner, Georg H H; Rana, Amer A; Forster, Rebecca; Harbour, Michael; Smith, James C; Robinson, Margaret S

2007-01-01

Clathrin-coated vesicles (CCVs) mediate transport between the plasma membrane, endosomes and the trans Golgi network. Using comparative proteomics, we have identified coated-vesicle-associated kinase of 104 kDa (CVAK104) as a candidate accessory protein for CCV-mediated trafficking. Here, we demonstrate that the protein colocalizes with clathrin and adaptor protein-1 (AP-1), and that it is associated with a transferrin-positive endosomal compartment. Consistent with these observations, clathrin as well as the cargo adaptors AP-1 and epsinR can be coimmunoprecipitated with CVAK104. Small interfering RNA (siRNA) knockdown of CVAK104 in HeLa cells results in selective loss of the SNARE proteins syntaxin 8 and vti1b from CCVs. Morpholino-mediated knockdown of CVAK104 in Xenopus tropicalis causes severe developmental defects, including a bent body axis and ventral oedema. Thus, CVAK104 is an evolutionarily conserved protein involved in SNARE sorting that is essential for normal embryonic development. PMID:17587408

Smart Winery: A Real-Time Monitoring System for Structural Health and Ullage in Fino Style Wine Casks

PubMed Central

Cañete, Eduardo; Chen, Jaime; Rubio, Bartolomé

2018-01-01

The rapid development in low-cost sensor and wireless communication technology has made it possible for a large number of devices to coexist and exchange information autonomously. It has been predicted that a substantial number of devices will be able to exchange and provide information about an environment with the goal of improving our lives, under the well-known paradigm of the Internet of Things (IoT). One of the main applications of these kinds of devices is the monitoring of scenarios. In order to improve the current wine elaboration process, this paper presents a real-time monitoring system to supervise the status of wine casks. We have focused on a special kind of white wine, called Fino, principally produced in Andalusia (Southern Spain). The process by which this kind of wind is monitored is completely different from that of red wine, as the casks are not completely full and, due to the fact that they are not renewed very often, are more prone to breakage. A smart cork prototype monitors the structural health, the ullage, and the level of light inside the cask and the room temperature. The advantage of this smart cork is that it allows winemakers to monitor, in real time, the status of each wine cask so that, if an issue is detected (e.g., a crack appears in the cask), they can act immediately to resolve it. Moreover, abnormal parameters or incorrect environmental conditions can be detected in time before the wine loses its desired qualities. The system has been tested in “Bodegas San Acacio,” a winery based in Montemayor, a town in the north of Andalusia. Results show that the use of such a system can provide a solution that tracks the evolution and assesses the suitability of the delicate wine elaboration process in real time, which is especially important for the kind of wine considered in this paper. PMID:29518928

Spent nuclear fuel integrity during dry storage - performance tests and demonstrations

DOE Office of Scientific and Technical Information (OSTI.GOV)

McKinnon, M.A.; Doherty, A.L.

1997-06-01

This report summarizes the results of fuel integrity surveillance determined from gas sampling during and after performance tests and demonstrations conducted from 1983 through 1996 by or in cooperation with the US DOE Office of Commercial Radioactive Waste Management (OCRWM). The cask performance tests were conducted at Idaho National Engineering Laboratory (INEL) between 1984 and 1991 and included visual observation and ultrasonic examination of the condition of the cladding, fuel rods, and fuel assembly hardware before dry storage and consolidation of fuel, and a qualitative determination of the effects of dry storage and fuel consolidation on fission gas release frommore » the spent fuel rods. The performance tests consisted of 6 to 14 runs involving one or two loading, usually three backfill environments (helium, nitrogen, and vacuum backfills), and one or two storage system orientations. The nitrogen and helium backfills were sampled and analyzed to detect leaking spent fuel rods. At the end of each performance test, periodic gas sampling was conducted on each cask. A spent fuel behavior project (i.e., enhanced surveillance, monitoring, and gas sampling activities) was initiated by DOE in 1994 for intact fuel in a CASTOR V/21 cask and for consolidated fuel in a VSC-17 cask. The results of the gas sampling activities are included in this report. Information on spent fuel integrity is of interest in evaluating the impact of long-term dry storage on the behavior of spent fuel rods. Spent fuel used during cask performance tests at INEL offers significant opportunities for confirmation of the benign nature of long-term dry storage. Supporting cask demonstration included licensing and operation of an independent spent fuel storage installation (ISFSI) at the Virginia Power (VP) Surry reactor site. A CASTOR V/21, an MC-10, and a Nuclear Assurance NAC-I28 have been loaded and placed at the VP ISFSI as part of the demonstration program. 13 refs., 14 figs., 9 tabs.« less

Smart Winery: A Real-Time Monitoring System for Structural Health and Ullage in Fino Style Wine Casks.

PubMed

Cañete, Eduardo; Chen, Jaime; Martín, Cristian; Rubio, Bartolomé

2018-03-07

The rapid development in low-cost sensor and wireless communication technology has made it possible for a large number of devices to coexist and exchange information autonomously. It has been predicted that a substantial number of devices will be able to exchange and provide information about an environment with the goal of improving our lives, under the well-known paradigm of the Internet of Things (IoT). One of the main applications of these kinds of devices is the monitoring of scenarios. In order to improve the current wine elaboration process, this paper presents a real-time monitoring system to supervise the status of wine casks. We have focused on a special kind of white wine, called Fino, principally produced in Andalusia (Southern Spain). The process by which this kind of wind is monitored is completely different from that of red wine, as the casks are not completely full and, due to the fact that they are not renewed very often, are more prone to breakage. A smart cork prototype monitors the structural health, the ullage, and the level of light inside the cask and the room temperature. The advantage of this smart cork is that it allows winemakers to monitor, in real time, the status of each wine cask so that, if an issue is detected (e.g., a crack appears in the cask), they can act immediately to resolve it. Moreover, abnormal parameters or incorrect environmental conditions can be detected in time before the wine loses its desired qualities. The system has been tested in "Bodegas San Acacio," a winery based in Montemayor, a town in the north of Andalusia. Results show that the use of such a system can provide a solution that tracks the evolution and assesses the suitability of the delicate wine elaboration process in real time, which is especially important for the kind of wine considered in this paper.

78 FR 63408 - List of Approved Spent Fuel Storage Casks: Transnuclear, Inc. Standardized NUHOMS® Cask System

Federal Register 2010, 2011, 2012, 2013, 2014

2013-10-24

...: Gregory R. Trussell, Office of Federal and State Materials and Environmental Management Programs, U.S... Access and Management System (ADAMS): You may access publicly-available documents online in the NRC... continues to be ensured. The direct final rule will become effective on January 7, 2014. However, if the NRC...

A&M. Radioactive parts security storage area, heat removal storage casks. ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

A&M. Radioactive parts security storage area, heat removal storage casks. Plan, section, and details. Ralph M. Parsons 1480-7 ANP/GE-3-720-S-1. Date: November 1958. Approved by INEEL Classification Office for public release. INEEL index no. 034-0720-60-693-107459 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID

78 FR 73379 - List of Approved Spent Fuel Storage Casks: HI-STORM 100 Cask System; Amendment No. 9

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-06

..., and criticality control. If there is no loss of confinement, shielding, or criticality control, the... would prevent loss of confinement, shielding, and criticality control. If there is no loss of...;Federal Register / Vol. 78, No. 235 / Friday, December 6, 2013 / Rules and Regulations#0;#0; [[Page 73379...

75 FR 42292 - List of Approved Spent Fuel Storage Casks: NAC-MPC System, Revision 6

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-21

... modifications to the Vertical Concrete Cask (VCC) incorporating design features from the MAGNASTOR system for...; an increase in the concrete pad compression strength from 4,000 psi to 6,000 psi; added justification... system while adhering to ALARA principles; (5) an increase in the concrete pad compression strength from...

Separator assembly for use in spent nuclear fuel shipping cask

DOEpatents

Bucholz, James A.

1983-01-01

A separator assembly for use in a spent nuclear fuel shipping cask has a honeycomb-type wall structure defining parallel cavities for holding nuclear fuel assemblies. Tubes formed of an effective neutron-absorbing material are embedded in the wall structure around each of the cavities and provide neutron flux traps when filled with water.

Sorting nexin-2 is associated with tubular elements of the early endosome, but is not essential for retromer-mediated endosome-to-TGN transport

PubMed Central

Carlton, Jez G.; Bujny, Miriam V.; Peter, Brian J.; Oorschot, Viola M. J.; Rutherford, Anna; Arkell, Rebecca S.; Klumperman, Judith; McMahon, Harvey T.; Cullen, Peter J.

2006-01-01

Summary Sorting nexins are a large family of phox-homology-domain-containing proteins that have been implicated in the control of endosomal sorting. Sorting nexin-1 is a component of the mammalian retromer complex that regulates retrieval of the cation-independent mannose 6-phosphate receptor from endosomes to the trans-Golgi network. In yeast, retromer is composed of Vps5p (the orthologue of sorting nexin-1), Vps17p (a related sorting nexin) and a cargo selective subcomplex composed of Vps26p, Vps29p and Vps35p. With the exception of Vps17p, mammalian orthologues of all yeast retromer components have been identified. For Vps17p, one potential mammalian orthologue is sorting nexin-2. Here we show that, like sorting nexin-1, sorting nexin-2 binds phosphatidylinositol 3-monophosphate and phosphatidylinositol 3,5-bisphosphate, and possesses a Bin/Amphiphysin/Rvs domain that can sense membrane curvature. However, in contrast to sorting nexin-1, sorting nexin-2 could not induce membrane tubulation in vitro or in vivo. Functionally, we show that endogenous sorting nexin-1 and sorting nexin-2 co-localise on high curvature tubular elements of the 3-phosphoinositide-enriched early endosome, and that suppression of sorting nexin-2 does not perturb the degradative sorting of receptors for epidermal growth factor or transferrin, nor the steady-state distribution of the cation-independent mannose 6-phosphate receptor. However, suppression of sorting nexin-2 results in a subtle alteration in the kinetics of cation-independent mannose 6-phosphate receptor retrieval. These data suggest that although sorting nexin-2 may be a component of the retromer complex, its presence is not essential for the regulation of endosome-to-trans Golgi network retrieval of the cation-independent mannose 6-phosphate receptor. PMID:16179610

Rab7 Associates with Early Endosomes to Mediate Sorting and Transport of Semliki Forest Virus to Late Endosomes

PubMed Central

Vonderheit, Andreas

2005-01-01

Semliki forest virus (SFV) is internalized by clathrin-mediated endocytosis, and transported via early endosomes to late endosomes and lysosomes. The intracellular pathway taken by individual fluorescently labeled SFV particles was followed using immunofluorescence in untransfected cells, and by video-enhanced, triple-color fluorescence microscopy in live cells transfected with GFP- and RFP-tagged Rab5, Rab7, Rab4, and Arf1. The viruses progressed from Rab5-positive early endosomes to a population of early endosomes (about 10% of total) that contained both Rab5 and Rab7. SFV were sequestered in the Rab7 domains, and they were sorted away from the early endosomes when these domains detached as separate transport carriers devoid of Rab5, Rab4, EEA1, Arf1, and transferrin. The process was independent of Arf1 and the acidic pH in early endosomes. Nocodazole treatment showed that the release of transport carriers was assisted by microtubules. Expression of constitutively inactive Rab7T22N resulted in accumulation of SFV in early endosomes. We concluded that Rab7 is recruited to early endosomes, where it forms distinct domains that mediate cargo sorting as well as the formation of late-endosome-targeted transport vesicles. PMID:15954801

Activation of epidermal growth factor receptor mediates receptor axon sorting and extension in the developing olfactory system of the moth Manduca sexta.

PubMed

Gibson, Nicholas J; Tolbert, Leslie P

2006-04-10

During development of the adult olfactory system of the moth Manduca sexta, olfactory receptor neurons extend axons from the olfactory epithelium in the antenna into the brain. As they arrive at the brain, interactions with centrally derived glial cells cause axons to sort and fasciculate with other axons destined to innervate the same glomeruli. Here we report studies indicating that activation of the epidermal growth factor receptor (EGFR) is involved in axon ingrowth and targeting. Blocking the EGFR kinase domain pharmacologically leads to stalling of many axons in the sorting zone and nerve layer as well as abnormal axonal fasciculation in the sorting zone. We also find that neuroglian, an IgCAM known to activate the EGFR through homophilic interactions in other systems, is transiently present on olfactory receptor neuron axons and on glia during the critical stages of the sorting process. The neuroglian is resistant to extraction with Triton X-100 in the sorting zone and nerve layer, possibly indicating its stabilization by homophilic binding in these regions. Our results suggest a mechanism whereby neuroglian molecules on axons and possibly sorting zone glia bind homophilically, leading to activation of EGFRs, with subsequent effects on axon sorting, pathfinding, and extension, and glomerulus development. Copyright 2006 Wiley-Liss, Inc.

Activation of EGF Receptor Mediates Receptor Axon Sorting and Extension in the Developing Olfactory System of the Moth Manduca sexta

PubMed Central

Gibson, Nicholas J.; Tolbert, Leslie P.

2008-01-01

During development of the adult olfactory system of the moth Manduca sexta, olfactory receptor neurons extend axons from the olfactory epithelium in the antenna into the brain. As they arrive at the brain, interactions with centrally-derived glial cells cause axons to sort and fasciculate with other axons destined to innervate the same glomeruli. Here we report studies that indicate that activation of the epidermal growth factor receptor (EGFR) is involved in axon ingrowth and targeting. Blocking the EGFR kinase domain pharmacologically leads to stalling of many axons in the sorting zone and nerve layer, as well as abnormal axonal fasciculation in the sorting zone. We also find that neuroglian, an IgCAM known to activate the EGFR through homophilic interactions in other systems, is transiently present on olfactory receptor neuron axons and on glia during the critical stages of the sorting process. The neuroglian is resistant to extraction with Triton X-100 in the sorting zone and nerve layer, possibly indicating its stabilization by homophilic binding in these regions. Our results suggest a mechanism whereby neuroglian molecules on axons and possibly sorting zone glia bind homophilically, leading to activation of EGFRs with subsequent effects on axon sorting, pathfinding, and extension, and glomerulus development. PMID:16498681

75 FR 57841 - List of Approved Spent Fuel Storage Casks: NAC-MPC System, Revision 6, Confirmation of Effective...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-23

... Spent Fuel Storage Casks: NAC-MPC System, Revision 6, Confirmation of Effective Date AGENCY: Nuclear... include Amendment Number 6 to Certificate of Compliance (CoC) Number 1025. DATES: Effective Date: The... regulations at 10 CFR 72.214 to include Amendment No. 6 to CoC No. 1025. Amendment No. 6 changes the...

78 FR 8050 - Spent Fuel Cask Certificate of Compliance Format and Content

Federal Register 2010, 2011, 2012, 2013, 2014

2013-02-05

... Rule for CoC Format and Content The petitioner states that amending 10 CFR part 72, subpart L, to... conforming changes be made to 10 CFR 72.13. The petitioner argues that ``[n]ew or amended NRC staff positions... 72, subpart L, be amended to remove the requirement that the empty weight be marked on storage casks...

78 FR 67348 - Invitation for Public Comment on Draft Test Plan for the High Burnup Dry Storage Cask Research...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-11-12

... will be executed will be added when Dominion Virginia Power, who is part of the Electric Power research... Electric Power Research Institute (EPRI) to document what is planned to be accomplished by the CDP. DOE is... Storage Cask Research and Development Project (CDP) AGENCY: Fuel Cycle Technologies, Office of Nuclear...

Characterization of the radiation environment for a large-area interim spent-nuclear-fuel storage facility

NASA Astrophysics Data System (ADS)

Fortkamp, Jonathan C.

Current needs in the nuclear industry and movements in the political arena indicate that authorization may soon be given for development of a federal interim storage facility for spent nuclear fuel. The initial stages of the design work have already begun within the Department of Energy and are being reviewed by the Nuclear Regulatory Commission. This dissertation addresses the radiation environment around an interim spent nuclear fuel storage facility. Specifically the dissertation characterizes the radiation dose rates around the facility based on a design basis source term, evaluates the changes in dose due to varying cask spacing configurations, and uses these results to define some applicable health physics principles for the storage facility. Results indicate that dose rates from the facility are due primarily from photons from the spent fuel and Co-60 activation in the fuel assemblies. In the modeled cask system, skyshine was a significant contribution to dose rates at distances from the cask array, but this contribution can be reduced with an alternate cask venting system. With the application of appropriate health physics principles, occupation doses can be easily maintained far below regulatory limits and maintained ALARA.

Corrosion of aluminum clad spent nuclear fuel in the 70 ton cask during transfer from L area to H-canyon

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mickalonis, J. I.

2015-08-31

Aluminum-clad spent nuclear fuel will be transported for processing in the 70-ton nuclear fuel element cask from L Basin to H-canyon. During transport these fuels would be expected to experience high temperature aqueous corrosion from the residual L Basin water that will be present in the cask. Cladding corrosion losses during transport were calculated for material test reactor (MTR) and high flux isotope reactors (HFIR) fuels using literature and site information on aqueous corrosion at a range of time/temperature conditions. Calculations of the cladding corrosion loss were based on Arrhenius relationships developed for aluminum alloys typical of cladding material withmore » the primary assumption that an adherent passive film does not form to retard the initial corrosion rate. For MTR fuels a cladding thickness loss of 33 % was found after 1 year in the cask with a maximum temperature of 263 °C. HFIR fuels showed a thickness loss of only 6% after 1 year at a maximum temperature of 180 °C. These losses are not expected to impact the overall confinement function of the aluminum cladding.« less

Corrosion of aluminum clad spent nuclear fuel in the 70 ton cask during transfer from L area to H-canyon

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mickalonis, J. I.

2015-08-01

Aluminum-clad spent nuclear fuel will be transported for processing in the 70-ton nuclear fuel element cask from L Basin to H-canyon. During transport these fuels would be expected to experience high temperature aqueous corrosion from the residual L Basin water that will be present in the cask. Cladding corrosion losses during transport were calculated for material test reactor (MTR) and high flux isotope reactors (HFIR) fuels using literature and site information on aqueous corrosion at a range of time/temperature conditions. Calculations of the cladding corrosion loss were based on Arrhenius relationships developed for aluminum alloys typical of cladding material withmore » the primary assumption that an adherent passive film does not form to retard the initial corrosion rate. For MTR fuels a cladding thickness loss of 33% was found after 1 year in the cask with a maximum temperature of 263 °C. HFIR fuels showed a thickness loss of only 6% after 1 year at a maximum temperature of 180 °C. These losses are not expected to impact the overall confinement function of the aluminum cladding.« less

Safety evaluation for packaging (onsite) plutonium recycle test reactor graphite cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

Romano, T.

This safety evaluation for packaging (SEP) provides the evaluation necessary to demonstrate that the Plutonium Recycle Test Reactor (PRTR) Graphite Cask meets the requirements of WHC-CM-2-14, Hazardous Material Packaging and Shipping, for transfer of Type B, fissile, non-highway route controlled quantities of radioactive material within the 300 Area of the Hanford Site. The scope of this SEP includes risk, shieldling, criticality, and.tiedown analyses to demonstrate that onsite transportation safety requirements are satisfied. This SEP also establishes operational and maintenance guidelines to ensure that transport of the PRTR Graphite Cask is performed safely in accordance with WHC-CM-2-14. This SEP is validmore » until October 1, 1999. After this date, an update or upgrade to this document is required.« less

Storage, transportation and disposal system for used nuclear fuel assemblies

DOEpatents

Scaglione, John M.; Wagner, John C.

2017-01-10

An integrated storage, transportation and disposal system for used fuel assemblies is provided. The system includes a plurality of sealed canisters and a cask sized to receive the sealed canisters in side by side relationship. The plurality of sealed canisters include an internal basket structure to receive a plurality of used fuel assemblies. The internal basket structure includes a plurality of radiation-absorbing panels and a plurality of hemispherical ribs generally perpendicular to the canister sidewall. The sealed canisters are received within the cask for storage and transportation and are removed from the cask for disposal at a designated repository. The system of the present invention allows the handling of sealed canisters separately or collectively, while allowing storage and transportation of high burnup fuel and damaged fuel to the designated repository.

An Investigation into the Transportation of Irradiated Uranium/Aluminum Targets from a Foreign Nuclear Reactor to the Chalk River Laboratories Site in Ontario, Canada - 12249

DOE Office of Scientific and Technical Information (OSTI.GOV)

Clough, Malcolm; Jackson, Austin

2012-07-01

This investigation required the selection of a suitable cask and development of a device to hold and transport irradiated targets from a foreign nuclear reactor to the Chalk River Laboratories in Ontario, Canada. The main challenge was to design and validate a target holder to protect the irradiated HEU-Al target pencils during transit. Each of the targets was estimated to have an initial decay heat of 118 W prior to transit. As the targets have little thermal mass the potential for high temperature damage and possibly melting was high. Thus, the primary design objective was to conceive a target holdermore » to dissipate heat from the targets. Other design requirements included securing the targets during transportation and providing a simple means to load and unload the targets while submerged five metres under water. A unique target holder (patent pending) was designed and manufactured together with special purpose experimental apparatus including a representative cask. Aluminum dummy targets were fabricated to accept cartridge heaters, to simulate decay heat. Thermocouples were used to measure the temperature of the test targets and selected areas within the target holder and test cask. After obtaining test results, calculations were performed to compensate for differences between experimental and real life conditions. Taking compensation into consideration the maximum target temperature reached was 231 deg. C which was below the designated maximum of 250 deg. C. The design of the aluminum target holder also allowed generous clearance to insert and unload the targets. This clearance was designed to close up as the target holder is placed into the cavity of the transport cask. Springs served to retain and restrain the targets from movement during transportation as well as to facilitate conductive heat transfer. The target holder met the design requirements and as such provided data supporting the feasibility of transporting targets over a relatively long period of time. A suitable transport cask was selected and a device for housing irradiated targets for loading, unloading and transportation has been designed, built and validated. The device was successful in meeting all design requirements for this feasibility study. Experiments were conducted with a custom test facility to confirm that the design met the maximum temperature requirements during shipping. Results from tests showed that the peak temperature in the apparatus was 300 deg. C. By compensating for experimental considerations, such as reduced thermal conductivity of the test cask versus that of the actual cask the expected maximum target temperature reduces to 231 deg. C. This is below the designated peak value of 250 deg. C. It can therefore be concluded, based on the content of this paper and from a heat-removal standpoint, the feasibility of transporting targets from a foreign nuclear reactor to Canada is possible, although further testing with irradiated targets and a full size cask would be a recommended next step. (authors)« less

The yeast Alix homolog, Bro1, functions as a ubiquitin receptor for protein sorting into multivesicular endosomes

PubMed Central

Pashkova, Natasha; Gakhar, Lokesh; Winistorfer, Stanley; Sunshine, Anna B.; Rich, Matthew; Dunham, Maitreya J.; Yu, Liping; Piper, Robert

2013-01-01

SUMMARY Sorting of ubiquitinated membrane proteins into lumenal vesicles of multivesicular bodies is mediated by the ESCRT apparatus and accessory proteins such as Bro1, which recruits the deubiquitinating enzyme Doa4 to remove ubiquitin from cargo. Here we propose that Bro1 works as a receptor for the selective sorting of ubiquitinated cargos. We found synthetic genetic interactions between BRO1 and ESCRT-0, suggesting Bro1 functions similarly to ESCRT-0. Multiple structural approaches demonstrated that Bro1 binds ubiquitin via the N-terminal trihelical arm of its middle V domain. Mutants of Bro1 that lack the ability to bind Ub were dramatically impaired in their ability to sort Ub-cargo membrane proteins, but only when combined with hypomorphic alleles of ESCRT-0. These data suggest that Bro1 and other Bro1 family members function in parallel with ESCRT-0 to recognize and sort Ub-cargos. PMID:23726974

Improvement of operational safety of dual-purpose transport packaging set for naval SNF in storage

DOE Office of Scientific and Technical Information (OSTI.GOV)

Guskov, Vladimir; Korotkov, Gennady; Barnes, Ella

2007-07-01

Available in abstract form only. Full text of publication follows: In recent ten years a new technology of management of irradiated nuclear fuel (SNF) at the final stage of fuel cycle has been intensely developing on a basis of a new type of casks used for interim storage of SNF and subsequent transportation therein to the place of processing, further storage or final disposal. This technology stems from the concept of a protective cask which provides preservation of its content (SNF) and fulfillment of all other safety requirements for storage and transportation of SNF. Radiation protection against emissions and non-distributionmore » of activity outside the cask is ensured by physical barriers, i.e. all-metal or composite body, shells, inner cavities for irradiated fuel assemblies (SFA), lids with sealing systems. Residual heat release of SFA is discharged to the environment by natural way: through emission and convection of surrounding air. By now more than 100 dual purpose packaging sets TUK-108/1 are in operation in the mode of interim storage and transportation of SNF from decommissioned nuclear powered submarines (NPS). In accordance with certificate, spent fuel is stored in TUK-108/1 on the premises of plants involved in NPS dismantlement for 2 years, whereupon it is transported for processing to PO Mayak. At one Far Eastern plant Zvezda involved in NPS dismantlement there arose a complicated situation due to necessity to extend period of storage of SNF in TUK- 108/1. To ensure safety over a longer period of storage of SNF in TUK-108/1 it is essential to modify conditions of storage by removing of residual water and filling the inner cavity of the cask with an inert gas. Within implementation of the international 1.1- 2 project Development of drying technology for the cask TUK-108/1 intended for naval SNF under the Program, there has been developed the technology of preparation of the cask for long-term storage of SNF in TUK-108/1, the design of a mobile TUK-108/1 drying facility; a pilot facility has been manufactured. This report describes key issues of cask drying technology, justification of terms of dry storage of naval SNF in no-108/1, design features of the mobile drying facility, results of tests of the pilot facility at the Far Eastern plant Zvezda. (authors)« less

Spent nuclear fuel dry transfer system

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stewart, L.; Agace, S.

The U.S. Department of Energy is currently engaged in a cooperative program with the Electric Power Research Institute (EPRI) to design a spent nuclear fuel dry transfer system (DTS). The system will enable the transfer of individual spent nuclear fuel assemblies between a conventional top loading cask and multi-purpose canister in a shielded overpack, or accommodate spent nuclear fuel transfers between two conventional casks.

Safety evaluation for packaging for the transport of K Basin sludge samples in the PAS-1 cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

SMITH, R.J.

1998-11-17

This safety evaluation for packaging authorizes the shipment of up to two 4-L sludge samples to and from the 325 Lab or 222-S Lab for characterization. The safety of this shipment is based on the current U.S. Department of Energy Certification of Compliance (CoC) for the PAS-1 cask, USA/9184/B(U) (DOE).

Clathrin-mediated endocytosis is a candidate entry sorting mechanism for Bombyx mori cypovirus.

PubMed

Chen, Fei; Zhu, Liyuan; Zhang, Yiling; Kumar, Dhiraj; Cao, Guangli; Hu, Xiaolong; Liang, Zi; Kuang, Sulan; Xue, Renyu; Gong, Chengliang

2018-05-08

Bombyx mori cypovirus (BmCPV), a member of the Reoviridae, specifically infects silkworms and causes extensive economic losses to the sericulture industry. To date, the entry mechanism of BmCPV into cells is unclear. Here we used electron microscopy to study the route of entry of BmCPV into cells, and the results demonstrated that the entry of BmCPV into BmN cells was mediated by endocytosis. Blocking the entry pathway with four endocytosis inhibitors, including dansylcadaverine, chlorpromazine, genistein, and PP2, significantly decreased the infectivity of BmCPV. This indicates that BmCPV enters BmN cells via endocytosis, and that clathrin-mediated sorting is the predominant entry method. After the relative expression levels of clathrin heavy chain (clathrin, GenBank accession No. NM_001142971.1) and the adaptor protein complex-1 gamma subunit AP-1 (AP-1, GenBank accession No. JQ824201.1), which are involved in clathrin-mediated endocytosis, were inhibited by RNA interference or abolishing the functions of clathrin and AP-1 with their corresponding antibodies, the infectivity of BmCPV was reduced significantly, which suggests that clathrin-mediated endocytosis contributed to the entry of BmCPV into cells. Our findings suggest that the clathrin-mediated endocytosis pathway is a candidate for the development of therapeutics for silkworm cytoplasmic polyhedrosis.

Nonlinear Ultrasonic Diagnosis and Prognosis of ASR Damage in Dry Cask Storage

DOE Office of Scientific and Technical Information (OSTI.GOV)

Qu, Jianmin; Bazant, Zdenek; Jacobs, Laurence

Alkali-silica reaction (ASR) is a deleterious chemical process that may occur in cement-based materials such as mortars and concretes, where the hydroxyl ions in the highly alkaline pore solution attack the siloxane groups in the siliceous minerals in the aggregates. The reaction produces a cross-linked alkali-silica gel. The ASR gel swells in the presence of water. Expansion of the gel results in cracking when the swelling-induced stress exceeds the fracture toughness of the concrete. As the ASR continues, cracks may grow and eventually coalesce, which results in reduced service life and a decrease safety of concrete structures. Since concrete ismore » widely used as a critical structural component in dry cask storage of used nuclear fuels, ASR damage poses a significant threat to the sustainability of long term dry cask storage systems. Therefore, techniques for effectively detecting, managing and mitigating ASR damage are needed. Currently, there are no nondestructive methods to accurately detect ASR damage in existing concrete structures. The only current way of accurately assessing ASR damage is to drill a core from an existing structure, and conduct microscopy on this drilled cylindrical core. Clearly, such a practice is not applicable to dry cask storage systems. To meet these needs, this research is aimed at developing (1) a suite of nonlinear ultrasonic quantitative nondestructive evaluation (QNDE) techniques to characterize ASR damage, and (2) a physics-based model for ASR damage evolution using the QNDE data. Outcomes of this research will provide a nondestructive diagnostic tool to evaluate the extent of the ASR damage, and a prognostic tool to estimate the future reliability and safety of the concrete structures in dry cask storage systems« less

The Role of the Clathrin Adaptor AP-1: Polarized Sorting and Beyond

PubMed Central

Nakatsu, Fubito; Hase, Koji; Ohno, Hiroshi

2014-01-01

The selective transport of proteins or lipids by vesicular transport is a fundamental process supporting cellular physiology. The budding process involves cargo sorting and vesicle formation at the donor membrane and constitutes an important process in vesicular transport. This process is particularly important for the polarized sorting in epithelial cells, in which the cargo molecules need to be selectively sorted and transported to two distinct destinations, the apical or basolateral plasma membrane. Adaptor protein (AP)-1, a member of the AP complex family, which includes the ubiquitously expressed AP-1A and the epithelium-specific AP-1B, regulates polarized sorting at the trans-Golgi network and/or at the recycling endosomes. A growing body of evidence, especially from studies using model organisms and animals, demonstrates that the AP-1-mediated polarized sorting supports the development and physiology of multi-cellular units as functional organs and tissues (e.g., cell fate determination, inflammation and gut immune homeostasis). Furthermore, a possible involvement of AP-1B in the pathogenesis of human diseases, such as Crohn’s disease and cancer, is now becoming evident. These data highlight the significant contribution of AP-1 complexes to the physiology of multicellular organisms, as master regulators of polarized sorting in epithelial cells. PMID:25387275

Storage, transportation and disposal system for used nuclear fuel assemblies

DOE Office of Scientific and Technical Information (OSTI.GOV)

Scaglione, John M.; Wagner, John C.

An integrated storage, transportation and disposal system for used fuel assemblies is provided. The system includes a plurality of sealed canisters and a cask sized to receive the sealed canisters in side by side relationship. The plurality of sealed canisters include an internal basket structure to receive a plurality of used fuel assemblies. The internal basket structure includes a plurality of radiation-absorbing panels and a plurality of hemispherical ribs generally perpendicular to the canister sidewall. The sealed canisters are received within the cask for storage and transportation and are removed from the cask for disposal at a designated repository. Themore » system of the present invention allows the handling of sealed canisters separately or collectively, while allowing storage and transportation of high burnup fuel and damaged fuel to the designated repository.« less

ESCRT-III-Associated Protein ALIX Mediates High-Affinity Phosphate Transporter Trafficking to Maintain Phosphate Homeostasis in Arabidopsis

PubMed Central

Cardona-López, Ximena; Cuyas, Laura; Marín, Elena; Irigoyen, María Luisa; Gil, Erica; Puga, María Isabel; Bligny, Richard; Nussaume, Laurent; Geldner, Niko; Paz-Ares, Javier

2015-01-01

Prior to the release of their cargoes into the vacuolar lumen, sorting endosomes mature into multivesicular bodies (MVBs) through the action of ENDOSOMAL COMPLEX REQUIRED FOR TRANSPORT (ESCRT) protein complexes. MVB-mediated sorting of high-affinity phosphate transporters (PHT1) to the vacuole limits their plasma membrane levels under phosphate-sufficient conditions, a process that allows plants to maintain phosphate homeostasis. Here, we describe ALIX, a cytosolic protein that associates with MVB by interacting with ESCRT-III subunit SNF7 and mediates PHT1;1 trafficking to the vacuole in Arabidopsis thaliana. We show that the partial loss-of-function mutant alix-1 displays reduced vacuolar degradation of PHT1;1. ALIX derivatives containing the alix-1 mutation showed reduced interaction with SNF7, providing a simple molecular explanation for impaired cargo trafficking in alix-1 mutants. In fact, the alix-1 mutation also hampered vacuolar sorting of the brassinosteroid receptor BRI1. We also show that alix-1 displays altered vacuole morphogenesis, implying a new role for ALIX proteins in vacuolar biogenesis, likely acting as part of ESCRT-III complexes. In line with a presumed broad target spectrum, the alix-1 mutation is pleiotropic, leading to reduced plant growth and late flowering, with stronger alix mutations being lethal, indicating that ALIX participates in diverse processes in plants essential for their life. PMID:26342016

National Policy Implications of Storing Nuclear Waste in the Pacific Region,

DTIC Science & Technology

1981-01-01

US Congress, Senate, Committee on Energy and Natural Resources, Pacific Spent Nuclear Fuel Storage , Hearing...selected. 17 One type of shipping cask which has been used to transport spent fuel assemblies to the Nevada Test Site is a leakproof steel cask that can...discussion the following conclusions on the nuclear waste storage issue appear valid. The Reagan decision to reprocess spent fuel has not changed US

Performance of bolted closure joint elastomers under cask aging conditions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Verst, C.; Sindelar, R.; Skidmore, E.

The bolted closure joint of a bare spent fuel cask is susceptible to age-related degradation and potential loss of confinement function under long-term storage conditions. Elastomeric seals, a component of the joint typically used to facilitate leak testing of the primary seal that includes the metallic seal and bolting, is susceptible to degradation over time by several mechanisms, principally via thermo-oxidation, stress-relaxation, and radiolytic degradation under time and temperature condition. Irradiation and thermal exposure testing and evaluation of an ethylene-propylene diene monomer (EPDM) elastomeric seal material similar to that used in the CASTOR® V/21 cask for a matrix of temperaturemore » and radiation exposure conditions relevant to the cask extended storage conditions, and development of semiempirical predictive models for loss of sealing force is in progress. A special insert was developed to allow Compressive Stress Relaxation (CSR) measurements before and after the irradiation and/or thermal exposure without unloading the elastomer. A condition of the loss of sealing force for the onset of leakage was suggested. The experimentation and modeling being performed could enable acquisition of extensive coupled aging data as well as an estimation of the timeframe when loss of sealing function under aging (temperature/radiation) conditions may occur.« less

Alternative Splicing of a Novel Inducible Exon Diversifies the CASK Guanylate Kinase Domain

PubMed Central

Dembowski, Jill A.; An, Ping; Scoulos-Hanson, Maritsa; Yeo, Gene; Han, Joonhee; Fu, Xiang-Dong; Grabowski, Paula J.

2012-01-01

Alternative pre-mRNA splicing has a major impact on cellular functions and development with the potential to fine-tune cellular localization, posttranslational modification, interaction properties, and expression levels of cognate proteins. The plasticity of regulation sets the stage for cells to adjust the relative levels of spliced mRNA isoforms in response to stress or stimulation. As part of an exon profiling analysis of mouse cortical neurons stimulated with high KCl to induce membrane depolarization, we detected a previously unrecognized exon (E24a) of the CASK gene, which encodes for a conserved peptide insertion in the guanylate kinase interaction domain. Comparative sequence analysis shows that E24a appeared selectively in mammalian CASK genes as part of a >3,000 base pair intron insertion. We demonstrate that a combination of a naturally defective 5′ splice site and negative regulation by several splicing factors, including SC35 (SRSF2) and ASF/SF2 (SRSF1), drives E24a skipping in most cell types. However, this negative regulation is countered with an observed increase in E24a inclusion after neuronal stimulation and NMDA receptor signaling. Taken together, E24a is typically a skipped exon, which awakens during neuronal stimulation with the potential to diversify the protein interaction properties of the CASK polypeptide. PMID:23008758

Distinct forms of mitochondrial TOM-TIM supercomplexes define signal-dependent states of preprotein sorting.

PubMed

Chacinska, Agnieszka; van der Laan, Martin; Mehnert, Carola S; Guiard, Bernard; Mick, David U; Hutu, Dana P; Truscott, Kaye N; Wiedemann, Nils; Meisinger, Chris; Pfanner, Nikolaus; Rehling, Peter

2010-01-01

Mitochondrial import of cleavable preproteins occurs at translocation contact sites, where the translocase of the outer membrane (TOM) associates with the presequence translocase of the inner membrane (TIM23) in a supercomplex. Different views exist on the mechanism of how TIM23 mediates preprotein sorting to either the matrix or inner membrane. On the one hand, two TIM23 forms were proposed, a matrix transport form containing the presequence translocase-associated motor (PAM; TIM23-PAM) and a sorting form containing Tim21 (TIM23(SORT)). On the other hand, it was reported that TIM23 and PAM are permanently associated in a single-entity translocase. We have accumulated distinct transport intermediates of preproteins to analyze the translocases in their active, preprotein-carrying state. We identified two different forms of active TOM-TIM23 supercomplexes, TOM-TIM23(SORT) and TOM-TIM23-PAM. These two supercomplexes do not represent separate pathways but are in dynamic exchange during preprotein translocation and sorting. Depending on the signals of the preproteins, switches between the different forms of supercomplex and TIM23 are required for the completion of preprotein import.

Sortilin 1 Loss-of-Function Protects Against Cholestatic Liver Injury by Attenuating Hepatic Bile Acid Accumulation in Bile Duct Ligated Mice.

PubMed

Li, Jibiao; Woolbright, Benjamin L; Zhao, Wen; Wang, Yifeng; Matye, David; Hagenbuch, Bruno; Jaeschke, Hartmut; Li, Tiangang

2018-01-01

Sortilin 1 (Sort1) is an intracellular trafficking receptor that mediates protein sorting in the endocytic or secretory pathways. Recent studies revealed a role of Sort1 in the regulation of cholesterol and bile acid (BA) metabolism. This study further investigated the role of Sort1 in modulating BA detoxification and cholestatic liver injury in bile duct ligated mice. We found that Sort1 knockout (KO) mice had attenuated liver injury 24 h after bile duct ligation (BDL), which was mainly attributed to less bile infarct formation. Sham-operated Sort1 KO mice had about 20% larger BA pool size than sham-operated wildtype (WT) mice, but 24 h after BDL Sort1 KO mice had significantly attenuated hepatic BA accumulation and smaller BA pool size. After 14 days BDL, Sort1 KO mice showed significantly lower hepatic BA concentration and reduced expression of inflammatory and fibrotic marker genes, but similar degree of liver fibrosis compared with WT mice. Unbiased quantitative proteomics revealed that Sort1 KO mice had increased hepatic BA sulfotransferase 2A1, but unaltered phase-I BA metabolizing cytochrome P450s or phase-III BA efflux transporters. Consistently, Sort1 KO mice showed elevated plasma sulfated taurocholate after BDL. Finally, we found that liver Sort1 was repressed after BDL, which may be due to BA activation of farnesoid x receptor. In conclusion, we report a role of Sort1 in the regulation of hepatic BA detoxification and cholestatic liver injury in mice. The mechanisms underlying increased hepatic BA elimination in Sort1 KO mice after BDL require further investigation. © The Author 2017. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Discovery of Prostate Cancer Tumor Suppressors and Mediators of MDV3100 Resistance through in Vivo RNA Interference Screen

DTIC Science & Technology

2014-05-01

NE phenotype. Figure 5: Rational of the FACS-based screen. Left, the concept of increase of NSE and decrease of PSMA with NED. Right, the...of AR dependency is associated with increase in Prostate specific membrane antigen ( PSMA ) expression. Thus, we also decided to sort for cells that...have decreased PSMA expression with prolonged Enzalutamide treatment. Using the above markers and FACS we attempted to sort out 4 populations of

Neutron flux measurements on a mock-up of a storage cask for high-level nuclear waste using 2.5 MeV neutrons.

PubMed

Suárez, H Saurí; Becker, F; Klix, A; Pang, B; Döring, T

2018-06-07

To store and dispose spent nuclear fuel, shielding casks are employed to reduce the emitted radiation. To evaluate the exposure of employees handling such casks, Monte Carlo radiation transport codes can be employed. Nevertheless, to assess the reliability of these codes and nuclear data, experimental checks are required. In this study, a neutron generator (NG) producing neutrons of 2.5 MeV was employed to simulate neutrons produced in spent nuclear fuel. Different configurations of shielding layers of steel and polyethylene were positioned between the target of the NG and a NE-213 detector. The results of the measurements of neutron and γ radiation and the corresponding simulations with the code MCNP6 are presented. Details of the experimental set-up as well as neutron and photon flux spectra are provided as reference points for such NG investigations with shielding structures.

Fuel shipment experience, fuel movements from the BMI-1 transport cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bauer, Thomas L.; Krause, Michael G

1986-07-01

The University of Texas at Austin received two shipments of irradiated fuel elements from Northrup Aircraft Corporation on April 11 and 16, 1985. A total of 59 elements consisting of standard and instrumented TRIGA fuel were unloaded from the BMI-1 shipping cask. At the time of shipment, the Northrup core burnup was approximately 50 megawatt days with fuel element radiation levels, after a cooling time of three months, of approximately 1.75 rem/hr at 3 feet. In order to facilitate future planning of fuel shipment at the UT facility and other facilities, a summary of the recent transfer process including severalmore » factors which contributed to its success are presented. Numerous color slides were made of the process for future reference by UT and others involved in fuel transfer and handling of the BMI-1 cask.« less

Congeners in sugar cane spirits aged in casks of different woods.

PubMed

Bortoletto, Aline M; Alcarde, André R

2013-08-15

The profile of volatile compounds and aging markers in sugar cane spirits aged for 36 months in casks made of 10 types of wood were studied. The ethanol content, volatile acidity, aldehydes, esters, higher alcohols, and methanol were determined. In addition, gallic, vanilic and syringic acids, siringaldehyde, coniferaldehyde, sinapaldehyde, vanillin, 5-hydroxymethylfurfural and furfural were identified and quantified. The profile of volatile compounds characterised aging in each type of wood. The beverage aged in oak cask achieved the highest contents of maturation-related congeners. The Brazilian woods, similar to oak, were jequitibá rosa and cerejeira, which presented the highest contents of some maturation-related compounds, such as vanillin, vanilic acid, syringaldehyde and sinapaldehyde. Although oak wood conferred more chemical complexity to the beverage, Brazilian woods, singly or complementarily, present potential for spirit characterisation and for improving the quality of sugar cane spirits. Copyright © 2013 Elsevier Ltd. All rights reserved.

Characterization of Hydrogen Embrittled Zircaloy-4 by Using a Van de Graaff Particle Accelerator

NASA Astrophysics Data System (ADS)

Budd, John

2013-04-01

On-site, dry cask storage was originally by the intended to be a short-term solution for holding spent nuclear fuel. Due to the lack of a permanent storage facility, the nuclear power industry seeks to assess the effective lifetime of the casks. One issue which could compromise cask integrity is Hydrogen embrittlement. This phenomenon occurs in the Zircaloy-4 fuel-rod cladding and is caused by the formation of Zirconium hydrides. Over time, thermal stresses caused by the heat from reactions of the stored nuclear fuel could result in significant breaches of the cladding. Our group at Texas A&M University- Kingsville is conducting experiments to aid in determining when such breaches will occur. We will irradiate samples of the alloy with protons of energies up to 400 keV using a Van de Graaff particle accelerator. Once irradiated, their properties will be characterized using scanning electron microscopy and Vickers hardness tests.

Secretory cargo sorting by Ca2+-dependent Cab45 oligomerization at the trans-Golgi network

PubMed Central

Blank, Birgit; Maiser, Andreas; Emin, Derya; Prescher, Jens; Beck, Gisela; Kienzle, Christine; Bartnik, Kira; Habermann, Bianca; Pakdel, Mehrshad; Leonhardt, Heinrich; Lamb, Don C.

2016-01-01

Sorting and export of transmembrane cargoes and lysosomal hydrolases at the trans-Golgi network (TGN) are well understood. However, elucidation of the mechanism by which secretory cargoes are segregated for their release into the extracellular space remains a challenge. We have previously demonstrated that, in a reaction that requires Ca2+, the soluble TGN-resident protein Cab45 is necessary for the sorting of secretory cargoes at the TGN. Here, we report that Cab45 reversibly assembles into oligomers in the presence of Ca2+. These Cab45 oligomers specifically bind secretory proteins, such as COMP and LyzC, in a Ca2+-dependent manner in vitro. In intact cells, mutation of the Ca2+-binding sites in Cab45 impairs oligomerization, as well as COMP and LyzC sorting. Superresolution microscopy revealed that Cab45 colocalizes with secretory proteins and the TGN Ca2+ pump (SPCA1) in specific TGN microdomains. These findings reveal that Ca2+-dependent changes in Cab45 mediate sorting of specific cargo molecules at the TGN. PMID:27138253

The endocytic recycling compartment maintains cargo segregation acquired upon exit from the sorting endosome

PubMed Central

Xie, Shuwei; Bahl, Kriti; Reinecke, James B.; Hammond, Gerald R. V.; Naslavsky, Naava; Caplan, Steve

2016-01-01

The endocytic recycling compartment (ERC) is a series of perinuclear tubular and vesicular membranes that regulates recycling to the plasma membrane. Despite evidence that cargo is sorted at the early/sorting endosome (SE), whether cargo mixes downstream at the ERC or remains segregated is an unanswered question. Here we use three-dimensional (3D) structured illumination microscopy and dual-channel and 3D direct stochastic optical reconstruction microscopy (dSTORM) to obtain new information about ERC morphology and cargo segregation. We show that cargo internalized either via clathrin-mediated endocytosis (CME) or independently of clathrin (CIE) remains segregated in the ERC, likely on distinct carriers. This suggests that no further sorting occurs upon cargo exit from SE. Moreover, 3D dSTORM data support a model in which some but not all ERC vesicles are tethered by contiguous “membrane bridges.” Furthermore, tubular recycling endosomes preferentially traffic CIE cargo and may originate from SE membranes. These findings support a significantly altered model for endocytic recycling in mammalian cells in which sorting occurs in peripheral endosomes and segregation is maintained at the ERC. PMID:26510502

Learning Style and Task Performance in Synchronous Computer-Mediated Communication: A Case Study of Iranian EFL Learners

ERIC Educational Resources Information Center

Hedayati, Mohsen; Foomani, Elham Mohammadi

2015-01-01

The study reported here explores whether English as a foreign Language (EFL) learners' preferred ways of learning (i.e., learning styles) affect their task performance in computer-mediated communication (CMC). As Ellis (2010) points out, while the increasing use of different sorts of technology is witnessed in language learning contexts, it is…

Children's Use of Semantic Organizational Strategies Is Mediated by Working Memory Capacity

ERIC Educational Resources Information Center

Schleepen, Tamara M. J.; Jonkman, Lisa M.

2012-01-01

In adults, the ability to apply semantic grouping strategies has been found to depend on working memory. To investigate this relation in children, two sort-recall tasks (one without and one with a grouping instruction) were administered to 6-12-year-olds. The role of working memory was examined by means of mediation analyses and by assessing…

The Lexical Stroop Sort (LSS) picture-word task: a computerized task for assessing the relationship between language and executive functioning in school-aged children.

PubMed

Wilbourn, Makeba Parramore; Kurtz, Laura E; Kalia, Vrinda

2012-03-01

The relationship between language development and executive function (EF) in children is not well understood. The Lexical Stroop Sort (LSS) task is a computerized EF task created for the purpose of examining the relationship between school-aged children's oral language development and EF. To validate this new measure, a diverse sample of school-aged children completed standardized oral language assessments, the LSS task, and the widely used Dimensional Change Card Sort (DCCS; Zelazo, 2006) task. Both EF tasks require children to sort stimuli into categories based on predetermined rules. While the DCCS largely relies on visual stimuli, the LSS employs children's phonological loop to access their semantic knowledge base. Accuracy and reaction times were recorded for both tasks. Children's scores on the LSS task were correlated with their scores on the DCCS task, and a similar pattern of relationships emerged between children's vocabulary and the two EF tasks, thus providing convergent validity for the LSS. However, children's phonological awareness was associated with their scores on the LSS, but not with those on the DCCS. In addition, a mediation model was used to elucidate the predictive relationship between phonological awareness and children's performance on the LSS task, with children's vocabulary fully mediating this relationship. The use of this newly created and validated LSS task with different populations, such as preschoolers and bilinguals, is also discussed.

The overexpressed human 46-kDa mannose 6-phosphate receptor mediates endocytosis and sorting of. beta. -glucuronidase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Watanabe, H.; Grubb, J.H.; Sly, W.S.

1990-10-01

The authors studied the function of the human small (46-kDa) mannose 6-phosphate receptor (SMPR) in transfected mouse L cells that do not express the larger insulin-like growth factor II/mannose 6-phosphate receptor. Cells overexpressing human SMPR were studied for enzyme binding to cell surface receptors, for binding to intracellular receptors in permeabilized cells, and for receptor-mediated endocytosis of recombinant human {beta}-glucuronidase. Specific binding to human SMPR in permeabilized cells showed a pH optimum between pH 6.0 and pH 6.5. Binding was significant in the present of EDTA but was enhanced by added divalent cations. Up to 2.3{percent} of the total functionalmore » receptor could be detected on the cell surface by enzyme binding. They present experiments showing that at very high levels of overexpression, and at pH 6.5, human SMPR mediated the endocytosis of {beta}-glucuronidase. At pH 7.5, the rate of endocytosis was only 14{percent} the rate seen at pH 6.5. Cells overexpressing human SMPR also showed reduced secretion of newly synthesized {beta}-glucuronidase when compared to cells transfected with vector only, suggesting that overexpressed human SMPR can participate in sorting of newly synthesized {beta}-glucuronidase and partially correct the sorting defect in mouse L cells that do not express the insulin-like growth factor II/mannose 6-phosphate receptor.« less

ALG-2 activates the MVB sorting function of ALIX through relieving its intramolecular interaction

PubMed Central

Sun, Sheng; Zhou, Xi; Corvera, Joe; Gallick, Gary E; Lin, Sue-Hwa; Kuang, Jian

2015-01-01

The modular adaptor protein ALIX is critically involved in endosomal sorting complexes required for transport (ESCRT)-mediated multivesicular body (MVB) sorting of activated epidermal growth factor receptor (EGFR); however, ALIX contains a default intramolecular interaction that renders ALIX unable to perform this ESCRT function. The ALIX partner protein ALG-2 is a calcium-binding protein that belongs to the calmodulin superfamily. Prompted by a defined biological function of calmodulin, we determined the role of ALG-2 in regulating ALIX involvement in MVB sorting of activated EGFR. Our results show that calcium-dependent ALG-2 interaction with ALIX completely relieves the intramolecular interaction of ALIX and promotes CHMP4-dependent ALIX association with the membrane. EGFR activation induces increased ALG-2 interaction with ALIX, and this increased interaction is responsible for increased ALIX association with the membrane. Functionally, inhibition of ALIX activation by ALG-2 inhibits MVB sorting of activated EGFR as effectively as inhibition of ALIX interaction with CHMP4 does; however, inhibition of ALIX activation by ALG-2 does not affect cytokinetic abscission or equine infectious anemia virus (EIAV) budding. These findings indicate that calcium-dependent ALG-2 interaction with ALIX is specifically responsible for generating functional ALIX that supports MVB sorting of ubiquitinated membrane receptors. PMID:27462417

LEVERAGING AGING MATERIALS DATA TO SUPPORT EXTENSION OF TRANSPORTATION SHIPPING PACKAGES SERVICE LIFE

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dunn, K.; Bellamy, S.; Daugherty, W.

Nuclear material inventories are increasingly being transferred to interim storage locations where they may reside for extended periods of time. Use of a shipping package to store nuclear materials after the transfer has become more common for a variety of reasons. Shipping packages are robust and have a qualified pedigree for performance in normal operation and accident conditions but are only certified over an approved transportation window. The continued use of shipping packages to contain nuclear material during interim storage will result in reduced overall costs and reduced exposure to workers. However, the shipping package materials of construction must maintainmore » integrity as specified by the safety basis of the storage facility throughout the storage period, which is typically well beyond the certified transportation window. In many ways, the certification processes required for interim storage of nuclear materials in shipping packages is similar to life extension programs required for dry cask storage systems for commercial nuclear fuels. The storage of spent nuclear fuel in dry cask storage systems is federally-regulated, and over 1500 individual dry casks have been in successful service up to 20 years in the US. The uncertainty in final disposition will likely require extended storage of this fuel well beyond initial license periods and perhaps multiple re-licenses may be needed. Thus, both the shipping packages and the dry cask storage systems require materials integrity assessments and assurance of continued satisfactory materials performance over times not considered in the original evaluation processes. Test programs for the shipping packages have been established to obtain aging data on materials of construction to demonstrate continued system integrity. The collective data may be coupled with similar data for the dry cask storage systems and used to support extending the service life of shipping packages in both transportation and storage.« less

Flow cytometric sex sorting affects CD4 membrane distribution and binding of exogenous DNA on bovine sperm cells.

PubMed

Domingues, William Borges; da Silveira, Tony Leandro Rezende; Komninou, Eliza Rossi; Monte, Leonardo Garcia; Remião, Mariana Härter; Dellagostin, Odir Antônio; Corcini, Carine Dahl; Varela Junior, Antônio Sergio; Seixas, Fabiana Kömmling; Collares, Tiago; Campos, Vinicius Farias

2017-08-01

Bovine sex-sorted sperm have been commercialized and successfully used for the production of transgenic embryos of the desired sex through the sperm-mediated gene transfer (SMGT) technique. However, sex-sorted sperm show a reduced ability to internalize exogenous DNA. The interaction between sperm cells and the exogenous DNA has been reported in other species to be a CD4-like molecule-dependent process. The flow cytometry-based sex-sorting process subjects the spermatozoa to different stresses causing changes in the cell membrane. The aim of this study was to elucidate the relationship between the redistribution of CD4-like molecules and binding of exogenous DNA to sex-sorted bovine sperm. In the first set of experiments, the membrane phospholipid disorder and the redistribution of the CD4 were evaluated. The second set of experiments was conducted to investigate the effect of CD4 redistribution on the mechanism of binding of exogenous DNA to sperm cells and the efficiency of lipofection in sex-sorted bovine sperm. Sex-sorting procedure increased the membrane phospholipid disorder and induced the redistribution of CD4-like molecules. Both X-sorted and Y-sorted sperm had decreased DNA bound to membrane in comparison with the unsorted sperm; however, the binding of the exogenous DNA was significantly increased with the addition of liposomes. Moreover, we demonstrated that the number of sperm-bound exogenous DNA was decreased when these cells were preincubated with anti-bovine CD4 monoclonal antibody, supporting our hypothesis that CD4-like molecules indeed play a crucial role in the process of exogenous DNA/bovine sperm cells interaction.

Final design review summary report for the TN-WHC cask and transportation system

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kee, A.T.

1997-01-17

This document represents comments generated from a review of Transnuclear`s Final Design Package distributed on December 10, 1996 and a review of the Final Design Analysis Report meeting held on December 17 & 18, 1996. The Final design describes desicn features and presents final analyses @j performed to fabricate and operate the system while meeting the Cask/Transportation Functions and Requirements, WHC-SD-SNF-FRD-011, Rev. 0 and specification WHC-S-0396, Rev. 1.

Safety analysis report for packaging (onsite) multicanister overpack cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

Edwards, W.S.

1997-07-14

This safety analysis report for packaging (SARP) documents the safety of shipments of irradiated fuel elements in the MUlticanister Overpack (MCO) and MCO Cask for a highway route controlled quantity, Type B fissile package. This SARP evaluates the package during transfers of (1) water-filled MCOs from the K Basins to the Cold Vacuum Drying Facility (CVDF) and (2) sealed and cold vacuum dried MCOs from the CVDF in the 100 K Area to the Canister Storage Building in the 200 East Area.

Snx3 regulates recycling of the transferrin receptor and iron assimilation

PubMed Central

Chen, Caiyong; Garcia-Santos, Daniel; Ishikawa, Yuichi; Seguin, Alexandra; Li, Liangtao; Fegan, Katherine H.; Hildick-Smith, Gordon J.; Shah, Dhvanit I.; Cooney, Jeffrey D.; Chen, Wen; King, Matthew J.; Yien, Yvette Y.; Schultz, Iman J.; Anderson, Heidi; Dalton, Arthur J.; Freedman, Matthew L.; Kingsley, Paul D.; Palis, James; Hattangadi, Shilpa M.; Lodish, Harvey F.; Ward, Diane M.; Kaplan, Jerry; Maeda, Takahiro; Ponka, Prem; Paw, Barry H.

2013-01-01

SUMMARY Sorting of endocytic ligands and receptors is critical for diverse cellular processes. The physiological significance of endosomal sorting proteins in vertebrates, however, remains largely unknown. Here we report that sorting nexin 3 (Snx3) facilitates the recycling of transferrin receptor (Tfrc), and thus is required for the proper delivery of iron to erythroid progenitors. Snx3 is highly expressed in vertebrate hematopoietic tissues. Silencing of Snx3 results in anemia and hemoglobin defects in vertebrates due to impaired transferrin (Tf)-mediated iron uptake and its accumulation in early endosomes. This impaired iron assimilation can be complemented with non-Tf iron chelates. We show that Snx3 and Vps35, a component of the retromer, interact with Tfrc to sort it to the recycling endosomes. Our findings uncover a role of Snx3 in regulating Tfrc recycling, iron homeostasis, and erythropoiesis. Thus, the identification of Snx3 provides a genetic tool for exploring erythropoiesis and disorders of iron metabolism. PMID:23416069

ARH directs megalin to the endocytic recycling compartment to regulate its proteolysis and gene expression

PubMed Central

Shah, Mehul; Baterina, Oscar Y.; Taupin, Vanessa

2013-01-01

Receptors internalized by endocytosis can return to the plasma membrane (PM) directly from early endosomes (EE; fast recycling) or they can traffic from EE to the endocytic recycling compartment (ERC) and recycle from there (slow recycling). How receptors are sorted for trafficking along these two pathways remains unclear. Here we show that autosomal recessive hypercholesterolemia (ARH) is required for trafficking of megalin, a member of the LDL receptor family, from EE to the ERC by coupling it to dynein; in the absence of ARH, megalin returns directly to the PM from EE via the connecdenn2/Rab35 fast recycling pathway. Binding of ARH to the endocytic adaptor AP-2 prevents fast recycling of megalin. ARH-mediated trafficking of megalin to the ERC is necessary for γ-secretase mediated cleavage of megalin and release of a tail fragment that mediates transcriptional repression. These results identify a novel mechanism for sorting receptors for trafficking to the ERC and link ERC trafficking to regulated intramembrane proteolysis (RIP) and expression of megalin. PMID:23836931

UNC93B1 mediates differential trafficking of endosomal TLRs

PubMed Central

Lee, Bettina L; Moon, Joanne E; Shu, Jeffrey H; Yuan, Lin; Newman, Zachary R; Schekman, Randy; Barton, Gregory M

2013-01-01

UNC93B1, a multipass transmembrane protein required for TLR3, TLR7, TLR9, TLR11, TLR12, and TLR13 function, controls trafficking of TLRs from the endoplasmic reticulum (ER) to endolysosomes. The mechanisms by which UNC93B1 mediates these regulatory effects remain unclear. Here, we demonstrate that UNC93B1 enters the secretory pathway and directly controls the packaging of TLRs into COPII vesicles that bud from the ER. Unlike other COPII loading factors, UNC93B1 remains associated with the TLRs through post-Golgi sorting steps. Unexpectedly, these steps are different among endosomal TLRs. TLR9 requires UNC93B1-mediated recruitment of adaptor protein complex 2 (AP-2) for delivery to endolysosomes while TLR7, TLR11, TLR12, and TLR13 utilize alternative trafficking pathways. Thus, our study describes a mechanism for differential sorting of endosomal TLRs by UNC93B1, which may explain the distinct roles played by these receptors in certain autoimmune diseases. DOI: http://dx.doi.org/10.7554/eLife.00291.001 PMID:23426999

Quantity and management of spent fuel from prototype and research reactors in Germany

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dorr, Sabine; Bollingerfehr, Wilhelm; Filbert, Wolfgang

Within the scope of an R and D project (project identification number FKZ 02 S 8679) sponsored by BMBF (Federal Ministry of Education and Research), the current state of storage and management of fuel elements from prototype and research reactors was established, and an approach for their future storage/management was developed. The spent fuels from prototype and research reactors in Germany that require disposal were specified and were described in regard to their repository-relevant characteristics. As there are currently no casks licensed for disposal in Germany, descriptions of casks that were considered to be suitable were provided. Based on themore » information provided on the spent fuel from prototype and research reactors and the potential casks, a technical disposal concept was developed. In this context, concepts to integrate the spent fuel from prototype and research reactors into existing disposal concepts for spent fuel from German nuclear power plants and for waste from reprocessing were developed for salt and clay formations. (authors)« less

Measurement of chlorine concentration on steel surfaces via fiber-optic laser-induced breakdown spectroscopy in double-pulse configuration

NASA Astrophysics Data System (ADS)

Xiao, X.; Le Berre, S.; Fobar, D. G.; Burger, M.; Skrodzki, P. J.; Hartig, K. C.; Motta, A. T.; Jovanovic, I.

2018-03-01

The corrosive environment provided by chlorine ions on the welds of stainless steel dry cask storage canisters for used nuclear fuel may contribute to the occurrence of stress corrosion cracking. We demonstrate the use of fiber-optic laser-induced breakdown spectroscopy (FOLIBS) in the double-pulse (DP) configuration for high-sensitivity, remote measurement of the surface concentrations of chlorine compatible in constrained space and challenging environment characteristic for dry cask storage systems. Chlorine surface concentrations as low as 5 mg/m2 have been detected and quantified by use of a laboratory-based and a fieldable DP FOLIBS setup with the calibration curve approach. The compact final optics assembly in the fieldable setup is interfaced via two 25-m long optical fibers for high-power laser pulse delivery and plasma emission collection and can be readily integrated into a multi-sensor robotic delivery system for in-situ inspection of dry cask storage systems.

Nuclear cask testing films misleading and misused

DOE Office of Scientific and Technical Information (OSTI.GOV)

Audin, L.

In 1977 and 1978, Sandia National Laboratories, located in Albuquerque, New Mexico, and operated for the US Department of Energy (DOE), filmed a series of crash and fire tests performed on three casks designed to transport irradiated nuclear fuel assemblies. While the tests were performed to assess the applicability of scale and computer modeling techniques to actual accidents, films of them were quickly pressed into service by the DOE and nuclear utilities as proof'' to the public of the safety of the casks. In the public debate over the safety of irradiated nuclear fuel transportation, the films have served asmore » the mainstay for the nuclear industry. Although the scripts of all the films were reviewed by USDOE officials before production, they contain numerous misleading concepts and images, and omit significant facts. The shorter versions eliminated qualifying statements contained in the longer version, and created false impressions. This paper discusses factors which cast doubt on the veracity of the films and the results of the tests.« less

Nuclear cask testing films misleading and misused

DOE Office of Scientific and Technical Information (OSTI.GOV)

Audin, L.

In 1977 and 1978, Sandia National Laboratories, located in Albuquerque, New Mexico, and operated for the US Department of Energy (DOE), filmed a series of crash and fire tests performed on three casks designed to transport irradiated nuclear fuel assemblies. While the tests were performed to assess the applicability of scale and computer modeling techniques to actual accidents, films of them were quickly pressed into service by the DOE and nuclear utilities as ``proof`` to the public of the safety of the casks. In the public debate over the safety of irradiated nuclear fuel transportation, the films have served asmore » the mainstay for the nuclear industry. Although the scripts of all the films were reviewed by USDOE officials before production, they contain numerous misleading concepts and images, and omit significant facts. The shorter versions eliminated qualifying statements contained in the longer version, and created false impressions. This paper discusses factors which cast doubt on the veracity of the films and the results of the tests.« less

Engineering bacteria to solve the Burnt Pancake Problem

PubMed Central

Haynes, Karmella A; Broderick, Marian L; Brown, Adam D; Butner, Trevor L; Dickson, James O; Harden, W Lance; Heard, Lane H; Jessen, Eric L; Malloy, Kelly J; Ogden, Brad J; Rosemond, Sabriya; Simpson, Samantha; Zwack, Erin; Campbell, A Malcolm; Eckdahl, Todd T; Heyer, Laurie J; Poet, Jeffrey L

2008-01-01

Background We investigated the possibility of executing DNA-based computation in living cells by engineering Escherichia coli to address a classic mathematical puzzle called the Burnt Pancake Problem (BPP). The BPP is solved by sorting a stack of distinct objects (pancakes) into proper order and orientation using the minimum number of manipulations. Each manipulation reverses the order and orientation of one or more adjacent objects in the stack. We have designed a system that uses site-specific DNA recombination to mediate inversions of genetic elements that represent pancakes within plasmid DNA. Results Inversions (or "flips") of the DNA fragment pancakes are driven by the Salmonella typhimurium Hin/hix DNA recombinase system that we reconstituted as a collection of modular genetic elements for use in E. coli. Our system sorts DNA segments by inversions to produce different permutations of a promoter and a tetracycline resistance coding region; E. coli cells become antibiotic resistant when the segments are properly sorted. Hin recombinase can mediate all possible inversion operations on adjacent flippable DNA fragments. Mathematical modeling predicts that the system reaches equilibrium after very few flips, where equal numbers of permutations are randomly sorted and unsorted. Semiquantitative PCR analysis of in vivo flipping suggests that inversion products accumulate on a time scale of hours or days rather than minutes. Conclusion The Hin/hix system is a proof-of-concept demonstration of in vivo computation with the potential to be scaled up to accommodate larger and more challenging problems. Hin/hix may provide a flexible new tool for manipulating transgenic DNA in vivo. PMID:18492232

Sorting nexin Snx41 is essential for conidiation and mediates glutathione-based antioxidant defense during invasive growth in Magnaporthe oryzae

PubMed Central

Deng, Yi Zhen; Qu, Ziwei; He, Yunlong; Naqvi, Naweed I.

2012-01-01

The sorting nexins Atg20/Snx42 and Snx41 regulate membrane traffic and endosomal protein sorting and are essential for Cvt and/or pexophagy in yeast. Previously, we showed that macroautophagy is necessary for conidiation in the rice-blast fungus Magnaporthe oryzae. Here, we analyzed the physiological function(s) of selective autophagy in Magnaporthe through targeted deletion of MGG_12832, an ortholog of yeast SNX41 and ATG20/SNX42. Loss of MGG_12832 (hereafter SNX41) abolished conidia formation and pathogenesis in M. oryzae. Snx41-GFP localized as dynamic puncta or short tubules that are partially associated with autophagosomes and/or autophagic vacuoles. PX domain, but not macroautophagy per se, was required for such localization of Snx41-GFP in Magnaporthe. Although not required for nonselective autophagy, Snx41 was essential for pexophagy in Magnaporthe. We identified Oxp1, an ATP-dependent oxoprolinase in the gamma-glutamyl cycle, as a binding partner and potential retrieval target of Snx41-dependent protein sorting. The substrate of Oxp1, 5-oxoproline, could partially restore conidiation in the snx41Δ. Exogenous glutathione, a product of the gamma-glutamyl cycle, significantly restored pathogenicity in the snx41Δ mutant, likely through counteracting the oxidative stress imposed by the host. We propose that the gamma-glutamyl cycle and glutathione biosynthesis are subject to regulation by Snx41-dependent vesicular trafficking, and mediate antioxidant defense crucial for in planta growth and pathogenic differentiation of Magnaporthe at the onset of blast disease in rice. PMID:22561104

Raft-mediated trafficking of apical resident proteins occurs in both direct and transcytotic pathways in polarized hepatic cells: role of distinct lipid microdomains.

PubMed

Slimane, Tounsia Aït; Trugnan, Germain; Van IJzendoorn, Sven C D; Hoekstra, Dick

2003-02-01

In polarized hepatic cells, pathways and molecular principles mediating the flow of resident apical bile canalicular proteins have not yet been resolved. Herein, we have investigated apical trafficking of a glycosylphosphatidylinositol-linked and two single transmembrane domain proteins on the one hand, and two polytopic proteins on the other in polarized HepG2 cells. We demonstrate that the former arrive at the bile canalicular membrane via the indirect transcytotic pathway, whereas the polytopic proteins reach the apical membrane directly, after Golgi exit. Most importantly, cholesterol-based lipid microdomains ("rafts") are operating in either pathway, and protein sorting into such domains occurs in the biosynthetic pathway, largely in the Golgi. Interestingly, rafts involved in the direct pathway are Lubrol WX insoluble but Triton X-100 soluble, whereas rafts in the indirect pathway are both Lubrol WX and Triton X-100 insoluble. Moreover, whereas cholesterol depletion alters raft-detergent insolubility in the indirect pathway without affecting apical sorting, protein missorting occurs in the direct pathway without affecting raft insolubility. The data implicate cholesterol as a traffic direction-determining parameter in the direct apical pathway. Furthermore, raft-cargo likely distinguishing single vs. multispanning membrane anchors, rather than rafts per se (co)determine the sorting pathway.

Raft-mediated Trafficking of Apical Resident Proteins Occurs in Both Direct and Transcytotic Pathways in Polarized Hepatic Cells: Role of Distinct Lipid Microdomains

PubMed Central

Slimane, Tounsia Aït; Trugnan, Germain; van IJzendoorn, Sven C.D.; Hoekstra, Dick

2003-01-01

In polarized hepatic cells, pathways and molecular principles mediating the flow of resident apical bile canalicular proteins have not yet been resolved. Herein, we have investigated apical trafficking of a glycosylphosphatidylinositol-linked and two single transmembrane domain proteins on the one hand, and two polytopic proteins on the other in polarized HepG2 cells. We demonstrate that the former arrive at the bile canalicular membrane via the indirect transcytotic pathway, whereas the polytopic proteins reach the apical membrane directly, after Golgi exit. Most importantly, cholesterol-based lipid microdomains (“rafts”) are operating in either pathway, and protein sorting into such domains occurs in the biosynthetic pathway, largely in the Golgi. Interestingly, rafts involved in the direct pathway are Lubrol WX insoluble but Triton X-100 soluble, whereas rafts in the indirect pathway are both Lubrol WX and Triton X-100 insoluble. Moreover, whereas cholesterol depletion alters raft-detergent insolubility in the indirect pathway without affecting apical sorting, protein missorting occurs in the direct pathway without affecting raft insolubility. The data implicate cholesterol as a traffic direction-determining parameter in the direct apical pathway. Furthermore, raft-cargo likely distinguishing single vs. multispanning membrane anchors, rather than rafts per se (co)determine the sorting pathway. PMID:12589058

Basolateral Sorting of Furin in MDCK Cells Requires a Phenylalanine-Isoleucine Motif Together with an Acidic Amino Acid Cluster

PubMed Central

Simmen, Thomas; Nobile, Massimo; Bonifacino, Juan S.; Hunziker, Walter

1999-01-01

Furin is a subtilisin-related endoprotease which processes a wide range of bioactive proteins. Furin is concentrated in the trans-Golgi network (TGN), where proteolytic activation of many precursor proteins takes place. A significant fraction of furin, however, cycles among the TGN, the plasma membrane, and endosomes, indicating that the accumulation in the TGN reflects a dynamic localization process. The cytosolic domain of furin is necessary and sufficient for TGN localization, and two signals are responsible for retrieval of furin to the TGN. A tyrosine-based (YKGL) motif mediates internalization of furin from the cell surface into endosomes. An acidic cluster that is part of two casein kinase II phosphorylation sites (SDSEEDE) is then responsible for retrieval of furin from endosomes to the TGN. In addition, the acidic EEDE sequence also mediates endocytic activity. Here, we analyzed the sorting of furin in polarized epithelial cells. We show that furin is delivered to the basolateral surface of MDCK cells, from where a significant fraction of the protein can return to the TGN. A phenylalanine-isoleucine motif together with the acidic EEDE cluster is required for basolateral sorting and constitutes a novel signal regulating intracellular traffic of furin. PMID:10082580

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rauch, Eric Benton

This report serves as a comprehensive overview of the Extended Storage of Used Nuclear Fuel work performed for the Material Protection, Accounting and Control Technologies campaign under the Department of Energy Office of Nuclear Energy. This paper describes a signature based on the source and fissile material distribution found within a population of used fuel assemblies combined with the neutron absorbers found within cask design that is unique to a specific cask with its specific arrangement of fuel. The paper describes all the steps used in producing and analyzing this signature from the beginning to the project end.

Proliferation resistance assessment of various methods of spent nuclear fuel storage and disposal

NASA Astrophysics Data System (ADS)

Kollar, Lenka

Many countries are planning to build or already are building new nuclear power plants to match their growing energy needs. Since all nuclear power plants handle nuclear materials that could potentially be converted and used for nuclear weapons, they each present a nuclear proliferation risk. Spent nuclear fuel presents the largest build-up of nuclear material at a power plant. This is a proliferation risk because spent fuel contains plutonium that can be chemically separated and used for a nuclear weapon. The International Atomic Energy Agency (IAEA) safeguards spent fuel in all non-nuclear weapons states that are party to the Non-Proliferation Treaty. Various safeguards methods are in use at nuclear power plants and research is underway to develop safeguards methods for spent fuel in centralized storage or underground storage and disposal. Each method of spent fuel storage presents different proliferation risks due to the nature of the storage method and the safeguards techniques that are utilized. Previous proliferation resistance and proliferation risk assessments have mainly compared nuclear material through the whole fuel cycle and not specifically focused on spent fuel storage. This project evaluates the proliferation resistance of the three main types of spent fuel storage: spent fuel pool, dry cask storage, and geological repository. The proliferation resistance assessment methodology that is used in this project is adopted from previous work and altered to be applicable to spent fuel storage. The assessment methodology utilizes various intrinsic and extrinsic proliferation-resistant attributes for each spent fuel storage type. These attributes are used to calculate a total proliferation resistant (PR) value. The maximum PR value is 1.00 and a greater number means that the facility is more proliferation resistant. Current data for spent fuel storage in the United States and around the world was collected. The PR values obtained from this data are 0.49 for the spent fuel pool, 0.42 for dry cask storage, 0.36 for the operating geological repository, and 0.28 for the closed geological repository. Therefore, the spent fuel pool is currently the most proliferation resistant method for storing spent fuel. The extrinsic attributes, mainly involving safeguards measures, affect the total PR value the most. As a result, several recommendations are made to improve the proliferation resistance of spent fuel. These recommendations include employing more advanced safeguards measures, such as verification techniques and remote monitoring, for dry cask storage and the geological repository. Dry cask storage facilities should also be located at the plant and in a secure building to minimize the proliferation risk. Finally, the cost-benefit analysis of increased safeguards needs to be considered. Taking these recommendations into account, the PR values of dry cask storage and the closed geological would be significantly increased, to 0.57 and 0.51, respectively. As a result, with increased safeguards to the safeguards level of the spent fuel pool, dry cask storage would be the most proliferation resistant method to store spent fuel. Therefore, the IAEA should continue to develop remote monitoring and cask storage verification techniques in order to improve the proliferation resistance of spent fuel.

Emerging roles of ARHGAP33 in intracellular trafficking of TrkB and pathophysiology of neuropsychiatric disorders

PubMed Central

Nakazawa, Takanobu; Hashimoto, Ryota; Sakoori, Kazuto; Sugaya, Yuki; Tanimura, Asami; Hashimotodani, Yuki; Ohi, Kazutaka; Yamamori, Hidenaga; Yasuda, Yuka; Umeda-Yano, Satomi; Kiyama, Yuji; Konno, Kohtarou; Inoue, Takeshi; Yokoyama, Kazumasa; Inoue, Takafumi; Numata, Shusuke; Ohnuma, Tohru; Iwata, Nakao; Ozaki, Norio; Hashimoto, Hitoshi; Watanabe, Masahiko; Manabe, Toshiya; Yamamoto, Tadashi; Takeda, Masatoshi; Kano, Masanobu

2016-01-01

Intracellular trafficking of receptor proteins is essential for neurons to detect various extracellular factors during the formation and refinement of neural circuits. However, the precise mechanisms underlying the trafficking of neurotrophin receptors to synapses remain elusive. Here, we demonstrate that a brain-enriched sorting nexin, ARHGAP33, is a new type of regulator for the intracellular trafficking of TrkB, a high-affinity receptor for brain-derived neurotrophic factor. ARHGAP33 knockout (KO) mice exhibit reduced expression of synaptic TrkB, impaired spine development and neuropsychiatric disorder-related behavioural abnormalities. These deficits are rescued by specific pharmacological enhancement of TrkB signalling in ARHGAP33 KO mice. Mechanistically, ARHGAP33 interacts with SORT1 to cooperatively regulate TrkB trafficking. Human ARHGAP33 is associated with brain phenotypes and reduced SORT1 expression is found in patients with schizophrenia. We propose that ARHGAP33/SORT1-mediated TrkB trafficking is essential for synapse development and that the dysfunction of this mechanism may be a new molecular pathology of neuropsychiatric disorders. PMID:26839058

ARF6 and GASP-1 are post-endocytic sorting proteins selectively involved in the intracellular trafficking of dopamine D2 receptors mediated by GRK and PKC in transfected cells

PubMed Central

Cho, DI; Zheng, M; Min, C; Kwon, KJ; Shin, CY; Choi, HK; Kim, KM

2013-01-01

Background and Purpose GPCRs undergo both homologous and heterologous regulatory processes in which receptor phosphorylation plays a critical role. The protein kinases responsible for each pathway are well established; however, other molecular details that characterize each pathway remain unclear. In this study, the molecular mechanisms that determine the differences in the functional roles and intracellular trafficking between homologous and PKC-mediated heterologous internalization pathways for the dopamine D2 receptor were investigated. Experimental Approach All of the S/T residues located within the intracellular loops of D2 receptor were mutated, and the residues responsible for GRK- and PKC-mediated internalization were determined in HEK-293 cells and SH-SY5Y cells. The functional role of receptor internalization and the cellular components that determine the post-endocytic fate of internalized D2 receptors were investigated in the transfected cells. Key Results T134, T225/S228/S229 and S325 were involved in PKC-mediated D2 receptor desensitization. S229 and adjacent S/T residues mediated the PKC-dependent internalization of D2 receptors, which induced down-regulation and desensitization. S/T residues within the second intracellular loop and T225 were the major residues involved in GRK-mediated internalization of D2 receptors, which induced receptor resensitization. ARF6 mediated the recycling of D2 receptors internalized in response to agonist stimulation. In contrast, GASP-1 mediated the down-regulation of D2 receptors internalized in a PKC-dependent manner. Conclusions and Implications GRK- and PKC-mediated internalizations of D2 receptors occur through different intracellular trafficking pathways and mediate distinct functional roles. Distinct S/T residues within D2 receptors and different sorting proteins are involved in the dissimilar regulation of D2 receptors by GRK2 and PKC. PMID:23082996

Mediation analysis allowing for exposure-mediator interactions and causal interpretation: theoretical assumptions and implementation with SAS and SPSS macros

PubMed Central

Valeri, Linda; VanderWeele, Tyler J.

2012-01-01

Mediation analysis is a useful and widely employed approach to studies in the field of psychology and in the social and biomedical sciences. The contributions of this paper are several-fold. First we seek to bring the developments in mediation analysis for non linear models within the counterfactual framework to the psychology audience in an accessible format and compare the sorts of inferences about mediation that are possible in the presence of exposure-mediator interaction when using a counterfactual versus the standard statistical approach. Second, the work by VanderWeele and Vansteelandt (2009, 2010) is extended here to allow for dichotomous mediators and count outcomes. Third, we provide SAS and SPSS macros to implement all of these mediation analysis techniques automatically and we compare the types of inferences about mediation that are allowed by a variety of software macros. PMID:23379553

Mediation analysis allowing for exposure-mediator interactions and causal interpretation: theoretical assumptions and implementation with SAS and SPSS macros.

PubMed

Valeri, Linda; Vanderweele, Tyler J

2013-06-01

Mediation analysis is a useful and widely employed approach to studies in the field of psychology and in the social and biomedical sciences. The contributions of this article are several-fold. First we seek to bring the developments in mediation analysis for nonlinear models within the counterfactual framework to the psychology audience in an accessible format and compare the sorts of inferences about mediation that are possible in the presence of exposure-mediator interaction when using a counterfactual versus the standard statistical approach. Second, the work by VanderWeele and Vansteelandt (2009, 2010) is extended here to allow for dichotomous mediators and count outcomes. Third, we provide SAS and SPSS macros to implement all of these mediation analysis techniques automatically, and we compare the types of inferences about mediation that are allowed by a variety of software macros. (PsycINFO Database Record (c) 2013 APA, all rights reserved).

The EARP Complex and Its Interactor EIPR-1 Are Required for Cargo Sorting to Dense-Core Vesicles

PubMed Central

Topalidou, Irini; Cattin-Ortolá, Jérôme; MacCoss, Michael J.

2016-01-01

The dense-core vesicle is a secretory organelle that mediates the regulated release of peptide hormones, growth factors, and biogenic amines. Dense-core vesicles originate from the trans-Golgi of neurons and neuroendocrine cells, but it is unclear how this specialized organelle is formed and acquires its specific cargos. To identify proteins that act in dense-core vesicle biogenesis, we performed a forward genetic screen in Caenorhabditis elegans for mutants defective in dense-core vesicle function. We previously reported the identification of two conserved proteins that interact with the small GTPase RAB-2 to control normal dense-core vesicle cargo-sorting. Here we identify several additional conserved factors important for dense-core vesicle cargo sorting: the WD40 domain protein EIPR-1 and the endosome-associated recycling protein (EARP) complex. By assaying behavior and the trafficking of dense-core vesicle cargos, we show that mutants that lack EIPR-1 or EARP have defects in dense-core vesicle cargo-sorting similar to those of mutants in the RAB-2 pathway. Genetic epistasis data indicate that RAB-2, EIPR-1 and EARP function in a common pathway. In addition, using a proteomic approach in rat insulinoma cells, we show that EIPR-1 physically interacts with the EARP complex. Our data suggest that EIPR-1 is a new interactor of the EARP complex and that dense-core vesicle cargo sorting depends on the EARP-dependent trafficking of cargo through an endosomal sorting compartment. PMID:27191843

FFTF disposable solid waste cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thomson, J. D.; Goetsch, S. D.

1983-01-01

Disposal of radioactive waste from the Fast Flux Test Facility (FFTF) will utilize a Disposable Solid Waste Cask (DSWC) for the transport and burial of irradiated stainless steel and inconel materials. Retrievability coupled with the desire for minimal facilities and labor costs at the disposal site identified the need for the DSWC. Design requirements for this system were patterned after Type B packages as outlined in 10 CFR 71 with a few exceptions based on site and payload requirements. A summary of the design basis, supporting analytical methods and fabrication practices developed to deploy the DSWC is provided in thismore » paper.« less

Array Detector Modules for Spent Fuel Verification

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bolotnikov, Aleksey

Brookhaven National Laboratory (BNL) proposes to evaluate the arrays of position-sensitive virtual Frisch-grid (VFG) detectors for passive gamma-ray emission tomography (ET) to verify the spent fuel in storage casks before storing them in geo-repositories. Our primary objective is to conduct a preliminary analysis of the arrays capabilities and to perform field measurements to validate the effectiveness of the proposed array modules. The outcome of this proposal will consist of baseline designs for the future ET system which can ultimately be used together with neutrons detectors. This will demonstrate the usage of this technology in spent fuel storage casks.

Neutron field characterization at the independent spent fuel storage installation of the Trillo nuclear power plant.

PubMed

Campo, Xandra; Méndez, Roberto; Embid, Miguel; Ortego, Alberto; Novo, Manuel; Sanz, Javier

2018-05-01

Neutron fields inside and outside the independent spent fuel storage installation of Trillo Nuclear Power Plant are characterized exhaustively in terms of neutron spectra and ambient dose equivalent, measured by Bonner sphere system and LB6411 monitor. Measurements are consistent with storage casks and building shield characteristics, and also with casks distribution inside the building. Outer values at least five times lower than dose limit for free access area are found. Measurements with LB6411 and spectrometer are consistent with each other. Copyright © 2018 Elsevier Ltd. All rights reserved.

Sorting nexin 9 differentiates ligand-activated Smad3 from Smad2 for nuclear import and transforming growth factor β signaling

PubMed Central

Wilkes, Mark C.; Repellin, Claire E.; Kang, Jeong-Han; Andrianifahanana, Mahefatiana; Yin, Xueqian; Leof, Edward B.

2015-01-01

Transforming growth factor β (TGFβ) is a pleiotropic protein secreted from essentially all cell types and primary tissues. While TGFβ’s actions reflect the activity of a number of signaling networks, the primary mediator of TGFβ responses are the Smad proteins. Following receptor activation, these cytoplasmic proteins form hetero-oligomeric complexes that translocate to the nucleus and affect gene transcription. Here, through biological, biochemical, and immunofluorescence approaches, sorting nexin 9 (SNX9) is identified as being required for Smad3-dependent responses. SNX9 interacts with phosphorylated (p) Smad3 independent of Smad2 or Smad4 and promotes more rapid nuclear delivery than that observed independent of ligand. Although SNX9 does not bind nucleoporins Nup153 or Nup214 or some β importins (Imp7 or Impβ), it mediates the association of pSmad3 with Imp8 and the nuclear membrane. This facilitates nuclear translocation of pSmad3 but not SNX9. PMID:26337383

Clathrin-independent carriers form a high capacity endocytic sorting system at the leading edge of migrating cells

PubMed Central

Howes, Mark T.; Kirkham, Matthew; Riches, James; Cortese, Katia; Walser, Piers J.; Simpson, Fiona; Hill, Michelle M.; Jones, Alun; Lundmark, Richard; Lindsay, Margaret R.; Hernandez-Deviez, Delia J.; Hadzic, Gordana; McCluskey, Adam; Bashir, Rumasia; Liu, Libin; Pilch, Paul; McMahon, Harvey; Robinson, Phillip J.; Hancock, John F.; Mayor, Satyajit

2010-01-01

Although the importance of clathrin- and caveolin-independent endocytic pathways has recently emerged, key aspects of these routes remain unknown. Using quantitative ultrastructural approaches, we show that clathrin-independent carriers (CLICs) account for approximately three times the volume internalized by the clathrin-mediated endocytic pathway, forming the major pathway involved in uptake of fluid and bulk membrane in fibroblasts. Electron tomographic analysis of the 3D morphology of the earliest carriers shows that they are multidomain organelles that form a complex sorting station as they mature. Proteomic analysis provides direct links between CLICs, cellular adhesion turnover, and migration. Consistent with this, CLIC-mediated endocytosis of key cargo proteins, CD44 and Thy-1, is polarized at the leading edge of migrating fibroblasts, while transient ablation of CLICs impairs their ability to migrate. These studies provide the first quantitative ultrastructural analysis and molecular characterization of the major endocytic pathway in fibroblasts, a pathway that provides rapid membrane turnover at the leading edge of migrating cells. PMID:20713605

The transcriptional control machinery as well as the cell wall integrity and its regulation are involved in the detoxification of the organic solvent dimethyl sulfoxide in Saccharomyces cerevisiae.

PubMed

Zhang, Lilin; Liu, Ningning; Ma, Xiao; Jiang, Linghuo

2013-03-01

In the present study, we have identified 339 dimethyl sulfoxide (DMSO)-sensitive and nine DMSO-tolerant gene mutations in Saccharomyces cerevisiae through a functional genomics approach. Twelve of these identified DMSO-sensitive mutations are of genes involved in the general control of gene expression mediated by the SWR1 complex and the RNA polymerase II mediator complex, whereas 71 of them are of genes involved in the protein trafficking and vacuolar sorting processes. In addition, twelve of these DMSO-sensitive mutations are of genes involved in the cell wall integrity (CWI) and its regulation. DMSO-tolerant mutations are of genes mainly involved in the metabolism and the gene expression control. Therefore, the transcriptional control machinery, the CWI and its regulation as well as the protein trafficking and sorting process play critical roles in the DMSO detoxification in yeast cells. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

mRNA localization: an orchestration of assembly, traffic and synthesis.

PubMed

Xing, Lei; Bassell, Gary J

2013-01-01

Asymmetrical mRNA localization and subsequent local translation provide efficient mechanisms for protein sorting in polarized cells. Defects in mRNA localization have been linked to developmental abnormalities and neurological diseases. Thus, it is critical to understand the machineries mediating and mechanisms underlying the asymmetrical distribution of mRNA and its regulation. The goal of this review is to summarize recent advances in the understanding of mRNA transport and localization, including the assembly and sorting of transport messenger ribonucleic protein (mRNP) granules, molecular mechanisms of active mRNP transport, cytoskeletal interactions and regulation of these events by extracellular signals. © 2012 John Wiley & Sons A/S.

Rab15 Effector Protein: A Novel Protein for Receptor Recycling from the Endocytic Recycling CompartmentD⃞

PubMed Central

Strick, David J.; Elferink, Lisa A.

2005-01-01

Sorting endosomes and the endocytic recycling compartment are critical intracellular stores for the rapid recycling of internalized membrane receptors to the cell surface in multiple cell types. However, the molecular mechanisms distinguishing fast receptor recycling from sorting endosomes and slow receptor recycling from the endocytic recycling compartment remain poorly understood. We previously reported that Rab15 differentially regulates transferrin receptor trafficking through sorting endosomes and the endocytic recycling compartment, suggesting a role for distinct Rab15-effector interactions at these endocytic compartments. In this study, we identified the novel protein Rab15 effector protein (REP15) as a binding partner for Rab15-GTP. REP15 is compartment specific, colocalizing with Rab15 and Rab11 on the endocytic recycling compartment but not with Rab15, Rab4, or early endosome antigen 1 on sorting endosomes. REP15 interacts directly with Rab15-GTP but not with Rab5 or Rab11. Consistent with its localization, REP15 overexpression and small interfering RNA-mediated depletion inhibited transferrin receptor recycling from the endocytic recycling compartment, without affecting receptor entry into or recycling from sorting endosomes. Our data identify REP15 as a compartment-specific protein for receptor recycling from the endocytic recycling compartment, highlighting that the rapid and slow modes of transferrin receptor recycling are mechanistically distinct pathways. PMID:16195351

Creation of a Geant4 Muon Tomography Package for Imaging of Nuclear Fuel in Dry Cask Storage

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tsoukalas, Lefteri H.

2016-03-01

This is the final report of the NEUP project “Creation of a Geant4 Muon Tomography Package for Imaging of Nuclear Fuel in Dry Cask Storage”, DE-NE0000695. The project started on December 1, 2013 and this report covers the period December 1, 2013 through November 30, 2015. The project was successfully completed and this report provides an overview of the main achievements, results and findings throughout the duration of the project. Additional details can be found in the main body of this report and on the individual Quarterly Reports and associated Deliverables of the project, uploaded in PICS-NE.

Present experience of NRI REZ with preparation of spent nuclear fuel shipment to Russian Federation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Svitak, F.; Broz, V.; Hrehor, M.

2008-07-15

The Nuclear Research Institute Rez plc (NRI) jointed the Russian Research Reactor Fuel Return (RRRFR) programme under the US-Russian Global Threat Reduction Initiative (GTRI) initiative and started the preparation of the spent nuclear fuel (SNF) shipment from the LVR-15 research reactor back to the Russian Federation (RF). The transport of 16 SKODA VPVR/M casks with EK-10, IRT-2M 80 %, and IRT-2M 36% fuel types is planned for the autumn of 2007. The paper describes the experience gained so far during the preparatory works for the SNF shipment (facility equipment modification, cask licenses) and the actual preparation of the SNF formore » transport, in particular its checking, repacking in a hot cell, loading into the VPVR/M casks, drying, manipulation, completion of the transport documentation, etc., including its transport to the SNF storage facility at the NRI before it is shipped to the RF. The paper also briefly describes a regulatory framework for these activities with a focus on legislative and methodological aspects of the return of vitrified waste back to the Czech Republic. (author)« less

Used fuel rail shock and vibration testing options analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ross, Steven B.; Best, Ralph E.; Klymyshyn, Nicholas A.

2014-09-25

The objective of the rail shock and vibration tests is to complete the framework needed to quantify loads of fuel assembly components that are necessary to guide materials research and establish a technical basis for review organizations such as the U.S. Nuclear Regulatory Commission (NRC). A significant body of experimental and numerical modeling data exists to quantify loads and failure limits applicable to normal conditions of transport (NCT) rail transport, but the data are based on assumptions that can only be verified through experimental testing. The test options presented in this report represent possible paths for acquiring the data thatmore » are needed to confirm the assumptions of previous work, validate modeling methods that will be needed for evaluating transported fuel on a case-by-case basis, and inform material test campaigns on the anticipated range of fuel loading. The ultimate goal of this testing is to close all of the existing knowledge gaps related to the loading of used fuel under NCT conditions and inform the experiments and analysis program on specific endpoints for their research. The options include tests that would use an actual railcar, surrogate assemblies, and real or simulated rail transportation casks. The railcar carrying the cradle, cask, and surrogate fuel assembly payload would be moved in a train operating over rail track modified or selected to impart shock and vibration forces that occur during normal rail transportation. Computer modeling would be used to help design surrogates that may be needed for a rail cask, a cask’s internal basket, and a transport cradle. The objective of the design of surrogate components would be to provide a test platform that effectively simulates responses to rail shock and vibration loads that would be exhibited by state-of-the-art rail cask, basket, and/or cradle structures. The computer models would also be used to help determine the placement of instrumentation (accelerometers and strain gauges) on the surrogate fuel assemblies, cask and cradle structures, and the railcar so that forces and deflections that would result in the greatest potential for damage to high burnup and long-cooled UNF can be determined. For purposes of this report we consider testing on controlled track when we have control of the track and speed to facilitate modeling.« less

Distinct Pathways Mediate the Sorting of Tail-anchored Mitochondrial Outer Membrane Proteins

USDA-ARS?s Scientific Manuscript database

Little is known about the biogenesis of tail-anchored (TA) proteins localized to the mitochondrial outer membrane in plant cells. To address this issue, we screened all of the (>500) known and predicted TA proteins in Arabidopsis for those annotated, based on Gene Ontology, to possess mitochondrial...

Drosophila Pkaap regulates Rab4/Rab11-dependent traffic and Rab11 exocytosis of innate immune cargo

PubMed Central

Sorvina, Alexandra; Shandala, Tetyana; Brooks, Douglas A.

2016-01-01

ABSTRACT The secretion of immune-mediators is a critical step in the host innate immune response to pathogen invasion, and Rab GTPases have an important role in the regulation of this process. Rab4/Rab11 recycling endosomes are involved in the sorting of immune-mediators into specialist Rab11 vesicles that can traffic this cargo to the plasma membrane; however, how this sequential delivery process is regulated has yet to be fully defined. Here, we report that Drosophila Pkaap, an orthologue of the human dual-specific A-kinase-anchoring protein 2 or D-AKAP2 (also called AKAP10), appeared to have a nucleotide-dependent localisation to Rab4 and Rab11 endosomes. RNAi silencing of pkaap altered Rab4/Rab11 recycling endosome morphology, suggesting that Pkaap functions in cargo sorting and delivery in the secretory pathway. The depletion of pkaap also had a direct effect on Rab11 vesicle exocytosis and the secretion of the antimicrobial peptide Drosomycin at the plasma membrane. We propose that Pkaap has a dual role in antimicrobial peptide traffic and exocytosis, making it an essential component for the secretion of inflammatory mediators and the defence of the host against pathogens. PMID:27190105

The vacuolar transport of aleurain-GFP and 2S albumin-GFP fusions is mediated by the same pre-vacuolar compartments in tobacco BY-2 and Arabidopsis suspension cultured cells.

PubMed

Miao, Yansong; Li, Kwun Yee; Li, Hong-Ye; Yao, Xiaoqiang; Jiang, Liwen

2008-12-01

Soluble proteins reach vacuoles because they contain vacuolar sorting determinants (VSDs) that are recognized by vacuolar sorting receptor (VSR) proteins. Pre-vacuolar compartments (PVCs), defined by VSRs and GFP-VSR reporters in tobacco BY-2 cells, are membrane-bound intermediate organelles that mediate protein traffic from the Golgi apparatus to the vacuole in plant cells. Multiple pathways have been demonstrated to be responsible for vacuolar transport of lytic enzymes and storage proteins to the lytic vacuole (LV) and the protein storage vacuole (PSV), respectively. However, the nature of PVCs for LV and PSV pathways remains unclear. Here, we used two fluorescent reporters, aleurain-GFP and 2S albumin-GFP, that represent traffic of lytic enzymes and storage proteins to LV and PSV, respectively, to study the PVC-mediated transport pathways via transient expression in suspension cultured cells. We demonstrated that the vacuolar transport of aleurain-GFP and 2S albumin-GFP was mediated by the same PVC populations in both tobacco BY-2 and Arabidopsis suspension cultured cells. These PVCs were defined by the seven GFP-AtVSR reporters. In wortmannin-treated cells, the vacuolated PVCs contained the mRFP-AtVSR reporter in their limiting membranes, whereas the soluble aleurain-GFP or 2S albumin-GFP remained in the lumen of the PVCs, indicating a possible in vivo relationship between receptor and cargo within PVCs.

Activity-dependent ubiquitination of GluA1 mediates a distinct AMPA receptor endocytosis and sorting pathway.

PubMed

Schwarz, Lindsay A; Hall, Benjamin J; Patrick, Gentry N

2010-12-08

The accurate trafficking of AMPA receptors (AMPARs) to and from the synapse is a critical component of learning and memory in the brain, whereas dysfunction of AMPAR trafficking is hypothesized to be an underlying mechanism of Alzheimer's disease. Previous work has shown that ubiquitination of integral membrane proteins is a common posttranslational modification used to mediate endocytosis and endocytic sorting of surface proteins in eukaryotic cells. Here we report that mammalian AMPARs become ubiquitinated in response to their activation. Using a mutant of GluA1 that is unable to be ubiquitinated at lysines on its C-terminus, we demonstrate that ubiquitination is required for internalization of surface AMPARs and their trafficking to the lysosome in response to the AMPAR agonist AMPA but not for internalization of AMPARs in response to the NMDA receptor agonist NMDA. Through overexpression or RNA interference-mediated knockdown, we identify that a specific E3 ligase, Nedd4-1 (neural-precursor cell-expressed developmentally downregulated gene 4-1), is necessary for this process. Finally, we show that ubiquitination of GluA1 by Nedd4-1 becomes more prevalent as neurons mature. Together, these data show that ubiquitination of GluA1-containing AMPARs by Nedd4-1 mediates their endocytosis and trafficking to the lysosome. Furthermore, these results provide insight into how hippocampal neurons regulate AMPAR trafficking and degradation with high specificity in response to differing neuronal signaling cues and suggest that changes to this pathway may occur as neurons mature.

Activity-Dependent Ubiquitination of GluA1 Mediates a Distinct AMPAR Endocytosis and Sorting Pathway

PubMed Central

Schwarz, Lindsay A.; Hall, Benjamin J.; Patrick, Gentry N.

2010-01-01

The accurate trafficking of AMPA receptors (AMPARs) to and from the synapse is a critical component of learning and memory in the brain, while dysfunction of AMPAR trafficking is hypothesized to be an underlying mechanism of Alzheimer’s disease. Previous work has shown that ubiquitination of integral membrane proteins is a common post-translational modification used to mediate endocytosis and endocytic sorting of surface proteins in eukaryotic cells. Here we report that mammalian AMPARs become ubiquitinated in response to their activation. Using a mutant of GluA1 that is unable to be ubiquitinated at lysines on its carboxy-terminus, we demonstrate that ubiquitination is required for internalization of surface AMPARs and their trafficking to the lysosome in response to the AMPAR agonist AMPA, but not for internalization of AMPARs in response to the NMDA receptor (NMDAR) agonist NMDA. Through over-expression or RNAi-mediated knockdown, we identify that a specific E3 ligase, Nedd4-1, is necessary for this process. Finally, we show that ubiquitination of GluA1 by Nedd4-1 becomes more prevalent as neurons mature. Together, these data show that ubiquitination of GluA1-containing AMPARs by Nedd4-1 mediates their endocytosis and trafficking to the lysosome. Furthermore, these results provide insight into how hippocampal neurons regulate AMPAR trafficking and degradation with high specificity in response to differing neuronal signaling cues, and suggest that changes to this pathway may occur as neurons mature. PMID:21148011

Cellular Auxin Homeostasis under High Temperature Is Regulated through a SORTING NEXIN1–Dependent Endosomal Trafficking Pathway[C][W

PubMed Central

Hanzawa, Taiki; Shibasaki, Kyohei; Numata, Takahiro; Kawamura, Yukio; Gaude, Thierry; Rahman, Abidur

2013-01-01

High-temperature-mediated adaptation in plant architecture is linked to the increased synthesis of the phytohormone auxin, which alters cellular auxin homeostasis. The auxin gradient, modulated by cellular auxin homeostasis, plays an important role in regulating the developmental fate of plant organs. Although the signaling mechanism that integrates auxin and high temperature is relatively well understood, the cellular auxin homeostasis mechanism under high temperature is largely unknown. Using the Arabidopsis thaliana root as a model, we demonstrate that under high temperature, roots counterbalance the elevated level of intracellular auxin by promoting shootward auxin efflux in a PIN-FORMED2 (PIN2)-dependent manner. Further analyses revealed that high temperature selectively promotes the retrieval of PIN2 from late endosomes and sorts them to the plasma membrane through an endosomal trafficking pathway dependent on SORTING NEXIN1. Our results demonstrate that recycling endosomal pathway plays an important role in facilitating plants adaptation to increased temperature. PMID:24003052

HID-1 controls formation of large dense core vesicles by influencing cargo sorting and trans-Golgi network acidification

PubMed Central

Hummer, Blake H.; de Leeuw, Noah F.; Burns, Christian; Chen, Lan; Joens, Matthew S.; Hosford, Bethany; Fitzpatrick, James A. J.; Asensio, Cedric S.

2017-01-01

Large dense core vesicles (LDCVs) mediate the regulated release of neuropeptides and peptide hormones. They form at the trans-Golgi network (TGN), where their soluble content aggregates to form a dense core, but the mechanisms controlling biogenesis are still not completely understood. Recent studies have implicated the peripheral membrane protein HID-1 in neuropeptide sorting and insulin secretion. Using CRISPR/Cas9, we generated HID-1 KO rat neuroendocrine cells, and we show that the absence of HID-1 results in specific defects in peptide hormone and monoamine storage and regulated secretion. Loss of HID-1 causes a reduction in the number of LDCVs and affects their morphology and biochemical properties, due to impaired cargo sorting and dense core formation. HID-1 KO cells also exhibit defects in TGN acidification together with mislocalization of the Golgi-enriched vacuolar H+-ATPase subunit isoform a2. We propose that HID-1 influences early steps in LDCV formation by controlling dense core formation at the TGN. PMID:29074564

Global Analysis of Yeast Endosomal Transport Identifies the Vps55/68 Sorting Complex

PubMed Central

Schluter, Cayetana; Lam, Karen K.Y.; Brumm, Jochen; Wu, Bella W.; Saunders, Matthew; Stevens, Tom H.

2008-01-01

Endosomal transport is critical for cellular processes ranging from receptor down-regulation and retroviral budding to the immune response. A full understanding of endosome sorting requires a comprehensive picture of the multiprotein complexes that orchestrate vesicle formation and fusion. Here, we use unsupervised, large-scale phenotypic analysis and a novel computational approach for the global identification of endosomal transport factors. This technique effectively identifies components of known and novel protein assemblies. We report the characterization of a previously undescribed endosome sorting complex that contains two well-conserved proteins with four predicted membrane-spanning domains. Vps55p and Vps68p form a complex that acts with or downstream of ESCRT function to regulate endosomal trafficking. Loss of Vps68p disrupts recycling to the TGN as well as onward trafficking to the vacuole without preventing the formation of lumenal vesicles within the MVB. Our results suggest the Vps55/68 complex mediates a novel, conserved step in the endosomal maturation process. PMID:18216282

Signal dependent transport of a membrane cargo from early endosomes to recycling endosomes.

PubMed

Mahmoud, Ismail S; Louber, Jade; Dower, Steve K; Verhagen, Anne M; Gleeson, Paul A

2017-08-01

Many membrane cargoes undergo endocytosis and intracellular recycling to the plasma membrane via the early endosomes and the recycling endosomes. However whether specific sorting signals are required for transport from early endosomes to recycling endosomes is not known and the current view is that transport to the recycling endosomes is by a passive default process. Here we show that the cytoplasmic tail of the neonatal Fc receptor (FcRn) contains discrete signals for endocytosis and for sorting to the recycling endosomes. The FcRn cytoplasmic tail has previously been shown to contain the unusual WISL motif for AP2/clathrin-mediated endocytosis. By analysing FcRn mutants and CD8/FcRn chimeric molecules, we have identified an extended WISL sequence (GLPAPWISL) which promotes sorting from the early endosomes to the recycling endosomes. The insertion of GLPAPWISL into the cytoplasmic tail of CD8 resulted in efficient endocytosis and trafficking to the recycling endosomes, with only low levels detected in the late endosomes. Replacement of the highly conserved GLAPAP sequence within the GLPAPWISL motif with alanine residues resulted in endocytosis of the CD8/FcRn chimera to the early endosomes which was then trafficked predominantly to the late endosomes rather than the recycling endosomes. These studies demonstrate that signals within the cytoplasmic domains of membrane cargo can mediate active transport from early to recycling endosomes. Copyright © 2017 Elsevier GmbH. All rights reserved.

Managing aging effects on dry cask storage systems for extended long-term storage and transportation of used fuel - rev. 0

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chopra, O.K.; Diercks, D.; Fabian, R.

The cancellation of the Yucca Mountain repository program in the United States raises the prospect of extended long-term storage (i.e., >120 years) and deferred transportation of used fuel at operating and decommissioned nuclear power plant sites. Under U.S. federal regulations contained in Title 10 of the Code of Federal Regulations (CFR) 72.42, the initial license term for an Independent Spent Fuel Storage Installation (ISFSI) must not exceed 40 years from the date of issuance. Licenses may be renewed by the U.S. Nuclear Regulatory Commission (NRC) at the expiration of the license term upon application by the licensee for a periodmore » not to exceed 40 years. Application for ISFSI license renewals must include the following: (1) Time-limited aging analyses (TLAAs) that demonstrate that structures, systems, and components (SSCs) important to safety will continue to perform their intended function for the requested period of extended operation; and (2) a description of the aging management program (AMP) for management of issues associated with aging that could adversely affect SSCs important to safety. In addition, the application must also include design bases information as documented in the most recent updated final safety analysis report as required by 10 CFR 72.70. Information contained in previous applications, statements, or reports filed with the Commission under the license may be incorporated by reference provided that those references are clear and specific. The NRC has recently issued the Standard Review Plan (SRP) for renewal of used-fuel dry cask storage system (DCSS) licenses and Certificates of Compliance (CoCs), NUREG-1927, under which NRC may renew a specific license or a CoC for a term not to exceed 40 years. Both the license and the CoC renewal applications must contain revised technical requirements and operating conditions (fuel storage, surveillance and maintenance, and other requirements) for the ISFSI and DCSS that address aging effects that could affect the safe storage of the used fuel. The information contained in the license and CoC renewal applications will require NRC review to verify that the aging effects on the SSCs in DCSSs/ ISFSIs are adequately managed for the period of extended operation. To date, all of the ISFSIs located across the United States with more than 1,500 dry casks loaded with used fuel have initial license terms of 20 years; three ISFSIs (Surry, H.B. Robinson and Oconee) have received their renewed licenses for 20 years, and two other ISFSIs (Calvert Cliffs and Prairie Island) have applied for license renewal for 40 years. This report examines issues related to managing aging effects on the SSCs in DCSSs/ISFSIs for extended long-term storage and transportation of used fuels, following an approach similar to that of the Generic Aging Lessons Learned (GALL) report, NUREG-1801, for the aging management and license renewal of nuclear power plants. The report contains five chapters and an appendix on quality assurance for aging management programs for used-fuel dry storage systems. Chapter I of the report provides an overview of the ISFSI license renewal process based on 10 CFR 72 and the guidance provided in NUREG-1927. Chapter II contains definitions and terms for structures and components in DCSSs, materials, environments, aging effects, and aging mechanisms. Chapter III and Chapter IV contain generic TLAAs and AMPs, respectively, that have been developed for managing aging effects on the SSCs important to safety in the dry cask storage system designs described in Chapter V. The summary descriptions and tabulations of evaluations of AMPs and TLAAs for the SSCs that are important to safety in Chapter V include DCSS designs (i.e., NUHOMS{reg_sign}, HI-STORM 100, Transnuclear (TN) metal cask, NAC International S/T storage cask, ventilated storage cask (VSC-24), and the Westinghouse MC-10 metal dry storage cask) that have been and continue to be used by utilities across the country for dry storage of used fuel to date. The goal of this report is to help establish the technical basis for extended long-term storage and transportation of used fuel.« less

Analysis for Eccentric Multi Canister Overpack (MCO) Drops at the Canister Storage Building (CSB) (CSB-S-0073)

DOE Office of Scientific and Technical Information (OSTI.GOV)

HOLLENBECK, R.G.

The Spent Nuclear Fuel (SNF) Canister Storage Building (CSB) is the interim storage facility for the K-Basin SNF at the US. Department of Energy (DOE) Hanford Site. The SNF is packaged in multi-canister overpacks (MCOs). The MCOs are placed inside transport casks, then delivered to the service station inside the CSB. At the service station, the MCO handling machine (MHM) moves the MCO from the cask to a storage tube or one of two sample/weld stations. There are 220 standard storage tubes and six overpack storage tubes in a below grade reinforced concrete vault. Each storage tube can hold twomore » MCOs.« less

Distinct Pathways Mediate the Sorting of Tail-anchored Mitochondrial Outer Membrane Proteins

USDA-ARS?s Scientific Manuscript database

Little is known about the biogenesis of tail-anchored (TA) proteins localized to the mitochondrial outer membrane in plant cells. To address this issue, we screened all of the (>600) known and predicted TA proteins in Arabidopsis thaliana for those annotated, based on Gene Ontology, to possess mitoc...

Active remodelling of the TIM23 complex during translocation of preproteins into mitochondria.

PubMed

Popov-Celeketić, Dusan; Mapa, Koyeli; Neupert, Walter; Mokranjac, Dejana

2008-05-21

The TIM23 (translocase of the mitochondrial inner membrane) complex mediates translocation of preproteins across and their insertion into the mitochondrial inner membrane. How the translocase mediates sorting of preproteins into the two different subcompartments is poorly understood. In particular, it is not clear whether association of two operationally defined parts of the translocase, the membrane-integrated part and the import motor, depends on the activity state of the translocase. We established conditions to in vivo trap the TIM23 complex in different translocation modes. Membrane-integrated part of the complex and import motor were always found in one complex irrespective of whether an arrested preprotein was present or not. Instead, we detected different conformations of the complex in response to the presence and, importantly, the type of preprotein being translocated. Two non-essential subunits of the complex, Tim21 and Pam17, modulate its activity in an antagonistic manner. Our data demonstrate that the TIM23 complex acts as a single structural and functional entity that is actively remodelled to sort preproteins into different mitochondrial subcompartments.

Active remodelling of the TIM23 complex during translocation of preproteins into mitochondria

PubMed Central

Popov-Čeleketić, Dus̆an; Mapa, Koyeli; Neupert, Walter; Mokranjac, Dejana

2008-01-01

The TIM23 (translocase of the mitochondrial inner membrane) complex mediates translocation of preproteins across and their insertion into the mitochondrial inner membrane. How the translocase mediates sorting of preproteins into the two different subcompartments is poorly understood. In particular, it is not clear whether association of two operationally defined parts of the translocase, the membrane-integrated part and the import motor, depends on the activity state of the translocase. We established conditions to in vivo trap the TIM23 complex in different translocation modes. Membrane-integrated part of the complex and import motor were always found in one complex irrespective of whether an arrested preprotein was present or not. Instead, we detected different conformations of the complex in response to the presence and, importantly, the type of preprotein being translocated. Two non-essential subunits of the complex, Tim21 and Pam17, modulate its activity in an antagonistic manner. Our data demonstrate that the TIM23 complex acts as a single structural and functional entity that is actively remodelled to sort preproteins into different mitochondrial subcompartments. PMID:18418384

Structural and functional insights into sorting nexin 5/6 interaction with bacterial effector IncE.

PubMed

Sun, Qingxiang; Yong, Xin; Sun, Xiaodong; Yang, Fan; Dai, Zhonghua; Gong, Yanqiu; Zhou, Liming; Zhang, Xia; Niu, Dawen; Dai, Lunzhi; Liu, Jia-Jia; Jia, Da

2017-01-01

The endosomal trafficking pathways are essential for many cellular activities. They are also important targets by many intracellular pathogens. Key regulators of the endosomal trafficking include the retromer complex and sorting nexins (SNXs). Chlamydia trachomatis effector protein IncE directly targets the retromer components SNX5 and SNX6 and suppresses retromer-mediated transport, but the exact mechanism has remained unclear. We present the crystal structure of the PX domain of SNX5 in complex with IncE, showing that IncE binds to a highly conserved hydrophobic groove of SNX5. The unique helical hairpin of SNX5/6 is essential for binding, explaining the specificity of SNX5/6 for IncE. The SNX5/6-IncE interaction is required for cellular localization of IncE and its inhibitory function. Mechanistically, IncE inhibits the association of CI-MPR cargo with retromer-containing endosomal subdomains. Our study provides new insights into the regulation of retromer-mediated transport and illustrates the intricate competition between host and pathogens in controlling cellular trafficking.

Cellular uptake and trafficking of polydiacetylene micelles

NASA Astrophysics Data System (ADS)

Gravel, Edmond; Thézé, Benoit; Jacques, Isabelle; Anilkumar, Parambath; Gombert, Karine; Ducongé, Frédéric; Doris, Eric

2013-02-01

Polydiacetylene (PDA) micelles coated with either carboxylate-, ammonium-, or methoxy-polyethyleneglycol (PEG) chains were assembled and loaded with a fluorescent dye (DiO). Their interaction with MCF-7 human breast tumor cells was investigated by epi-fluorescence microscopy and fluorescence-activated cell sorting (FACS) to determine their internalization pathway and intracellular fate. It was found that the ionic character of the micelles influenced their internalization kinetics through a caveolae-mediated pathway and that all micelle types behaved somewhat similarly inside cells.Polydiacetylene (PDA) micelles coated with either carboxylate-, ammonium-, or methoxy-polyethyleneglycol (PEG) chains were assembled and loaded with a fluorescent dye (DiO). Their interaction with MCF-7 human breast tumor cells was investigated by epi-fluorescence microscopy and fluorescence-activated cell sorting (FACS) to determine their internalization pathway and intracellular fate. It was found that the ionic character of the micelles influenced their internalization kinetics through a caveolae-mediated pathway and that all micelle types behaved somewhat similarly inside cells. Electronic supplementary information (ESI) available: Detailed synthetic procedures and supplementary figures. See DOI: 10.1039/c2nr34149b

Molecular mechanism to recruit galectin-3 into multivesicular bodies for polarized exosomal secretion.

PubMed

Bänfer, Sebastian; Schneider, Dominik; Dewes, Jenny; Strauss, Maximilian T; Freibert, Sven-A; Heimerl, Thomas; Maier, Uwe G; Elsässer, Hans-Peter; Jungmann, Ralf; Jacob, Ralf

2018-05-08

The beta-galactoside binding lectin galectin-3 (Gal3) is found intracellularly and in the extracellular space. Secretion of this lectin is mediated independently of the secretory pathway by a not yet defined nonclassical mechanism. Here, we found Gal3 in the lumen of exosomes. Superresolution and electron microscopy studies visualized Gal3 recruitment and sorting into intraluminal vesicles. Exosomal Gal3 release depends on the endosomal sorting complex required for transport I (ESCRT-I) component Tsg101 and functional Vps4a. Either Tsg101 knockdown or expression of dominant-negative Vps4a E228Q causes an intracellular Gal3 accumulation at multivesicular body formation sites. In addition, we identified a highly conserved tetrapeptide P(S/T)AP motif in the amino terminus of Gal3 that mediates a direct interaction with Tsg101. Mutation of the P(S/T)AP motif results in a loss of interaction and a dramatic decrease in exosomal Gal3 secretion. We conclude that Gal3 is a member of endogenous non-ESCRT proteins which are P(S/T)AP tagged for exosomal release.

Macrophage Sortilin Promotes LDL Uptake, Foam Cell Formation, and Atherosclerosis

PubMed Central

Patel, Kevin M.; Strong, Alanna; Tohyama, Junichiro; Jin, Xueting; Morales, Carlos R.; Billheimer, Jeffery; Millar, John; Kruth, Howard; Rader, Daniel J.

2015-01-01

Rationale Non-coding gene variants at the SORT1 locus are strongly associated with LDL-C levels as well as with coronary artery disease (CAD). SORT1 encodes a protein called sortilin, and hepatic sortilin modulates LDL metabolism by targeting apoB-containing lipoproteins to the lysosome. Sortilin is also expressed in macrophages, but its role in macrophage uptake of LDL and in atherosclerosis independent of plasma LDL-C levels is unknown. Objective To determine the effect of macrophage sortilin expression on LDL uptake, foam cell formation, and atherosclerosis. Methods and Results We crossed Sort1−/− mice onto a ‘humanized’ Apobec1−/−; hAPOB Tg background and determined that Sort1 deficiency on this background had no effect on plasma LDL-C levels but dramatically reduced atherosclerosis in the aorta and aortic root. In order to test whether this effect was a result of macrophage sortilin deficiency, we transplanted Sort1−/−;LDLR−/− or Sort1+/+;LDLR−/− bone marrow into Ldlr−/− mice and observed a similar reduction in atherosclerosis in mice lacking hematopoetic sortilin without an effect on plasma LDL-C levels. In an effort to determine the mechanism by which hematopoetic sortilin deficiency reduced atherosclerosis, we found no effect of sortilin deficiency on macrophage recruitment or LPS-induced cytokine release in vivo. In contrast, sortilin deficient macrophages had significantly reduced uptake of native LDL ex vivo and reduced foam cell formation in vivo, whereas sortilin overexpression in macrophages resulted in increased LDL uptake and foam cell formation. Conclusions Macrophage sortilin deficiency protects against atherosclerosis by reducing macrophage uptake of LDL. Sortilin-mediated uptake of native LDL into macrophages may be an important mechanism of foam cell formation and contributor to atherosclerosis development. PMID:25593281

Sorting of endocytosed transferrin and asialoglycoprotein occurs immediately after internalization in HepG2 cells

PubMed Central

1987-01-01

After receptor-mediated uptake, asialoglycoproteins are routed to lysosomes, while transferrin is returned to the medium as apotransferrin. This sorting process was analyzed using 3,3'- diaminobenzidine (DAB) cytochemistry, followed by Percoll density gradient cell fractionation. A conjugate of asialoorosomucoid (ASOR) and horseradish peroxidase (HRP) was used as a ligand for the asialoglycoprotein receptor. Cells were incubated at 0 degree C in the presence of both 131I-transferrin and 125I-ASOR/HRP. Endocytosis of prebound 125I-ASOR/HRP and 131I-transferrin was monitored by cell fractionation on Percoll density gradients. Incubation of the cell homogenate in the presence of DAB and H2O2 before cell fractionation gave rise to a density shift of 125I-ASOR/HRP-containing vesicles due to HRP-catalyzed DAB polymerization. An identical change in density for 125I-transferrin and 125I-ASOR/HRP, induced by DAB cytochemistry, is taken as evidence for the concomitant presence of both ligands in the same compartment. At 37 degrees C, sorting of the two ligands occurred with a half-time of approximately 2 min, and was nearly completed within 10 min. The 125I-ASOR/HRP-induced shift of 131I-transferrin was completely dependent on the receptor-mediated uptake of 125I-ASOR/HRP in the same compartment. In the presence of a weak base (0.3 mM primaquine), the recycling of transferrin receptors was blocked. The cell surface transferrin receptor population was decreased within 6 min to 15% of its original size. DAB cytochemistry showed that sorting between endocytosed 131I-transferrin and 125I-ASOR/HRP was also blocked in the presence of primaquine. These results indicate that transferrin and asialoglycoprotein are taken up via the same compartments and that segregation of the transferrin-receptor complex and asialoglycoprotein occurs very efficiently soon after uptake. PMID:3032986

Final Technical Report: Imaging a Dry Storage Cask with Cosmic Ray Muons

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yang, Haori; Hayward, Jason; Chichester, David

The goal of this project is to build a scaled prototype system for monitoring used nuclear fuel (UNF) dry storage casks (DSCs) through cosmic ray muon imaging. Such a system will have the capability of verifying the content inside a DSC without opening it. Because of the growth of the nuclear power industry in the U.S. and the policy decision to ban reprocessing of commercial UNF, the used fuel inventory at commercial reactor sites has been increasing. Currently, UNF needs to be moved to independent spent fuel storage installations (ISFSIs), as its inventory approaches the limit on capacity of on-sitemore » wet storage. Thereafter, the fuel will be placed in shipping containers to be transferred to a final disposal site. The ISFSIs were initially licensed as temporary facilities for ~20-yr periods. Given the cancellation of the Yucca mountain project and no clear path forward, extended dry-cask storage (~100 yr.) at ISFSIs is very likely. From the point of view of nuclear material protection, accountability and control technologies (MPACT) campaign, it is important to ensure that special nuclear material (SNM) in UNF is not stolen or diverted from civilian facilities for other use during the extended storage.« less

Development and Experimental Benchmark of Simulations to Predict Used Nuclear Fuel Cladding Temperatures during Drying and Transfer Operations

DOE Office of Scientific and Technical Information (OSTI.GOV)

Greiner, Miles

Radial hydride formation in high-burnup used fuel cladding has the potential to radically reduce its ductility and suitability for long-term storage and eventual transport. To avoid this formation, the maximum post-reactor temperature must remain sufficiently low to limit the cladding hoop stress, and so that hydrogen from the existing circumferential hydrides will not dissolve and become available to re-precipitate into radial hydrides under the slow cooling conditions during drying, transfer and early dry-cask storage. The objective of this research is to develop and experimentallybenchmark computational fluid dynamics simulations of heat transfer in post-pool-storage drying operations, when high-burnup fuel cladding ismore » likely to experience its highest temperature. These benchmarked tools can play a key role in evaluating dry cask storage systems for extended storage of high-burnup fuels and post-storage transportation, including fuel retrievability. The benchmarked tools will be used to aid the design of efficient drying processes, as well as estimate variations of surface temperatures as a means of inferring helium integrity inside the canister or cask. This work will be conducted effectively because the principal investigator has experience developing these types of simulations, and has constructed a test facility that can be used to benchmark them.« less

Phosphorylation on Ser-279 and Ser-282 of connexin43 regulates endocytosis and gap junction assembly in pancreatic cancer cells

PubMed Central

Johnson, Kristen E.; Mitra, Shalini; Katoch, Parul; Kelsey, Linda S.; Johnson, Keith R.; Mehta, Parmender P.

2013-01-01

The molecular mechanisms regulating the assembly of connexins (Cxs) into gap junctions are poorly understood. Using human pancreatic tumor cell lines BxPC3 and Capan-1, which express Cx26 and Cx43, we show that, upon arrival at the cell surface, the assembly of Cx43 is impaired. Connexin43 fails to assemble, because it is internalized by clathrin-mediated endocytosis. Assembly is restored upon expressing a sorting-motif mutant of Cx43, which does not interact with the AP2 complex, and by expressing mutants that cannot be phosphorylated on Ser-279 and Ser-282. The mutants restore assembly by preventing clathrin-mediated endocytosis of Cx43. Our results also document that the sorting-motif mutant is assembled into gap junctions in cells in which the expression of endogenous Cx43 has been knocked down. Remarkably, Cx43 mutants that cannot be phosphorylated on Ser-279 or Ser-282 are assembled into gap junctions only when connexons are composed of Cx43 forms that can be phosphorylated on these serines and forms in which phosphorylation on these serines is abolished. Based on the subcellular fate of Cx43 in single and contacting cells, our results document that the endocytic itinerary of Cx43 is altered upon cell–cell contact, which causes Cx43 to traffic by EEA1-negative endosomes en route to lysosomes. Our results further show that gap-junctional plaques formed of a sorting motif–deficient mutant of Cx43, which is unable to be internalized by the clathrin-mediated pathway, are predominantly endocytosed in the form of annular junctions. Thus the differential phosphorylation of Cx43 on Ser-279 and Ser-282 is fine-tuned to control Cx43’s endocytosis and assembly into gap junctions. PMID:23363606

Polarized release of T-cell-receptor-enriched microvesicles at the immunological synapse.

PubMed

Choudhuri, Kaushik; Llodrá, Jaime; Roth, Eric W; Tsai, Jones; Gordo, Susana; Wucherpfennig, Kai W; Kam, Lance C; Stokes, David L; Dustin, Michael L

2014-03-06

The recognition events that mediate adaptive cellular immunity and regulate antibody responses depend on intercellular contacts between T cells and antigen-presenting cells (APCs). T-cell signalling is initiated at these contacts when surface-expressed T-cell receptors (TCRs) recognize peptide fragments (antigens) of pathogens bound to major histocompatibility complex molecules (pMHC) on APCs. This, along with engagement of adhesion receptors, leads to the formation of a specialized junction between T cells and APCs, known as the immunological synapse, which mediates efficient delivery of effector molecules and intercellular signals across the synaptic cleft. T-cell recognition of pMHC and the adhesion ligand intercellular adhesion molecule-1 (ICAM-1) on supported planar bilayers recapitulates the domain organization of the immunological synapse, which is characterized by central accumulation of TCRs, adjacent to a secretory domain, both surrounded by an adhesive ring. Although accumulation of TCRs at the immunological synapse centre correlates with T-cell function, this domain is itself largely devoid of TCR signalling activity, and is characterized by an unexplained immobilization of TCR-pMHC complexes relative to the highly dynamic immunological synapse periphery. Here we show that centrally accumulated TCRs are located on the surface of extracellular microvesicles that bud at the immunological synapse centre. Tumour susceptibility gene 101 (TSG101) sorts TCRs for inclusion in microvesicles, whereas vacuolar protein sorting 4 (VPS4) mediates scission of microvesicles from the T-cell plasma membrane. The human immunodeficiency virus polyprotein Gag co-opts this process for budding of virus-like particles. B cells bearing cognate pMHC receive TCRs from T cells and initiate intracellular signals in response to isolated synaptic microvesicles. We conclude that the immunological synapse orchestrates TCR sorting and release in extracellular microvesicles. These microvesicles deliver transcellular signals across antigen-dependent synapses by engaging cognate pMHC on APCs.

Polarized release of T-cell-receptor-enriched microvesicles at the immunological synapse

NASA Astrophysics Data System (ADS)

Choudhuri, Kaushik; Llodrá, Jaime; Roth, Eric W.; Tsai, Jones; Gordo, Susana; Wucherpfennig, Kai W.; Kam, Lance C.; Stokes, David L.; Dustin, Michael L.

2014-03-01

The recognition events that mediate adaptive cellular immunity and regulate antibody responses depend on intercellular contacts between T cells and antigen-presenting cells (APCs). T-cell signalling is initiated at these contacts when surface-expressed T-cell receptors (TCRs) recognize peptide fragments (antigens) of pathogens bound to major histocompatibility complex molecules (pMHC) on APCs. This, along with engagement of adhesion receptors, leads to the formation of a specialized junction between T cells and APCs, known as the immunological synapse, which mediates efficient delivery of effector molecules and intercellular signals across the synaptic cleft. T-cell recognition of pMHC and the adhesion ligand intercellular adhesion molecule-1 (ICAM-1) on supported planar bilayers recapitulates the domain organization of the immunological synapse, which is characterized by central accumulation of TCRs, adjacent to a secretory domain, both surrounded by an adhesive ring. Although accumulation of TCRs at the immunological synapse centre correlates with T-cell function, this domain is itself largely devoid of TCR signalling activity, and is characterized by an unexplained immobilization of TCR-pMHC complexes relative to the highly dynamic immunological synapse periphery. Here we show that centrally accumulated TCRs are located on the surface of extracellular microvesicles that bud at the immunological synapse centre. Tumour susceptibility gene 101 (TSG101) sorts TCRs for inclusion in microvesicles, whereas vacuolar protein sorting 4 (VPS4) mediates scission of microvesicles from the T-cell plasma membrane. The human immunodeficiency virus polyprotein Gag co-opts this process for budding of virus-like particles. B cells bearing cognate pMHC receive TCRs from T cells and initiate intracellular signals in response to isolated synaptic microvesicles. We conclude that the immunological synapse orchestrates TCR sorting and release in extracellular microvesicles. These microvesicles deliver transcellular signals across antigen-dependent synapses by engaging cognate pMHC on APCs.

Rab1a regulates sorting of early endocytic vesicles

PubMed Central

Mukhopadhyay, Aparna; Quiroz, Jose A.

2014-01-01

We previously reported that Rab1a is associated with asialoorosomucoid (ASOR)-containing early endocytic vesicles, where it is required for their microtubule-based motility. In Rab1a knockdown (KD) cell lines, ASOR failed to segregate from its receptor and, consequently, did not reach lysosomes for degradation, indicating a defect in early endosome sorting. Although Rab1 is required for Golgi/endoplasmic reticulum trafficking, this process was unaffected, likely due to retained expression of Rab1b in these cells. The present study shows that Rab1a has a more general role in endocytic vesicle processing that extends to EGF and transferrin (Tfn) trafficking. Compared with results in control Huh7 cells, EGF accumulated in aggregates within Rab1a KD cells, failing to reach lysosomal compartments. Tfn, a prototypical example of recycling cargo, accumulated in a Rab11-mediated slow-recycling compartment in Rab1a KD cells, in contrast to control cells, which sort Tfn into a fast-recycling Rab4 compartment. These data indicate that Rab1a is an important regulator of early endosome sorting for multiple cargo species. The effectors and accessory proteins recruited by Rab1a to early endocytic vesicles include the minus-end-directed kinesin motor KifC1, while others remain to be discovered. PMID:24407591

Spatially restricted G protein-coupled receptor activity via divergent endocytic compartments.

PubMed

Jean-Alphonse, Frederic; Bowersox, Shanna; Chen, Stanford; Beard, Gemma; Puthenveedu, Manojkumar A; Hanyaloglu, Aylin C

2014-02-14

Postendocytic sorting of G protein-coupled receptors (GPCRs) is driven by their interactions between highly diverse receptor sequence motifs with their interacting proteins, such as postsynaptic density protein (PSD95), Drosophila disc large tumor suppressor (Dlg1), zonula occludens-1 protein (zo-1) (PDZ) domain proteins. However, whether these diverse interactions provide an underlying functional specificity, in addition to driving sorting, is unknown. Here we identify GPCRs that recycle via distinct PDZ ligand/PDZ protein pairs that exploit their recycling machinery primarily for targeted endosomal localization and signaling specificity. The luteinizing hormone receptor (LHR) and β2-adrenergic receptor (B2AR), two GPCRs sorted to the regulated recycling pathway, underwent divergent trafficking to distinct endosomal compartments. Unlike B2AR, which traffics to early endosomes (EE), LHR internalizes to distinct pre-early endosomes (pre-EEs) for its recycling. Pre-EE localization required interactions of the LHR C-terminal tail with the PDZ protein GAIP-interacting protein C terminus, inhibiting its traffic to EEs. Rerouting the LHR to EEs, or EE-localized GPCRs to pre-EEs, spatially reprograms MAPK signaling. Furthermore, LHR-mediated activation of MAPK signaling requires internalization and is maintained upon loss of the EE compartment. We propose that combinatorial specificity between GPCR sorting sequences and interacting proteins dictates an unprecedented spatiotemporal control in GPCR signal activity.

Spent fuel behavior under abnormal thermal transients during dry storage

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stahl, D.; Landow, M.P.; Burian, R.J.

1986-01-01

This study was performed to determine the effects of abnormally high temperatures on spent fuel behavior. Prior to testing, calculations using the CIRFI3 code were used to determine the steady-state fuel and cask component temperatures. The TRUMP code was used to determine transient heating rates under postulated abnormal events during which convection cooling of the cask surfaces was obstructed by a debris bed covering the cask. The peak rate of temperature rise during the first 6 h was calculated to be about 15/sup 0/C/h, followed by a rate of about 1/sup 0/C/h. A Turkey Point spent fuel rod segment wasmore » heated to approx. 800/sup 0/C. The segment deformed uniformly with an average strain of 17% at failure and a local strain of 60%. Pretest characterization of the spent fuel consisted of visual examination, profilometry, eddy-current examination, gamma scanning, fission gas collection, void volume measurement, fission gas analysis, hydrogen analysis of the cladding, burnup analysis, cladding metallography, and fuel ceramography. Post-test characterization showed that the failure was a pinhole cladding breach. The results of the tests showed that spent fuel temperatures in excess of 700/sup 0/C are required to produce a cladding breach in fuel rods pressurized to 500 psing (3.45 MPa) under postulated abnormal thermal transient cask conditions. The pinhole cladding breach that developed would be too small to compromise the confinement of spent fuel particles during an abnormal event or after normal cooling conditions are restored. This behavior is similar to that found in other slow ramp tests with irradiated and nonirradiated rod sections and nonirradiated whole rods under conditions that bracketed postulated abnormal heating rates. This similarity is attributed to annealing of the irradiation-strengthened Zircaloy cladding during heating. In both cases, the failure was a benign, ductile pinhole rupture.« less

Computational Fluid Dynamics Best Practice Guidelines in the Analysis of Storage Dry Cask

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zigh, A.; Solis, J.

2008-07-01

Computational fluid dynamics (CFD) methods are used to evaluate the thermal performance of a dry cask under long term storage conditions in accordance with NUREG-1536 [NUREG-1536, 1997]. A three-dimensional CFD model was developed and validated using data for a ventilated storage cask (VSC-17) collected by Idaho National Laboratory (INL). The developed Fluent CFD model was validated to minimize the modeling and application uncertainties. To address modeling uncertainties, the paper focused on turbulence modeling of buoyancy driven air flow. Similarly, in the application uncertainties, the pressure boundary conditions used to model the air inlet and outlet vents were investigated and validated.more » Different turbulence models were used to reduce the modeling uncertainty in the CFD simulation of the air flow through the annular gap between the overpack and the multi-assembly sealed basket (MSB). Among the chosen turbulence models, the validation showed that the low Reynolds k-{epsilon} and the transitional k-{omega} turbulence models predicted the measured temperatures closely. To assess the impact of pressure boundary conditions used at the air inlet and outlet channels on the application uncertainties, a sensitivity analysis of operating density was undertaken. For convergence purposes, all available commercial CFD codes include the operating density in the pressure gradient term of the momentum equation. The validation showed that the correct operating density corresponds to the density evaluated at the air inlet condition of pressure and temperature. Next, the validated CFD method was used to predict the thermal performance of an existing dry cask storage system. The evaluation uses two distinct models: a three-dimensional and an axisymmetrical representation of the cask. In the 3-D model, porous media was used to model only the volume occupied by the rodded region that is surrounded by the BWR channel box. In the axisymmetric model, porous media was used to model the entire region that encompasses the fuel assemblies as well as the gaps in between. Consequently, a larger volume is represented by porous media in the second model; hence, a higher frictional flow resistance is introduced in the momentum equations. The conservatism and the safety margins of these models were compared to assess the applicability and the realism of these two models. The three-dimensional model included fewer geometry simplifications and is recommended as it predicted less conservative fuel cladding temperature values, while still assuring the existence of adequate safety margins. (authors)« less

Discrete, continuous, and stochastic models of protein sorting in the Golgi apparatus

PubMed Central

Gong, Haijun; Guo, Yusong; Linstedt, Adam

2017-01-01

The Golgi apparatus plays a central role in processing and sorting proteins and lipids in eukaryotic cells. Golgi compartments constantly exchange material with each other and with other cellular components, allowing them to maintain and reform distinct identities despite dramatic changes in structure and size during cell division, development, and osmotic stress. We have developed three minimal models of membrane and protein exchange in the Golgi—a discrete, stochastic model, a continuous ordinary differential equation model, and a continuous stochastic differential equation model—each based on two fundamental mechanisms: vesicle-coat-mediated selective concentration of cargoes and soluble N-ethylmaleimide-sensitive factor attachment protein receptor SNARE proteins during vesicle formation and SNARE-mediated selective fusion of vesicles. By exploring where the models differ, we hope to discover whether the discrete, stochastic nature of vesicle-mediated transport is likely to have appreciable functional consequences for the Golgi. All three models show similar ability to restore and maintain distinct identities over broad parameter ranges. They diverge, however, in conditions corresponding to collapse and reassembly of the Golgi. The results suggest that a continuum model provides a good description of Golgi maintenance but that considering the discrete nature of vesicle-based traffic is important to understanding assembly and disassembly of the Golgi. Experimental analysis validates a prediction of the models that altering guanine nucleotide exchange factor expression levels will modulate Golgi size. PMID:20365406

Adaptor Protein Complex-2 (AP-2) and Epsin-1 Mediate Protease-activated Receptor-1 Internalization via Phosphorylation- and Ubiquitination-dependent Sorting Signals*

PubMed Central

Chen, Buxin; Dores, Michael R.; Grimsey, Neil; Canto, Isabel; Barker, Breann L.; Trejo, JoAnn

2011-01-01

Signaling by protease-activated receptor-1 (PAR1), a G protein-coupled receptor (GPCR) for thrombin, is regulated by desensitization and internalization. PAR1 desensitization is mediated by β-arrestins, like most classic GPCRs. In contrast, internalization of PAR1 occurs through a clathrin- and dynamin-dependent pathway independent of β-arrestins. PAR1 displays two modes of internalization. Constitutive internalization of unactivated PAR1 is mediated by the clathrin adaptor protein complex-2 (AP-2), where the μ2-adaptin subunit binds directly to a tyrosine-based motif localized within the receptor C-tail domain. However, AP-2 depletion only partially inhibits agonist-induced internalization of PAR1, suggesting a function for other clathrin adaptors in this process. Here, we now report that AP-2 and epsin-1 are both critical mediators of agonist-stimulated PAR1 internalization. We show that ubiquitination of PAR1 and the ubiquitin-interacting motifs of epsin-1 are required for epsin-1-dependent internalization of activated PAR1. In addition, activation of PAR1 promotes epsin-1 de-ubiquitination, which may increase its endocytic adaptor activity to facilitate receptor internalization. AP-2 also regulates activated PAR1 internalization via recognition of distal C-tail phosphorylation sites rather than the canonical tyrosine-based motif. Thus, AP-2 and epsin-1 are both required to promote efficient internalization of activated PAR1 and recognize discrete receptor sorting signals. This study defines a new pathway for internalization of mammalian GPCRs. PMID:21965661

Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation.

PubMed

Allen, Joselyn N; Dey, Adwitia; Nissly, Ruth; Fraser, James; Yu, Shan; Balandaram, Gayathri; Peters, Jeffrey M; Hankey-Giblin, Pamela A

2017-04-03

Obesity promotes a chronic inflammatory state that is largely mediated by tissue-resident macrophages as well as monocyte-derived macrophages. Diet-induced obesity (DIO) is a valuable model in studying the role of macrophage heterogeneity; however, adequate macrophage isolations are difficult to acquire from inflamed tissues. In this protocol, we outline the isolation steps and necessary troubleshooting guidelines derived from our studies for obtaining a suitable population of tissue-resident macrophages from mice following 18 weeks of high-fat (HFD) or high-fat/high-cholesterol (HFHCD) diet intervention. This protocol focuses on three hallmark tissues studied in obesity and atherosclerosis including the liver, white adipose tissues (WAT), and the aorta. We highlight how dualistic usage of flow cytometry can achieve a new dimension of isolation and characterization of tissue-resident macrophages. A fundamental section of this protocol addresses the intricacies underlying tissue-specific enzymatic digestions and macrophage isolation, and subsequent cell-surface antibody staining for flow cytometric analysis. This protocol addresses existing complexities underlying fluorescent-activated cell sorting (FACS) and presents clarifications to these complexities so as to obtain broad range characterization from adequately sorted cell populations. Alternate enrichment methods are included for sorting cells, such as the dense liver, allowing for flexibility and time management when working with FACS. In brief, this protocol aids the researcher to evaluate macrophage heterogeneity from a multitude of inflamed tissues in a given study and provides insightful troubleshooting tips that have been successful for favorable cellular isolation and characterization of immune cells in DIO-mediated inflammation.

VPS35 Mutations in Parkinson Disease

PubMed Central

Vilariño-Güell, Carles; Wider, Christian; Ross, Owen A.; Dachsel, Justus C.; Kachergus, Jennifer M.; Lincoln, Sarah J.; Soto-Ortolaza, Alexandra I.; Cobb, Stephanie A.; Wilhoite, Greggory J.; Bacon, Justin A.; Behrouz, Bahareh; Melrose, Heather L.; Hentati, Emna; Puschmann, Andreas; Evans, Daniel M.; Conibear, Elizabeth; Wasserman, Wyeth W.; Aasly, Jan O.; Burkhard, Pierre R.; Djaldetti, Ruth; Ghika, Joseph; Hentati, Faycal; Krygowska-Wajs, Anna; Lynch, Tim; Melamed, Eldad; Rajput, Alex; Rajput, Ali H.; Solida, Alessandra; Wu, Ruey-Meei; Uitti, Ryan J.; Wszolek, Zbigniew K.; Vingerhoets, François; Farrer, Matthew J.

2011-01-01

The identification of genetic causes for Mendelian disorders has been based on the collection of multi-incident families, linkage analysis, and sequencing of genes in candidate intervals. This study describes the application of next-generation sequencing technologies to a Swiss kindred presenting with autosomal-dominant, late-onset Parkinson disease (PD). The family has tremor-predominant dopa-responsive parkinsonism with a mean onset of 50.6 ± 7.3 years. Exome analysis suggests that an aspartic-acid-to-asparagine mutation within vacuolar protein sorting 35 (VPS35 c.1858G>A; p.Asp620Asn) is the genetic determinant of disease. VPS35 is a central component of the retromer cargo-recognition complex, is critical for endosome-trans-golgi trafficking and membrane-protein recycling, and is evolutionarily highly conserved. VPS35 c.1858G>A was found in all affected members of the Swiss kindred and in three more families and one patient with sporadic PD, but it was not observed in 3,309 controls. Further sequencing of familial affected probands revealed only one other missense variant, VPS35 c.946C>T; (p.Pro316Ser), in a pedigree with one unaffected and two affected carriers, and thus the pathogenicity of this mutation remains uncertain. Retromer-mediated sorting and transport is best characterized for acid hydrolase receptors. However, the complex has many types of cargo and is involved in a diverse array of biologic pathways from developmental Wnt signaling to lysosome biogenesis. Our study implicates disruption of VPS35 and retromer-mediated trans-membrane protein sorting, rescue, and recycling in the neurodegenerative process leading to PD. PMID:21763482

Used fuel storage monitoring using novel 4He scintillation fast neutron detectors and neutron energy discrimination analysis

NASA Astrophysics Data System (ADS)

Kelley, Ryan P.

With an increasing quantity of spent nuclear fuel being stored at power plants across the United States, the demand exists for a new method of cask monitoring. Certifying these casks for transportation and long-term storage is a unique dilemma: their sealed nature lends added security, but at the cost of requiring non-invasive measurement techniques to verify their contents. This research will design and develop a new method of passively scanning spent fuel casks using 4He scintillation detectors to make this process more accurate. 4He detectors are a relatively new technological development whose full capabilities have not yet been exploited. These detectors take advantage of the high 4He cross section for elastic scattering at fast neutron energies, particularly the resonance around 1 MeV. If one of these elastic scattering interactions occurs within the detector, the 4He nucleus takes energy from the incident neutron, then de-excites by scintillation. Photomultiplier Tubes (PMTs) at either end of the detector tube convert this emitted light into an electrical signal. The goal of this research is to use the neutron spectroscopy features of 4He scintillation detectors to maintain accountability of spent fuel in storage. This project will support spent fuel safeguards and the detection of fissile material, in order to minimize the risk of nuclear proliferation and terrorism.

Impact of modeling Choices on Inventory and In-Cask Criticality Calculations for Forsmark 3 BWR Spent Fuel

DOE Office of Scientific and Technical Information (OSTI.GOV)

Martinez-Gonzalez, Jesus S.; Ade, Brian J.; Bowman, Stephen M.

2015-01-01

Simulation of boiling water reactor (BWR) fuel depletion poses a challenge for nuclide inventory validation and nuclear criticality safety analyses. This challenge is due to the complex operating conditions and assembly design heterogeneities that characterize these nuclear systems. Fuel depletion simulations and in-cask criticality calculations are affected by (1) completeness of design information, (2) variability of operating conditions needed for modeling purposes, and (3) possible modeling choices. These effects must be identified, quantified, and ranked according to their significance. This paper presents an investigation of BWR fuel depletion using a complete set of actual design specifications and detailed operational datamore » available for five operating cycles of the Swedish BWR Forsmark 3 reactor. The data includes detailed axial profiles of power, burnup, and void fraction in a very fine temporal mesh for a GE14 (10×10) fuel assembly. The specifications of this case can be used to assess the impacts of different modeling choices on inventory prediction and in-cask criticality, specifically regarding the key parameters that drive inventory and reactivity throughout fuel burnup. This study focused on the effects of the fidelity with which power history and void fraction distributions are modeled. The corresponding sensitivity of the reactivity in storage configurations is assessed, and the impacts of modeling choices on decay heat and inventory are addressed.« less

Functional Disruption of the Netrin-1 Guidance Gue Leads to Disruption in Mammary Gland Development and Increased Tumor Incidence

DTIC Science & Technology

2005-07-01

interactions will be complicated by the observation that cues, such as Netrin ( Yebra et al., 2003) and a class 7, GPI-linked Semaphorin (Pasterkamp et al...In vivo cell sorting in complementary segmental domains mediated by Eph receptors and ephrins. Nature 399, 267-271. Yebra , M., Montgomery, A. M

Familiarity and Personal Experience as Mediators of Recall when Planning for Future Contingencies

ERIC Educational Resources Information Center

Klein, Stanley B.; Robertson, Theresa E.; Delton, Andrew W.; Lax, Moshe L.

2012-01-01

In this article, we demonstrate that planning tasks enhance recall when the context of planning (a) is self-referential and (b) draws on familiar scenarios represented in episodic memory. Specifically, we show that when planning tasks are sorted according to the degree to which they evoke memories of personally familiar scenarios (e.g., planning a…

PACS-1 Mediates Phosphorylation-Dependent Ciliary Trafficking of the CNG Channel in Olfactory Sensory Neurons

PubMed Central

Jenkins, Paul M.; Zhang, Lian; Thomas, Gary; Martens, Jeffrey R.

2009-01-01

Impaired ciliary protein transport in olfactory sensory neurons (OSNs) leads to anosmia, and is a newly recognized clinical manifestation of a class of human disorders called ciliopathies. Surprisingly little is known regarding the mechanisms controlling trafficking to this unique neuronal compartment. Here, we show a novel role for phosphofurin acidic cluster-sorting protein 1 (PACS-1) in the ciliary trafficking of the olfactory CNG channel. PACS-1 is an intracellular sorting protein that mediates its effects through the binding of acidic clusters on cargo protein. This interaction is dependent on CK2 phosphorylation of both PACS-1 and its cargo. We show that CNGB1b contains two putative PACS-1 binding sites, which are phosphorylated by the serine/threonine protein kinase, CK2. Additionally, we show that PACS-1 is expressed in OSNs and interacts in complex with the CNG channel. CK2 inhibition in native OSNs causes a loss of CNG channel from cilia and subsequent olfactory dysfunction, while adenoviral expression of mutant PACS-1 causes similar mislocalization. These results provide a mechanism for the subunit-dependent ciliary trafficking of the CNG channel and offer insight into the mechanisms of ciliary transport. PMID:19710307

PACS-1 mediates phosphorylation-dependent ciliary trafficking of the cyclic-nucleotide-gated channel in olfactory sensory neurons.

PubMed

Jenkins, Paul M; Zhang, Lian; Thomas, Gary; Martens, Jeffrey R

2009-08-26

Impaired ciliary protein transport in olfactory sensory neurons (OSNs) leads to anosmia, and is a newly recognized clinical manifestation of a class of human disorders called ciliopathies. Surprisingly little is known regarding the mechanisms controlling trafficking to this unique neuronal compartment. Here, we show a novel role for phosphofurin acidic cluster-sorting protein 1 (PACS-1) in the ciliary trafficking of the olfactory cyclic-nucleotide-gated (CNG) channel. PACS-1 is an intracellular sorting protein that mediates its effects through the binding of acidic clusters on cargo protein. This interaction is dependent on CK2 phosphorylation of both PACS-1 and its cargo. We show that CNGB1b contains two putative PACS-1 binding sites, which are phosphorylated by the serine/threonine protein kinase, CK2. Additionally, we show that PACS-1 is expressed in OSNs and interacts in complex with the CNG channel. CK2 inhibition in native OSNs causes a loss of CNG channel from cilia and subsequent olfactory dysfunction, while adenoviral expression of mutant PACS-1 causes similar mislocalization. These results provide a mechanism for the subunit-dependent ciliary trafficking of the CNG channel and offer insight into the mechanisms of ciliary transport.

SORLA regulates calpain-dependent degradation of synapsin.

PubMed

Hartl, Daniela; Nebrich, Grit; Klein, Oliver; Stephanowitz, Heike; Krause, Eberhard; Rohe, Michael

2016-09-01

Sorting-related receptor with A-type repeats (SORLA) is an intracellular sorting receptor in neurons and a major risk factor for Alzheimer disease. Here, we performed global proteome analyses in the brain of SORLA-deficient mice followed by biochemical and histopathologic studies to identify novel neuronal pathways affected by receptor dysfunction. We demonstrate that the lack of SORLA results in accumulation of phosphorylated synapsins in cortex and hippocampus. We propose an underlying molecular mechanism by demonstrating that SORLA interacts with phosphorylated synapsins through 14-3-3 adaptor proteins to deliver synapsins to calpain-mediated proteolytic degradation. Our results suggest a novel function for SORLA which is in control of synapsin degradation, potentially impacting on synaptic vesicle endocytosis and/or exocytosis. Copyright © 2016 The Alzheimer's Association. Published by Elsevier Inc. All rights reserved.

Effect of Control Blade History, and Axial Coolant Density and Burnup Profiles on BWR Burnup Credit

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ade, Brian J; Marshall, William BJ J; Martinez-Gonzalez, Jesus S

Oak Ridge National Laboratory (ORNL) and the US Nuclear Regulatory Commission (NRC) have initiated a multiyear project to investigate the application of burnup credit (BUC) for boiling water reactor (BWR) fuel in storage and transportation systems (often referred to as casks) and spent fuel pools (SFPs). This work is divided into two main phases. The first phase investigated the applicability of peak reactivity methods currently used in SFPs to transportation and storage casks and the validation of reactivity calculations and spent fuel compositions within these methods. The second phase focuses on extending BUC beyond peak reactivity. This paper documents themore » analysis of the effects of control blade insertion history, and moderator density and burnup axial profiles for extended BWR BUC.« less

EPRI/DOE High-Burnup Fuel Sister Rod Test Plan Simplification and Visualization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Saltzstein, Sylvia J.; Sorenson, Ken B.; Hanson, B. D.

The EPRI/DOE High-Burnup Confirmatory Data Project (herein called the “Demo”) is a multi-year, multi-entity test with the purpose of providing quantitative and qualitative data to show if high-burnup fuel mechanical properties change in dry storage over a ten-year period. The Demo involves obtaining 32 assemblies of high-burnup PWR fuel of common cladding alloys from the North Anna Nuclear Power Plant, loading them in an NRC-licensed TN-32B cask, drying them according to standard plant procedures, and then storing them on the North Anna dry storage pad for ten years. After the ten-year storage time, the cask will be opened and themore » mechanical properties of the rods will be tested and analyzed.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

This volume contains the interim change notice for the safety operation procedure for hot cell. It covers the master-slave manipulators, dry waste removal, cell transfers, hoists, cask handling, liquid waste system, and physical characterization of fluids.

Pleiotrophin suppression of receptor protein tyrosine phosphatase-β/ζ maintains the self-renewal competence of fetal human oligodendrocyte progenitor cells.

PubMed

McClain, Crystal R; Sim, Fraser J; Goldman, Steven A

2012-10-24

Oligodendrocyte progenitor cells (OPCs) persist in human white matter, yet the mechanisms by which they are maintained in an undifferentiated state are unknown. Human OPCs differentially express protein tyrosine phosphatase receptor β/ζ (PTPRZ1) and its inhibitory ligand, pleiotrophin, suggesting the maintenance of an autocrine loop by which PTPRZ1 activity is tonically suppressed. PTPRZ1 constitutively promotes the tyrosine dephosphorylation of β-catenin and, thus, β-catenin participation in T cell factor (TCF)-mediated transcription. Using CD140a/PDGFRα-based fluorescence-activated cell sorting to isolate fetal OPCs from the fetal brain at gestational ages 16-22 weeks, we asked whether pleiotrophin modulated the expansion of OPCs and, if so, whether this was effected through the serial engagement of PTPRZ1 and β-catenin-dependent signals, such as TCF-mediated transcription. Lentiviral shRNAi knockdown of PTPRZ1 induced TCF-mediated transcription and substantially augmented GSK3β inhibition-induced TCF-reporter luciferase expression, suggesting dual regulation of β-catenin and the importance of PTPRZ1 as a tonic brake upon TCF-dependent transcription. Pharmacological inhibition of GSK3β triggered substrate detachment and initiated sphere formation, yet had no effect on either proliferation or net cell number. In contrast, pleiotrophin strongly potentiated the proliferation of CD140a(+)-sorted OPCs, as did PTPRZ1 knockdown, which significantly increased the total number of population doublings exhibited by OPCs before mitotic senescence. These observations suggest that pleiotrophin inhibition of PTPRZ1 contributes to the homeostatic self-renewal of OPCs and that this process is mediated by the tonic activation of β-catenin/TCF-dependent transcription.

Pleiotrophin Suppression of Receptor Protein Tyrosine Phosphatase-β/ζ Maintains the Self-Renewal Competence of Fetal Human Oligodendrocyte Progenitor Cells

PubMed Central

McClain, Crystal R.; Sim, Fraser J.

2012-01-01

Oligodendrocyte progenitor cells (OPCs) persist in human white matter, yet the mechanisms by which they are maintained in an undifferentiated state are unknown. Human OPCs differentially express protein tyrosine phosphatase receptor β/ζ (PTPRZ1) and its inhibitory ligand, pleiotrophin, suggesting the maintenance of an autocrine loop by which PTPRZ1 activity is tonically suppressed. PTPRZ1 constitutively promotes the tyrosine dephosphorylation of β-catenin and, thus, β-catenin participation in T cell factor (TCF)-mediated transcription. Using CD140a/PDGFRα-based fluorescence-activated cell sorting to isolate fetal OPCs from the fetal brain at gestational ages 16–22 weeks, we asked whether pleiotrophin modulated the expansion of OPCs and, if so, whether this was effected through the serial engagement of PTPRZ1 and β-catenin-dependent signals, such as TCF-mediated transcription. Lentiviral shRNAi knockdown of PTPRZ1 induced TCF-mediated transcription and substantially augmented GSK3β inhibition-induced TCF-reporter luciferase expression, suggesting dual regulation of β-catenin and the importance of PTPRZ1 as a tonic brake upon TCF-dependent transcription. Pharmacological inhibition of GSK3β triggered substrate detachment and initiated sphere formation, yet had no effect on either proliferation or net cell number. In contrast, pleiotrophin strongly potentiated the proliferation of CD140a+-sorted OPCs, as did PTPRZ1 knockdown, which significantly increased the total number of population doublings exhibited by OPCs before mitotic senescence. These observations suggest that pleiotrophin inhibition of PTPRZ1 contributes to the homeostatic self-renewal of OPCs and that this process is mediated by the tonic activation of β-catenin/TCF-dependent transcription. PMID:23100427

27 CFR 26.206 - Marking packages and cases.

Code of Federal Regulations, 2010 CFR

2010-04-01

..., rectifier, or bottler shall serially number each case, barrel, cask, or similar container of distilled... distiller, rectifier, or bottler shall plainly print, stamp, or stencil with durable coloring material, in...

The Amphipathic Helix of Adenovirus Capsid Protein VI Contributes to Penton Release and Postentry Sorting

PubMed Central

Martinez, Ruben; Schellenberger, Pascale; Vasishtan, Daven; Aknin, Cindy; Austin, Sisley; Dacheux, Denis; Rayne, Fabienne; Siebert, Alistair; Ruzsics, Zsolt; Gruenewald, Kay

2014-01-01

ABSTRACT Nuclear delivery of the adenoviral genome requires that the capsid cross the limiting membrane of the endocytic compartment and traverse the cytosol to reach the nucleus. This endosomal escape is initiated upon internalization and involves a highly coordinated process of partial disassembly of the entering capsid to release the membrane lytic internal capsid protein VI. Using wild-type and protein VI-mutated human adenovirus serotype 5 (HAdV-C5), we show that capsid stability and membrane rupture are major determinants of entry-related sorting of incoming adenovirus virions. Furthermore, by using electron cryomicroscopy, as well as penton- and protein VI-specific antibodies, we show that the amphipathic helix of protein VI contributes to capsid stability by preventing premature disassembly and deployment of pentons and protein VI. Thus, the helix has a dual function in maintaining the metastable state of the capsid by preventing premature disassembly and mediating efficient membrane lysis to evade lysosomal targeting. Based on these findings and structural data from cryo-electron microscopy, we suggest a refined disassembly mechanism upon entry. IMPORTANCE In this study, we show the intricate connection of adenovirus particle stability and the entry-dependent release of the membrane-lytic capsid protein VI required for endosomal escape. We show that the amphipathic helix of the adenovirus internal protein VI is required to stabilize pentons in the particle while coinciding with penton release upon entry and that release of protein VI mediates membrane lysis, thereby preventing lysosomal sorting. We suggest that this dual functionality of protein VI ensures an optimal disassembly process by balancing the metastable state of the mature adenovirus particle. PMID:25473051

Assembly mechanism of FCT region type 1 pili in serotype M6 Streptococcus pyogenes.

PubMed

Nakata, Masanobu; Kimura, Keiji Richard; Sumitomo, Tomoko; Wada, Satoshi; Sugauchi, Akinari; Oiki, Eiji; Higashino, Miharu; Kreikemeyer, Bernd; Podbielski, Andreas; Okahashi, Nobuo; Hamada, Shigeyuki; Isoda, Ryutaro; Terao, Yutaka; Kawabata, Shigetada

2011-10-28

The human pathogen Streptococcus pyogenes produces diverse pili depending on the serotype. We investigated the assembly mechanism of FCT type 1 pili in a serotype M6 strain. The pili were found to be assembled from two precursor proteins, the backbone protein T6 and ancillary protein FctX, and anchored to the cell wall in a manner that requires both a housekeeping sortase enzyme (SrtA) and pilus-associated sortase enzyme (SrtB). SrtB is primarily required for efficient formation of the T6 and FctX complex and subsequent polymerization of T6, whereas proper anchoring of the pili to the cell wall is mainly mediated by SrtA. Because motifs essential for polymerization of pilus backbone proteins in other Gram-positive bacteria are not present in T6, we sought to identify the functional residues involved in this process. Our results showed that T6 encompasses the novel VAKS pilin motif conserved in streptococcal T6 homologues and that the lysine residue (Lys-175) within the motif and cell wall sorting signal of T6 are prerequisites for isopeptide linkage of T6 molecules. Because Lys-175 and the cell wall sorting signal of FctX are indispensable for substantial incorporation of FctX into the T6 pilus shaft, FctX is suggested to be located at the pilus tip, which was also implied by immunogold electron microscopy findings. Thus, the elaborate assembly of FCT type 1 pili is potentially organized by sortase-mediated cross-linking between sorting signals and the amino group of Lys-175 positioned in the VAKS motif of T6, thereby displaying T6 and FctX in a temporospatial manner.

Breaking bad habits by improving executive function in individuals with obesity.

PubMed

Allom, Vanessa; Mullan, Barbara; Smith, Evelyn; Hay, Phillipa; Raman, Jayanthi

2018-04-16

Two primary factors that contribute to obesity are unhealthy eating and sedentary behavior. These behaviors are particularly difficult to change in the long-term because they are often enacted habitually. Cognitive Remediation Therapy has been modified and applied to the treatment of obesity (CRT-O) with preliminary results of a randomized controlled trial demonstrating significant weight loss and improvements in executive function. The objective of this study was to conduct a secondary data analysis of the CRT-O trial to evaluate whether CRT-O reduces unhealthy habits that contribute to obesity via improvements in executive function. Eighty participants with obesity were randomized to CRT-O or control. Measures of executive function (Wisconsin Card Sort Task and Trail Making Task) and unhealthy eating and sedentary behavior habits were administered at baseline, post-intervention and at 3 month follow-up. Participants receiving CRT-O demonstrated improvements in both measures of executive function and reductions in both unhealthy habit outcomes compared to control. Mediation analyses revealed that change in one element of executive function performance (Wisconsin Card Sort Task perseverance errors) mediated the effect of CRT-O on changes in both habit outcomes. These results suggest that the effectiveness of CRT-O may result from the disruption of unhealthy habits made possible by improvements in executive function. In particular, it appears that cognitive flexibility, as measured by the Wisconsin Card Sort task, is a key mechanism in this process. Improving cognitive flexibility may enable individuals to capitalise on interruptions in unhealthy habits by adjusting their behavior in line with their weight loss goals rather than persisting with an unhealthy choice. The RCT was registered with the Australian New Zealand Registry of Clinical Trials (trial id: ACTRN12613000537752 ).

Role of adaptor proteins and clathrin in the trafficking of human kidney anion exchanger 1 (kAE1) to the cell surface.

PubMed

Junking, Mutita; Sawasdee, Nunghathai; Duangtum, Natapol; Cheunsuchon, Boonyarit; Limjindaporn, Thawornchai; Yenchitsomanus, Pa-thai

2014-07-01

Kidney anion exchanger 1 (kAE1) plays an important role in acid-base homeostasis by mediating chloride/bicarbornate (Cl-/HCO3-) exchange at the basolateral membrane of α-intercalated cells in the distal nephron. Impaired intracellular trafficking of kAE1 caused by mutations of SLC4A1 encoding kAE1 results in kidney disease - distal renal tubular acidosis (dRTA). However, it is not known how the intracellular sorting and trafficking of kAE1 from trans-Golgi network (TGN) to the basolateral membrane occurs. Here, we studied the role of basolateral-related sorting proteins, including the mu1 subunit of adaptor protein (AP) complexes, clathrin and protein kinase D, on kAE1 trafficking in polarized and non-polarized kidney cells. By using RNA interference, co-immunoprecipitation, yellow fluorescent protein-based protein fragment complementation assays and immunofluorescence staining, we demonstrated that AP-1 mu1A, AP-3 mu1, AP-4 mu1 and clathrin (but not AP-1 mu1B, PKD1 or PKD2) play crucial roles in intracellular sorting and trafficking of kAE1. We also demonstrated colocalization of kAE1 and basolateral-related sorting proteins in human kidney tissues by double immunofluorescence staining. These findings indicate that AP-1 mu1A, AP-3 mu1, AP-4 mu1 and clathrin are required for kAE1 sorting and trafficking from TGN to the basolateral membrane of acid-secreting α-intercalated cells. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Vascular effects of aldosterone: sorting out the receptors and the ligands.

PubMed

Feldman, Ross D; Gros, Robert

2013-12-01

Aldosterone has actions far beyond its role as a renal regulator of sodium reabsorption, and broader mechanisms of action than simply a transcriptional regulator. Aldosterone has a number of vascular effects, including regulation of vascular reactivity and vascular growth and/or development. Aldosterone-mediated effects on vascular reactivity reflect a balance between its endothelial-dependent vasodilator effects and its direct smooth muscle vasoconstrictor effects. The endothelial vasodilator effects of aldosterone are mediated by phosphatidylinositol 3-kinase-dependent activation of nitric oxide synthase. G-Protein oestrogen receptor (GPER) is a recently recognized G-protein coupled receptor (GPCR) that is activated by steroid hormones. It was first recognized as the GPCR mediating the rapid effects of oestrogens. Activation of GPER also mediates at least some of the vascular effects of aldosterone in smooth muscle and endothelial cells. In vascular endothelial cells, aldosterone activation of GPER mediates vasodilation. In contrast, activation of endothelial mineralocorticoid receptors has been linked to enhanced vasoconstrictor and/or impaired vasodilator responses. © 2013 Wiley Publishing Asia Pty Ltd.

Revisiting caveolin trafficking: the end of the caveosome

PubMed Central

Howes, Mark T.

2010-01-01

In this issue, a study by Hayer et al. (2010. J. Cell Biol. doi: 10.1083/jcb.201003086) provides insights into the trafficking of caveolins, the major membrane proteins of caveolae. As well as providing evidence for ubiquitin-mediated endosomal sorting and degradation of caveolin in multivesicular bodies (MVBs), the new findings question the existence of a unique organelle proposed nine years ago, the caveosome. PMID:21041440

InstrumentationPod (IPOD) User Guide

DOE Office of Scientific and Technical Information (OSTI.GOV)

Parker, Robert F.

2014-06-12

This document describes the Instrumentation Pod (IPOD) and its operation and use. The IPOD is a low-power detector system comprising a 3He tube with preamp for neutron detection, a microcontroller-based data acquisition system, a GPS receiver for locationdetermination and time-synchronization, and power filtering and protection. The IPOD is intended to be bolted to the top of Dual-Use Casks stored at Baikal-1 in Kazakhstan in order to maintain continuity of knowledge of the materials stored within the cask. The data acquisition system receives pulses from the neutron-detection preamp, combines this information with other sensor data, and stores the result on twomore » SD cards that are part of the data acquisition system. Firmware in the data acquisition system controls collection and storing of the data and enables configuration of the acquisition parameters.« less

Development of Friction Stir Processing for Repair of Nuclear Dry Cask Storage System Canisters

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ross, Kenneth A.; Sutton, Ben; Grant, Glenn J.

The Nuclear Regulatory Commission has identified chloride-induced stress corrosion cracking (CISCC) of austenitic stainless steel dry cask storage systems (DCSS) as an area of great concern. Friction Stir Processing (FSP) was used to repair laboratory-generated stress corrosion cracking (SCC) in representative stainless steel 304 coupons. Results of this study show FSP is a viable method for repair and mitigation CISCC. This paper highlights lessons learned and developed techniques relative to FSP development for crack repair in sensitized thick section stainless steel 304. These include: development of process parameters, welding at low spindle speed, use of weld power and temperature controlmore » and optimization of these controls. NDE and destructive analysis are also presented to demonstrate effectiveness of the developed methods for SCC crack repair.« less

Horizontal modular dry irradiated fuel storage system

DOEpatents

Fischer, Larry E.; McInnes, Ian D.; Massey, John V.

1988-01-01

A horizontal, modular, dry, irradiated fuel storage system (10) includes a thin-walled canister (12) for containing irradiated fuel assemblies (20), which canister (12) can be positioned in a transfer cask (14) and transported in a horizontal manner from a fuel storage pool (18), to an intermediate-term storage facility. The storage system (10) includes a plurality of dry storage modules (26) which accept the canister (12) from the transfer cask (14) and provide for appropriate shielding about the canister (12). Each module (26) also provides for air cooling of the canister (12) to remove the decay heat of the irradiated fuel assemblies (20). The modules (26) can be interlocked so that each module (26) gains additional shielding from the next adjacent module (26). Hydraulic rams (30) are provided for inserting and removing the canisters (12) from the modules (26).

The used nuclear fuel problem - can reprocessing and consolidated storage be complementary?

DOE Office of Scientific and Technical Information (OSTI.GOV)

Phillips, C.; Thomas, I.

2013-07-01

This paper describes our CISF (Consolidated Interim Storage Facilities) and Reprocessing Facility concepts and show how they can be combined with a geologic repository to provide a comprehensive system for dealing with spent fuels in the USA. The performance of the CISF was logistically analyzed under six operational scenarios. A 3-stage plan has been developed to establish the CISF. Stage 1: the construction at the CISF site of only a rail receipt interface and storage pad large enough for the number of casks that will be received. The construction of the CISF Canister Handling Facility, the Storage Cask Fabrication Facility,more » the Cask Maintenance Facility and supporting infrastructure are performed during stage 2. The construction and placement into operation of a water-filled pool repackaging facility is completed for Stage 3. By using this staged approach, the capital cost of the CISF is spread over a number of years. It also allows more time for a final decision on the geologic repository to be made. A recycling facility will be built, this facility will used the NUEX recycling process that is based on the aqueous-based PUREX solvent extraction process, using a solvent of tri-N-butyl phosphate in a kerosene diluent. It is capable of processing spent fuels at a rate of 5 MT per day, at burn-ups up to 50 GWD per ton of spent fuels and a minimum of 5 years out-of-reactor cooling.« less

Adsorbed radioactivity and radiographic imaging of surfaces of stainless steel and titanium

NASA Astrophysics Data System (ADS)

Jung, Haijo

1997-11-01

Type 304 stainless steel used for typical surface materials of spent fuel shipping casks and titanium were exposed in the spent fuel storage pool of a typical PWR power plant. Adsorption characteristics, effectiveness of decontamination by water cleaning and by electrocleaning, and swipe effectiveness on the metal surfaces were studied. A variety of environmental conditions had been manipulated to stimulate the potential 'weeping' phenomenon that often occurs with spent fuel shipping casks during transit. In a previous study, few heterogeneous effects of adsorbed contamination onto metal surfaces were observed. Radiographic images of cask surfaces were made in this study and showed clearly heterogeneous activity distributions. Acquired radiographic images were digitized and further analyzed with an image analysis computer package and compared to calibrated images by using standard sources. The measurements of activity distribution by using the radiographic image method were consistent with that using a HPGe detector. This radiographic image method was used to study the effects of electrocleaning for total and specified areas. The Modulation Transfer Function (MTF) of a film-screen system in contact with a radioactive metal surface was studied with neutron activated gold foils and showed more broad resolution properties than general diagnostic x-ray film-screen systems. Microstructure between normal areas and hot spots showed significant differences, and one hot spot appearing as a dot on the film image consisted of several small hot spots (about 10 μm in diameter). These hot spots were observed as structural defects of the metal surfaces.

The C-Terminal Sequence of RhoB Directs Protein Degradation through an Endo-Lysosomal Pathway

PubMed Central

Ramos, Irene; Herrera, Mónica; Stamatakis, Konstantinos

2009-01-01

Background Protein degradation is essential for cell homeostasis. Targeting of proteins for degradation is often achieved by specific protein sequences or posttranslational modifications such as ubiquitination. Methodology/Principal Findings By using biochemical and genetic tools we have monitored the localization and degradation of endogenous and chimeric proteins in live primary cells by confocal microscopy and ultra-structural analysis. Here we identify an eight amino acid sequence from the C-terminus of the short-lived GTPase RhoB that directs the rapid degradation of both RhoB and chimeric proteins bearing this sequence through a lysosomal pathway. Elucidation of the RhoB degradation pathway unveils a mechanism dependent on protein isoprenylation and palmitoylation that involves sorting of the protein into multivesicular bodies, mediated by the ESCRT machinery. Moreover, RhoB sorting is regulated by late endosome specific lipid dynamics and is altered in human genetic lipid traffic disease. Conclusions/Significance Our findings characterize a short-lived cytosolic protein that is degraded through a lysosomal pathway. In addition, we define a novel motif for protein sorting and rapid degradation, which allows controlling protein levels by means of clinically used drugs. PMID:19956591

75 FR 25120 - List of Approved Spent Fuel Storage Casks: NUHOMS® HD System Revision 1

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-07

...-235, clarify the requirements of reconstituted fuel assemblies, add requirements to qualify metal matrix composite neutron absorbers with integral aluminum cladding, delete use of nitrogen for draining...

10 CFR 72.236 - Specific requirements for spent fuel storage cask approval and fabrication.

Code of Federal Regulations, 2013 CFR

2013-01-01

... maintained in a subcritical condition under credible conditions. (d) Radiation shielding and confinement... confinement of radioactive material under normal, off-normal, and credible accident conditions. (m) To the...

10 CFR 72.236 - Specific requirements for spent fuel storage cask approval and fabrication.

Code of Federal Regulations, 2012 CFR

2012-01-01

... maintained in a subcritical condition under credible conditions. (d) Radiation shielding and confinement... confinement of radioactive material under normal, off-normal, and credible accident conditions. (m) To the...

10 CFR 72.236 - Specific requirements for spent fuel storage cask approval and fabrication.

Code of Federal Regulations, 2010 CFR

2010-01-01

... maintained in a subcritical condition under credible conditions. (d) Radiation shielding and confinement... of radioactive material under normal, off-normal, and credible accident conditions. (m) To the extent...

10 CFR 72.236 - Specific requirements for spent fuel storage cask approval and fabrication.

Code of Federal Regulations, 2011 CFR

2011-01-01

... maintained in a subcritical condition under credible conditions. (d) Radiation shielding and confinement... of radioactive material under normal, off-normal, and credible accident conditions. (m) To the extent...

10 CFR 72.236 - Specific requirements for spent fuel storage cask approval and fabrication.

Code of Federal Regulations, 2014 CFR

2014-01-01

... maintained in a subcritical condition under credible conditions. (d) Radiation shielding and confinement... confinement of radioactive material under normal, off-normal, and credible accident conditions. (m) To the...

Sorting of Clathrin-Independent Cargo Proteins Depends on Rab35 Delivered by Clathrin-Mediated Endocytosis.

PubMed

Dutta, Dipannita; Donaldson, Julie G

2015-09-01

Clathrin-mediated endocytosis (CME) and clathrin-independent endocytosis (CIE) co-exist in most cells but little is known about their communication and coordination. Here we show that when CME was inhibited, endocytosis by CIE continued but endosomal trafficking of CIE cargo proteins was altered. CIE cargo proteins that normally traffic directly into Arf6-associated tubules after internalization and avoid degradation (CD44, CD98 and CD147) now trafficked to lysosomes and were degraded. The endosomal tubules were also absent and Arf6-GTP levels were elevated. The altered trafficking, loss of the tubular endosomal network and elevated Arf6-GTP levels caused by inhibition of CME were rescued by expression of Rab35, a Rab associated with clathrin-coated vesicles, or its effector ACAPs, Arf6 GTPase activating proteins (GAP) that inactivate Arf6. Furthermore, siRNA knockdown of Rab35 recreated the phenotype of CME ablation on CIE cargo trafficking without altering endocytosis of transferrin. These observations suggest that Rab35 serves as a CME detector and that loss of CME, or Rab35 input, leads to elevated Arf6-GTP and shifts the sorting of CIE cargo proteins to lysosomes and degradation. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.

Knowns and unknowns of plasma membrane protein degradation in plants.

PubMed

Liu, Chuanliang; Shen, Wenjin; Yang, Chao; Zeng, Lizhang; Gao, Caiji

2018-07-01

Plasma membrane (PM) not only creates a physical barrier to enclose the intracellular compartments but also mediates the direct communication between plants and the ever-changing environment. A tight control of PM protein homeostasis by selective degradation is thus crucial for proper plant development and plant-environment interactions. Accumulated evidences have shown that a number of plant PM proteins undergo clathrin-dependent or membrane microdomain-associated endocytic routes to vacuole for degradation in a cargo-ubiquitination dependent or independent manner. Besides, several trans-acting determinants involved in the regulation of endocytosis, recycling and multivesicular body-mediated vacuolar sorting have been identified in plants. More interestingly, recent findings have uncovered the participation of selective autophagy in PM protein turnover in plants. Although great progresses have been made to identify the PM proteins that undergo dynamic changes in subcellular localizations and to explore the factors that control the membrane protein trafficking, several questions remain to be answered regarding the molecular mechanisms of PM protein degradation in plants. In this short review article, we briefly summarize recent progress in our understanding of the internalization, sorting and degradation of plant PM proteins. More specifically, we focus on discussing the elusive aspects underlying the pathways of PM protein degradation in plants. Copyright © 2018 Elsevier B.V. All rights reserved.

Management of spent nuclear fuel on the Oak Ridge Reservation, Oak Ridge, Tennessee: Environmental assessment

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1996-02-01

On June 1, 1995, DOE issued a Record of Decision [60 Federal Register 28680] for the Department-wide management of spent nuclear fuel (SNF); regionalized storage of SNF by fuel type was selected as the preferred alternative. The proposed action evaluated in this environmental assessment is the management of SNF on the Oak Ridge Reservation (ORR) to implement this preferred alternative of regional storage. SNF would be retrieved from storage, transferred to a hot cell if segregation by fuel type and/or repackaging is required, loaded into casks, and shipped to off-site storage. The proposed action would also include construction and operationmore » of a dry cask SNF storage facility on ORR, in case of inadequate SNF storage. Action is needed to enable DOE to continue operation of the High Flux Isotope Reactor, which generates SNF. This report addresses environmental impacts.« less

A network of autism linked genes stabilizes two pools of synaptic GABAA receptors

PubMed Central

Tong, Xia-Jing; Hu, Zhitao; Liu, Yu; Anderson, Dorian; Kaplan, Joshua M

2015-01-01

Changing receptor abundance at synapses is an important mechanism for regulating synaptic strength. Synapses contain two pools of receptors, immobilized and diffusing receptors, both of which are confined to post-synaptic elements. Here we show that immobile and diffusing GABAA receptors are stabilized by distinct synaptic scaffolds at C. elegans neuromuscular junctions. Immobilized GABAA receptors are stabilized by binding to FRM-3/EPB4.1 and LIN-2A/CASK. Diffusing GABAA receptors are stabilized by the synaptic adhesion molecules Neurexin and Neuroligin. Inhibitory post-synaptic currents are eliminated in double mutants lacking both scaffolds. Neurexin, Neuroligin, and CASK mutations are all linked to Autism Spectrum Disorders (ASD). Our results suggest that these mutations may directly alter inhibitory transmission, which could contribute to the developmental and cognitive deficits observed in ASD. DOI: http://dx.doi.org/10.7554/eLife.09648.001 PMID:26575289

Bioremediation of Mercury by Vibrio fluvialis Screened from Industrial Effluents

PubMed Central

Saranya, Kailasam; Shekhar, Sudhanshu; Swaminathan, Sankaran; Balasubramanian, Thangavel

2017-01-01

Thirty-one mercury-resistant bacterial strains were isolated from the effluent discharge sites of the SIPCOT industrial area. Among them, only one strain (CASKS5) was selected for further investigation due to its high minimum inhibitory concentration of mercury and low antibiotic susceptibility. In accordance with 16S ribosomal RNA gene sequences, the strain CASKS5 was identified as Vibrio fluvialis. The mercury-removal capacity of V. fluvialis was analyzed at four different concentrations (100, 150, 200, and 250 μg/ml). Efficient bioremediation was observed at a level of 250 μg/ml with the removal of 60% of mercury ions. The interesting outcome of this study was that the strain V. fluvialis had a high bioremediation efficiency but had a low antibiotic resistance. Hence, V. fluvialis could be successfully used as a strain for the ecofriendly removal of mercury. PMID:28626761

Bioremediation of Mercury by Vibrio fluvialis Screened from Industrial Effluents.

PubMed

Saranya, Kailasam; Sundaramanickam, Arumugam; Shekhar, Sudhanshu; Swaminathan, Sankaran; Balasubramanian, Thangavel

2017-01-01

Thirty-one mercury-resistant bacterial strains were isolated from the effluent discharge sites of the SIPCOT industrial area. Among them, only one strain (CASKS5) was selected for further investigation due to its high minimum inhibitory concentration of mercury and low antibiotic susceptibility. In accordance with 16S ribosomal RNA gene sequences, the strain CASKS5 was identified as Vibrio fluvialis . The mercury-removal capacity of V. fluvialis was analyzed at four different concentrations (100, 150, 200, and 250  μ g/ml). Efficient bioremediation was observed at a level of 250  μ g/ml with the removal of 60% of mercury ions. The interesting outcome of this study was that the strain V. fluvialis had a high bioremediation efficiency but had a low antibiotic resistance. Hence, V. fluvialis could be successfully used as a strain for the ecofriendly removal of mercury.

Systems and methods for harvesting and storing materials produced in a nuclear reactor

DOEpatents

Heinold, Mark R.; Dayal, Yogeshwar; Brittingham, Martin W.

2016-04-05

Systems produce desired isotopes through irradiation in nuclear reactor instrumentation tubes and deposit the same in a robust facility for immediate shipping, handling, and/or consumption. Irradiation targets are inserted and removed through inaccessible areas without plant shutdown and placed in the harvesting facility, such as a plurality of sealable and shipping-safe casks and/or canisters. Systems may connect various structures in a sealed manner to avoid release of dangerous or unwanted matter throughout the nuclear plant, and/or systems may also automatically decontaminate materials to be released. Useable casks or canisters can include plural barriers for containment that are temporarily and selectively removable with specially-configured paths inserted therein. Penetrations in the facilities may limit waste or pneumatic gas escape and allow the same to be removed from the systems without over-pressurization or leakage. Methods include processing irradiation targets through such systems and securely delivering them in such harvesting facilities.

Role of meaningful subgroups in explaining differences in perceived variability for in-groups and out-groups.

PubMed

Park, B; Ryan, C S; Judd, C M

1992-10-01

Five aspects of the complexity of the knowledge representation of business and engineering majors were examined to see whether these differed by group membership and whether these differences were related to differences in perceived variability. Significantly more subgroups were generated when describing the in-group than the out-group; this difference predicted the relative tendency to see the in-group as more variable, and when controlled for statistically, out-group homogeneity effects were eliminated. Familiarity, redundancy, number of attributes used to describe the group, and the deviance of the subgroups from the larger group generally showed differences for in-group and out-group but did not show consistent evidence of mediation. In a 2nd study, Ss who were asked to sort group members into meaningful subgroups perceived greater variability relative to those who did not perform the sorting task.

VPS36-Mediated plasma membrane protein turnover is critical for Arabidopsis root gravitropism.

PubMed

Hsu, Ya-Wen; Jauh, Guang-Yuh

2017-04-03

The gravitropic response is an evolutionary adaptation for plants to cope with the altered gravitational field. It involves reestablishing the distribution of the phytohormone auxin by differential degradation of auxin influx and efflux carriers. This process includes the endosomal sorting complexes required for transport (ESCRT) machinery to recognize ubiquitinated proteins and deliver them to vacuoles for degradation, as evidenced by vps36-1 mutants. Here, we generated RNAi knockdown plants of Vacuolar Protein Sorting 36 (VPS36) that could survive to adulthood. VPS36-induced RNAi plants showed PIN FORMED1 (PIN1) accumulation in the intracellular compartment, reduced root length and small stature, as observed in vps36-1 mutants. After gravistimulation, the roots of VPS36-induced RNAi plants did not show the bending observed in wild-type plants. The VPS36-containing ESCRT machinery may have a role in the gravitropic response possibly associated with the degradation of auxin transporters.

Lipid phosphate phosphatase activity regulates dispersal and bilateral sorting of embryonic germ cells in Drosophila

PubMed Central

Renault, Andrew D.; Kunwar, Prabhat S.; Lehmann, Ruth

2010-01-01

In Drosophila, germ cell survival and directionality of migration are controlled by two lipid phosphate phosphatases (LPP), wunen (wun) and wunen-2 (wun2). wun wun2 double mutant analysis reveals that the two genes, hereafter collectively called wunens, act redundantly in primordial germ cells. We find that wunens mediate germ cell-germ cell repulsion and that this repulsion is necessary for germ cell dispersal and proper transepithelial migration at the onset of migration and for the equal sorting of the germ cells between the two embryonic gonads during their migration. We propose that this dispersal function optimizes adult fecundity by assuring maximal germ cell occupancy of both gonads. Furthermore, we find that the requirement for wunens in germ cell survival can be eliminated by blocking germ cell migration. We suggest that this essential function of Wunen is needed to maintain cell integrity in actively migrating germ cells. PMID:20431117

Management of the Cs/Sr Capsule Project at the Hanford Site. Technology Readiness Assessment Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

None, None

The Federal Project Director (FPD) for the U.S. Department of Energy (DOE), Richland Operations Office (RL) Waste Management and D&D Division (WMD) requested a Technology Readiness Assessment (TRA) for the Management of the Cesium/Strontium Capsule Storage Project (MCSCP) at the Waste Encapsulation and Storage Facility (WESF) on the Hanford Site in Washington State. The MCSCP CD-1 TRA was performed by a team selected in collaboration between the Office of Environmental Management (EM) Chief Engineer (EM-3.3) and RL, WMD FPD. The TRA Team included subject matter and technical experts having experience in cask storage, process engineering, and system design who weremore » independent of the MCSCP, and the team was led by the Director of Operations and Processes from the EM Chief Engineer's Office (EM-3.32). Movement of the Cs/Sr capsules to dry storage, based on information from the conceptual design, involves (1) capsule packaging, (2) capsule transfer, and (3) capsule storage. The project has developed a conceptual process, described in 30059-R-02, "NAC Conceptual Design Report for the Management of the Cesium and Strontium Capsules Project", which identifies the five major activities in the process to complete the transfer from storage pool to pad-mounted cask storage. The process, shown schematically in Figure 1, is comprised of the following process steps: (1) loading capsules into the UCS; (2) UCS processing; (3) UCS insertion into the TSC Basket; (4) cask transport from WESF to CSA and (5) extended storage at the CSA.« less

A FRAMEWORK TO DEVELOP FLAW ACCEPTANCE CRITERIA FOR STRUCTURAL INTEGRITY ASSESSMENT OF MULTIPURPOSE CANISTERS FOR EXTENDED STORAGE OF USED NUCLEAR FUEL

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lam, P.; Sindelar, R.; Duncan, A.

2014-04-07

A multipurpose canister (MPC) made of austenitic stainless steel is loaded with used nuclear fuel assemblies and is part of the transfer cask system to move the fuel from the spent fuel pool to prepare for storage, and is part of the storage cask system for on-site dry storage. This weld-sealed canister is also expected to be part of the transportation package following storage. The canister may be subject to service-induced degradation especially if exposed to aggressive environments during possible very long-term storage period if the permanent repository is yet to be identified and readied. Stress corrosion cracking may bemore » initiated on the canister surface in the welds or in the heat affected zone because the construction of MPC does not require heat treatment for stress relief. An acceptance criteria methodology is being developed for flaw disposition should the crack-like defects be detected by periodic Inservice Inspection. The external loading cases include thermal accident scenarios and cask drop conditions with the contribution from the welding residual stresses. The determination of acceptable flaw size is based on the procedure to evaluate flaw stability provided by American Petroleum Institute (API) 579 Fitness-for-Service (Second Edition). The material mechanical and fracture properties for base and weld metals and the stress analysis results are obtained from the open literature such as NUREG-1864. Subcritical crack growth from stress corrosion cracking (SCC), and its impact on inspection intervals and acceptance criteria, is not addressed.« less

10 CFR 72.214 - List of approved spent fuel storage casks.

Code of Federal Regulations, 2010 CFR

2010-01-01

... STORAGE OF SPENT NUCLEAR FUEL, HIGH-LEVEL RADIOACTIVE WASTE, AND REACTOR-RELATED GREATER THAN CLASS C WASTE General License for Storage of Spent Fuel at Power Reactor Sites § 72.214 List of approved spent...

10 CFR 72.214 - List of approved spent fuel storage casks.

Code of Federal Regulations, 2011 CFR

2011-01-01

... STORAGE OF SPENT NUCLEAR FUEL, HIGH-LEVEL RADIOACTIVE WASTE, AND REACTOR-RELATED GREATER THAN CLASS C WASTE General License for Storage of Spent Fuel at Power Reactor Sites § 72.214 List of approved spent...

76 FR 12825 - List of Approved Spent Fuel Storage Casks: NUHOMS® HD System Revision 1; Confirmation of...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-09

... definitions for Damaged Fuel Assembly and Transfer Operations; add definitions for Fuel Class and Reconstituted Fuel Assembly; add Combustion Engineering 16x16 class fuel assemblies as authorized contents...

Integrated System for Retrieval, Transportation and Consolidated Storage of Used Nuclear Fuel in the US - 13312

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bracey, William; Bondre, Jayant; Shelton, Catherine

2013-07-01

The current inventory of used nuclear fuel assemblies (UNFAs) from commercial reactor operations in the United States totals approximately 65,000 metric tons or approximately 232,000 UNFAs primarily stored at the 104 operational reactors in the US and a small number of decommissioned reactors. This inventory is growing at a rate of roughly 2,000 to 2,400 metric tons each year, (Approx. 7,000 UNFAs) as a result of ongoing commercial reactor operations. Assuming an average of 10 metric tons per storage/transportation casks, this inventory of commercial UNFAs represents about 6,500 casks with an additional of about 220 casks every year. In Januarymore » 2010, the Blue Ribbon Commission (BRC) [1] was directed to conduct a comprehensive review of policies for managing the back end of the nuclear fuel cycle and recommend a new plan. The BRC issued their final recommendations in January 2012. One of the main recommendations is for the United States to proceed promptly to develop one or more consolidated storage facilities (CSF) as part of an integrated, comprehensive plan for safely managing the back end of the nuclear fuel cycle. Based on its extensive experience in storage and transportation cask design, analysis, licensing, fabrication, and operations including transportation logistics, Transnuclear, Inc. (TN), an AREVA Subsidiary within the Logistics Business Unit, is engineering an integrated system that will address the complete process of commercial UNFA management. The system will deal with UNFAs in their current storage mode in various configurations, the preparation including handling and additional packaging where required and transportation of UNFAs to a CSF site, and subsequent storage, operation and maintenance at the CSF with eventual transportation to a future repository or recycling site. It is essential to proceed by steps to ensure that the system will be the most efficient and serve at best its purpose by defining: the problem to be resolved, the criteria to evaluate the solutions, and the alternative solutions. The complexity of the project is increasing with time (more fuel assemblies, new storage systems, deteriorating logistics infrastructure at some sites, etc.) but with the uncertainty on the final disposal path, flexibility and simplicity will be critical. (authors)« less

Coolant Density and Control Blade History Effects in Extended BWR Burnup Credit

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ade, Brian J; Marshall, William BJ J; Bowman, Stephen M

2015-01-01

Oak Ridge National Laboratory and the US Nuclear Regulatory Commission have initiated a multiyear project to investigate the application of burnup credit (BUC) for boiling water reactor (BWR) fuel in storage and transportation casks. This project includes two phases. The first phase investigates the applicability of peak reactivity methods currently used for spent fuel pools to spent fuel storage and transportation casks and the validation of reactivity (k eff) calculations and predicted spent fuel compositions. The second phase focuses on extending BUC beyond peak reactivity. This paper documents work performed to date investigating some aspects of extended BUC. (The technicalmore » basis for application of peak reactivity methods to BWR fuel in storage and transportation systems is presented in a companion paper.) Two reactor operating parameters are being evaluated to establish an adequate basis for extended BWR BUC: (1) the effect of axial void profile and (2) the effect of control blade utilization during operation. A detailed analysis of core simulator data for one cycle of a modern operating BWR plant was performed to determine the range of void profiles and the variability of the profile experienced during irradiation. Although a single cycle does not provide complete data, the data obtained are sufficient to determine the primary effects and to identify conservative modeling approaches. These data were used in a study of the effect of axial void profile. The first stage of the study was determination of the necessary moderator density temporal fidelity in depletion modeling. After the required temporal fidelity was established, multiple void profiles were used to examine the effect on cask reactivity. The results of these studies are being used to develop recommendations for conservatively modeling the void profile effects for BWR depletion calculations. The second operational parameter studied was control blade history. Control blades are inserted in various locations and at varying degrees during BWR operation based on the core loading pattern. When present during depletion, control blades harden the neutron spectrum locally because they displace the moderator and absorb thermal neutrons. The investigation of the effect of control blades on post operational cask reactivity is documented herein, as is the effect of multiple (continuous and intermittent) exposure periods with control blades inserted. The coupled effects of control blade presence on power density, void profile, or burnup profile will be addressed in future work.« less

Targeting of Drugs to ICAM for Treatment of Acute Lung Injury

DTIC Science & Technology

2005-04-01

cystein and aspartic immunoproteosome, and the hybrid proteosome), which proteases) [237, 239]. These proteases are extensively contain a common core...activation of the hybrid system, is clathrin-mediated internalization, although this proteosome (depending on ATP but not on ubiquitin) can also sort...21lbldIGNswsijceendoyti veicls . he cllswer thn wshe, inubaed t 3 0 C o a into a separate group to control for mechanical retention of second-dose

Studies of Altered Response to Infection Induced by Thermal Injury.

DTIC Science & Technology

1984-05-01

beginning to examine various prophylactic treatments for their efficacy in reversing a number of burn mediated immune defects. We expect even more interesting...with fluoresceinated antibody (OKM5) and FACS sorting or negative selection by Sephadex G-l0 passage and/or antibody and -3- complement treatment ... infectious episode. This mitogen hyperimmunity is typical of a normal immune system dealing with an infectious challenge. The Group II patients have a

Key mediators of intracellular amino acids signaling to mTORC1 activation.

PubMed

Duan, Yehui; Li, Fengna; Tan, Kunrong; Liu, Hongnan; Li, Yinghui; Liu, Yingying; Kong, Xiangfeng; Tang, Yulong; Wu, Guoyao; Yin, Yulong

2015-05-01

Mammalian target of rapamycin complex 1 (mTORC1) is activated by amino acids to promote cell growth via protein synthesis. Specifically, Ras-related guanosine triphosphatases (Rag GTPases) are activated by amino acids, and then translocate mTORC1 to the surface of late endosomes and lysosomes. Ras homolog enriched in brain (Rheb) resides on this surface and directly activates mTORC1. Apart from the presence of intracellular amino acids, Rag GTPases and Rheb, other mediators involved in intracellular amino acid signaling to mTORC1 activation include human vacuolar sorting protein-34 (hVps34) and mitogen-activating protein kinase kinase kinase kinase-3 (MAP4K3). Those molecular links between mTORC1 and its mediators form a complicate signaling network that controls cellular growth, proliferation, and metabolism. Moreover, it is speculated that amino acid signaling to mTORC1 may start from the lysosomal lumen. In this review, we discussed the function of these mediators in mTORC1 pathway and how these mediators are regulated by amino acids in details.

Peroxisomal membrane ascorbate peroxidase is sorted to a membranous network that resembles a subdomain of the endoplasmic reticulum.

PubMed Central

Mullen, R T; Lisenbee, C S; Miernyk, J A; Trelease, R N

1999-01-01

The peroxisomal isoform of ascorbate peroxidase (APX) is a novel membrane isoform that functions in the regeneration of NAD(+) and protection against toxic reactive oxygen species. The intracellular localization and sorting of peroxisomal APX were examined both in vivo and in vitro. Epitope-tagged peroxisomal APX, which was expressed transiently in tobacco BY-2 cells, localized to a reticular/circular network that resembled endoplasmic reticulum (ER; 3,3'-dihexyloxacarbocyanine iodide-stained membranes) and to peroxisomes. The reticular network did not colocalize with other organelle marker proteins, including three ER reticuloplasmins. However, in vitro, peroxisomal APX inserted post-translationally into the ER but not into other purified organelle membranes (including peroxisomal membranes). Insertion into the ER depended on the presence of molecular chaperones and ATP. These results suggest that regions of the ER serve as a possible intermediate in the sorting pathway of peroxisomal APX. Insight into this hypothesis was obtained from in vivo experiments with brefeldin A (BFA), a toxin that blocks vesicle-mediated protein export from ER. A transiently expressed chloramphenicol acetyltransferase-peroxisomal APX (CAT-pAPX) fusion protein accumulated only in the reticular/circular network in BFA-treated cells; after subsequent removal of BFA from these cells, the CAT-pAPX was distributed to preexisting peroxisomes. Thus, plant peroxisomal APX, a representative enzymatic peroxisomal membrane protein, is sorted to peroxisomes through an indirect pathway involving a preperoxisomal compartment with characteristics of a distinct subdomain of the ER, possibly a peroxisomal ER subdomain. PMID:10559442

A targeted boost-and-sort immunization strategy using Escherichia coli BamA identifies rare growth inhibitory antibodies.

PubMed

Vij, Rajesh; Lin, Zhonghua; Chiang, Nancy; Vernes, Jean-Michel; Storek, Kelly M; Park, Summer; Chan, Joyce; Meng, Y Gloria; Comps-Agrar, Laetitia; Luan, Peng; Lee, Sophia; Schneider, Kellen; Bevers, Jack; Zilberleyb, Inna; Tam, Christine; Koth, Christopher M; Xu, Min; Gill, Avinash; Auerbach, Marcy R; Smith, Peter A; Rutherford, Steven T; Nakamura, Gerald; Seshasayee, Dhaya; Payandeh, Jian; Koerber, James T

2018-05-08

Outer membrane proteins (OMPs) in Gram-negative bacteria are essential for a number of cellular functions including nutrient transport and drug efflux. Escherichia coli BamA is an essential component of the OMP β-barrel assembly machinery and a potential novel antibacterial target that has been proposed to undergo large (~15 Å) conformational changes. Here, we explored methods to isolate anti-BamA monoclonal antibodies (mAbs) that might alter the function of this OMP and ultimately lead to bacterial growth inhibition. We first optimized traditional immunization approaches but failed to identify mAbs that altered cell growth after screening >3000 hybridomas. We then developed a "targeted boost-and-sort" strategy that combines bacterial cell immunizations, purified BamA protein boosts, and single hybridoma cell sorting using amphipol-reconstituted BamA antigen. This unique workflow improves the discovery efficiency of FACS + mAbs by >600-fold and enabled the identification of rare anti-BamA mAbs with bacterial growth inhibitory activity in the presence of a truncated lipopolysaccharide layer. These mAbs represent novel tools for dissecting the BamA-mediated mechanism of β-barrel folding and our workflow establishes a new template for the efficient discovery of novel mAbs against other highly dynamic membrane proteins.

Sorting nexin 27 (SNX27) regulates the trafficking and activity of the glutamine transporter ASCT2.

PubMed

Yang, Zhe; Follett, Jordan; Kerr, Markus C; Clairfeuille, Thomas; Chandra, Mintu; Collins, Brett M; Teasdale, Rohan D

2018-05-04

Alanine-, serine-, cysteine-preferring transporter 2 (ASCT2, SLC1A5) is responsible for the uptake of glutamine into cells, a major source of cellular energy and a key regulator of mammalian target of rapamycin (mTOR) activation. Furthermore, ASCT2 expression has been reported in several human cancers, making it a potential target for both diagnostic and therapeutic purposes. Here we identify ASCT2 as a membrane-trafficked cargo molecule, sorted through a direct interaction with the PDZ domain of sorting nexin 27 (SNX27). Using both membrane fractionation and subcellular localization approaches, we demonstrate that the majority of ASCT2 resides at the plasma membrane. This is significantly reduced within CrispR-mediated SNX27 knockout (KO) cell lines, as it is missorted into the lysosomal degradation pathway. The reduction of ASCT2 levels in SNX27 KO cells leads to decreased glutamine uptake, which, in turn, inhibits cellular proliferation. SNX27 KO cells also present impaired activation of the mTOR complex 1 (mTORC1) pathway and enhanced autophagy. Taken together, our data reveal a role for SNX27 in glutamine uptake and amino acid-stimulated mTORC1 activation via modulation of ASCT2 intracellular trafficking. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Role of the Retromer Complex in Neurodegenerative Diseases

PubMed Central

Li, Chaosi; Shah, Syed Zahid Ali; Zhao, Deming; Yang, Lifeng

2016-01-01

The retromer complex is a protein complex that plays a central role in endosomal trafficking. Retromer dysfunction has been linked to a growing number of neurological disorders. The process of intracellular trafficking and recycling is crucial for maintaining normal intracellular homeostasis, which is partly achieved through the activity of the retromer complex. The retromer complex plays a primary role in sorting endosomal cargo back to the cell surface for reuse, to the trans-Golgi network (TGN), or alternatively to specialized endomembrane compartments, in which the cargo is not subjected to lysosomal-mediated degradation. In most cases, the retromer acts as a core that interacts with associated proteins, including sorting nexin family member 27 (SNX27), members of the vacuolar protein sorting 10 (VPS10) receptor family, the major endosomal actin polymerization-promoting complex known as Wiskott-Aldrich syndrome protein and scar homolog (WASH), and other proteins. Some of the molecules carried by the retromer complex are risk factors for neurodegenerative diseases. Defects such as haplo-insufficiency or mutations in one or several units of the retromer complex lead to various pathologies. Here, we summarize the molecular architecture of the retromer complex and the roles of this system in intracellular trafficking related the pathogenesis of neurodegenerative diseases. PMID:26973516

Delineating functional principles of the bow tie structure of a kinase-phosphatase network in the budding yeast.

PubMed

Abd-Rabbo, Diala; Michnick, Stephen W

2017-03-16

Kinases and phosphatases (KP) form complex self-regulating networks essential for cellular signal processing. In spite of having a wealth of data about interactions among KPs and their substrates, we have very limited models of the structures of the directed networks they form and consequently our ability to formulate hypotheses about how their structure determines the flow of information in these networks is restricted. We assembled and studied the largest bona fide kinase-phosphatase network (KP-Net) known to date for the yeast Saccharomyces cerevisiae. Application of the vertex sort (VS) algorithm on the KP-Net allowed us to elucidate its hierarchical structure in which nodes are sorted into top, core and bottom layers, forming a bow tie structure with a strongly connected core layer. Surprisingly, phosphatases tend to sort into the top layer, implying they are less regulated by phosphorylation than kinases. Superposition of the widest range of KP biological properties over the KP-Net hierarchy shows that core layer KPs: (i), receive the largest number of inputs; (ii), form bottlenecks implicated in multiple pathways and in decision-making; (iii), and are among the most regulated KPs both temporally and spatially. Moreover, top layer KPs are more abundant and less noisy than those in the bottom layer. Finally, we showed that the VS algorithm depends on node degrees without biasing the biological results of the sorted network. The VS algorithm is available as an R package ( https://cran.r-project.org/web/packages/VertexSort/index.html ). The KP-Net model we propose possesses a bow tie hierarchical structure in which the top layer appears to ensure highest fidelity and the core layer appears to mediate signal integration and cell state-dependent signal interpretation. Our model of the yeast KP-Net provides both functional insight into its organization as we understand today and a framework for future investigation of information processing in yeast and eukaryotes in general.

Protein Interacting with C Kinase 1 (PICK1) Reduces Reinsertion Rates of Interaction Partners Sorted to Rab11-dependent Slow Recycling Pathway*

PubMed Central

Madsen, Kenneth L.; Thorsen, Thor S.; Rahbek-Clemmensen, Troels; Eriksen, Jacob; Gether, Ulrik

2012-01-01

The scaffolding protein PICK1 (protein interacting with C kinase 1) contains an N-terminal PSD-95/Discs large/ZO-1 (PDZ) domain and a central lipid-binding Bin/amphiphysin/Rvs (BAR) domain. PICK1 is thought to regulate trafficking of its PDZ binding partners but different and even opposing functions have been suggested. Here, we apply ELISA-based assays and confocal microscopy in HEK293 cells with inducible PICK1 expression to assess in an isolated system the ability of PICK1 to regulate trafficking of natural and engineered PDZ binding partners. The dopamine transporter (DAT), which primarily sorts to degradation upon internalization, did not form perinuclear clusters with PICK1, and PICK1 did not affect DAT internalization/recycling. However, transfer of the PICK1-binding DAT C terminus to the β2-adrenergic receptor, which sorts to recycling upon internalization, led to formation of PICK1 co-clusters in Rab11-positive compartments. Furthermore, PICK1 inhibited Rab11-mediated recycling of the receptor in a BAR and PDZ domain-dependent manner. In contrast, transfer of the DAT C terminus to the δ-opioid receptor, which sorts to degradation, did not result in PICK1 co-clusters or any change in internalization/recycling. Further support for a role of PICK1 determined by its PDZ cargo was obtained for the PICK1 interaction partner prolactin-releasing peptide receptor (GPR10). GPR10 co-localized with Rab11 and clustered with PICK1 upon constitutive internalization but co-localized with the late endosomal marker Rab7 and did not cluster with PICK1 upon agonist-induced internalization. Our data suggest a selective role of PICK1 in clustering and reducing the recycling rates of PDZ domain binding partners sorted to the Rab11-dependent recycling pathway. PMID:22303009

Self-assembled Targeting of Cancer Cells by Iron(III)-doped, Silica Nanoparticles.

PubMed

Mitchell, K K Pohaku; Sandoval, S; Cortes-Mateos, M J; Alfaro, J G; Kummel, A C; Trogler, W C

2014-12-07

Iron(III)-doped silica nanoshells are shown to possess an in vitro cell-receptor mediated targeting functionality for endocytosis. Compared to plain silica nanoparticles, iron enriched ones are shown to be target-specific, a property that makes them potentially better vehicles for applications, such as drug delivery and tumor imaging, by making them more selective and thereby reducing the nanoparticle dose. Iron(III) in the nanoshells can interact with endogenous transferrin, a serum protein found in mammalian cell culture media, which subsequently promotes transport of the nanoshells into cells by the transferrin receptor-mediated endocytosis pathway. The enhanced uptake of the iron(III)-doped nanoshells relative to undoped silica nanoshells by a transferrin receptor-mediated pathway was established using fluorescence and confocal microscopy in an epithelial breast cancer cell line. This process was also confirmed using fluorescence activated cell sorting (FACS) measurements that show competitive blocking of nanoparticle uptake by added holo-transferrin.

RNA-Mediated Epigenetic Programming of Genome Rearrangements

PubMed Central

Nowacki, Mariusz; Shetty, Keerthi; Landweber, Laura F.

2012-01-01

RNA, normally thought of as a conduit in gene expression, has a novel mode of action in ciliated protozoa. Maternal RNA templates provide both an organizing guide for DNA rearrangements and a template that can transport somatic mutations to the next generation. This opportunity for RNA-mediated genome rearrangement and DNA repair is profound in the ciliate Oxytricha, which deletes 95% of its germline genome during development in a process that severely fragments its chromosomes and then sorts and reorders the hundreds of thousands of pieces remaining. Oxytricha’s somatic nuclear genome is therefore an epigenome formed through RNA templates and signals arising from the previous generation. Furthermore, this mechanism of RNA-mediated epigenetic inheritance can function across multiple generations, and the discovery of maternal template RNA molecules has revealed new biological roles for RNA and has hinted at the power of RNA molecules to sculpt genomic information in cells. PMID:21801022

AP-1/σ1B-adaptin mediates endosomal synaptic vesicle recycling, learning and memory

PubMed Central

Glyvuk, Nataliya; Tsytsyura, Yaroslav; Geumann, Constanze; D'Hooge, Rudi; Hüve, Jana; Kratzke, Manuel; Baltes, Jennifer; Böning, Daniel; Klingauf, Jürgen; Schu, Peter

2010-01-01

Synaptic vesicle recycling involves AP-2/clathrin-mediated endocytosis, but it is not known whether the endosomal pathway is also required. Mice deficient in the tissue-specific AP-1–σ1B complex have impaired synaptic vesicle recycling in hippocampal synapses. The ubiquitously expressed AP-1–σ1A complex mediates protein sorting between the trans-Golgi network and early endosomes. Vertebrates express three σ1 subunit isoforms: A, B and C. The expressions of σ1A and σ1B are highest in the brain. Synaptic vesicle reformation in cultured neurons from σ1B-deficient mice is reduced upon stimulation, and large endosomal intermediates accumulate. The σ1B-deficient mice have reduced motor coordination and severely impaired long-term spatial memory. These data reveal a molecular mechanism for a severe human X-chromosome-linked mental retardation. PMID:20203623

324 Building spent fuel segments pieces and fragments removal summary report

DOE Office of Scientific and Technical Information (OSTI.GOV)

SMITH, C L

2003-01-09

As part of the 324 Building Deactivation Project, all Spent Nuclear Fuel (SNF) and Special Nuclear Material were removed. The removal entailed packaging the material into a GNS-12 cask and shipping it to the Central Waste Complex (CWC).

A&M. Radioactive parts security storage area. camera facing northwest. Outdoor ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

A&M. Radioactive parts security storage area. camera facing northwest. Outdoor storage of concrete storage casks. Photographer: M. Holmes. Date: November 21, 1959. INEEL negative no. 59-6081 - Idaho National Engineering Laboratory, Test Area North, Scoville, Butte County, ID

U.S. regulatory research program for implementation of burnup credit in transport casks

DOT National Transportation Integrated Search

2001-09-10

In 1999 the U.S. Nuclear Regulatory Commission (U.S. NRC) initiated a research program to : support the development of technical bases and guidance that would facilitate the implementation of : burnup credit into licensing activities for transport an...

10 CFR Appendix D to Part 73 - Physical Protection of Irradiated Reactor Fuel in Transit, Training Program Subject Schedule

Code of Federal Regulations, 2013 CFR

2013-01-01

... radioactive cargo —Function and characteristics of the shipping casks —Radiation hazards —Federal, State and... Contingencies —Accidents —Severe weather conditions —Vehicle breakdown —Communications problems —Radioactive...

10 CFR Appendix D to Part 73 - Physical Protection of Irradiated Reactor Fuel in Transit, Training Program Subject Schedule

Code of Federal Regulations, 2014 CFR

2014-01-01

... radioactive cargo —Function and characteristics of the shipping casks —Radiation hazards —Federal, State and... Contingencies —Accidents —Severe weather conditions —Vehicle breakdown —Communications problems —Radioactive...

10 CFR Appendix D to Part 73 - Physical Protection of Irradiated Reactor Fuel in Transit, Training Program Subject Schedule

Code of Federal Regulations, 2012 CFR

2012-01-01

... radioactive cargo —Function and characteristics of the shipping casks —Radiation hazards —Federal, State and... Contingencies —Accidents —Severe weather conditions —Vehicle breakdown —Communications problems —Radioactive...

Neutrophil degranulation and immunosuppression in patients with GBM: restoration of cellular immune function by targeting arginase I.

PubMed

Sippel, Trisha R; White, Jason; Nag, Kamalika; Tsvankin, Vadim; Klaassen, Marci; Kleinschmidt-DeMasters, B K; Waziri, Allen

2011-11-15

The source of glioblastoma (GBM)-associated immunosuppression remains multifactorial. We sought to clarify and therapeutically target myeloid cell-derived peripheral immunosuppression in patients with GBM. Direct ex vivo T-cell function, serum Arginase I (ArgI) levels, and circulating myeloid lineage populations were compared between patients with GBM and normal donors or patients with other intracranial tumors. Immunofunctional assays were conducted using bulk and sorted cell populations to explore the potential transfer of myeloid cell-mediated immunosuppression and to identify a potential mechanism for these effects. ArgI-mediated immunosuppression was therapeutically targeted in vitro through pharmacologic inhibition or arginine supplementation. We identified a significantly expanded population of circulating, degranulated neutrophils associated with elevated levels of serum ArgI and decreased T-cell CD3ζ expression within peripheral blood from patients with GBM. Sorted CD11b(+) cells from patients with GBM were found to markedly suppress normal donor T-cell function in coculture, and media harvested from mitogen-stimulated GBM peripheral blood mononuclear cell (PBMC) or GBM-associated mixed lymphoid reactions showed ArgI levels that were significantly higher than controls. Critically, T-cell suppression in both settings could be completely reversed through pharmacologic ArgI inhibition or with arginine supplementation. These data indicate that peripheral cellular immunosuppression in patients with GBM is associated with neutrophil degranulation and elevated levels of circulating ArgI, and that T-cell function can be restored in these individuals by targeting ArgI. These data identify a novel pathway of GBM-mediated suppression of cellular immunity and offer a potential therapeutic window for improving antitumor immunity in affected patients.

Discovery of Prostate Cancer Tumor Suppressors and Mediators of MDV3100 Resistance Through in Vivo RNA Interference Screen

DTIC Science & Technology

2015-11-01

concept of increase of NSE and decrease of PSMA with NED. Right, the quadrants of a hypothetical FACS-plot after Enzalutamide treatment from which...antigen ( PSMA ) expression. Thus, we also decided to sort for cells that have decreased PSMA expression with prolonged Enzalutamide treatment. Using the...cells, till there was almost plateauing of the GFP signals, Figure 6. PSMA expression FACS%based+screen+strategy+ AR#independent# AR

The MARVEL transmembrane motif of occludin mediates oligomerization and targeting to the basolateral surface in epithelia.

PubMed

Yaffe, Yakey; Shepshelovitch, Jeanne; Nevo-Yassaf, Inbar; Yeheskel, Adva; Shmerling, Hedva; Kwiatek, Joanna M; Gaus, Katharina; Pasmanik-Chor, Metsada; Hirschberg, Koret

2012-08-01

Occludin (Ocln), a MARVEL-motif-containing protein, is found in all tight junctions. MARVEL motifs are comprised of four transmembrane helices associated with the localization to or formation of diverse membrane subdomains by interacting with the proximal lipid environment. The functions of the Ocln MARVEL motif are unknown. Bioinformatics sequence- and structure-based analyses demonstrated that the MARVEL domain of Ocln family proteins has distinct evolutionarily conserved sequence features that are consistent with its basolateral membrane localization. Live-cell microscopy, fluorescence resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC) were used to analyze the intracellular distribution and self-association of fluorescent-protein-tagged full-length human Ocln or the Ocln MARVEL motif excluding the cytosolic C- and N-termini (amino acids 60-269, FP-MARVEL-Ocln). FP-MARVEL-Ocln efficiently arrived at the plasma membrane (PM) and was sorted to the basolateral PM in filter-grown polarized MDCK cells. A series of conserved aromatic amino acids within the MARVEL domain were found to be associated with Ocln dimerization using BiFC. FP-MARVEL-Ocln inhibited membrane pore growth during Triton-X-100-induced solubilization and was shown to increase the membrane-ordered state using Laurdan, a lipid dye. These data demonstrate that the Ocln MARVEL domain mediates self-association and correct sorting to the basolateral membrane.

Transposon-mediated generation of BCR-ABL1-expressing transgenic cell lines for unbiased sensitivity testing of tyrosine kinase inhibitors.

PubMed

Byrgazov, Konstantin; Lucini, Chantal Blanche; Berkowitsch, Bettina; Koenig, Margit; Haas, Oskar A; Hoermann, Gregor; Valent, Peter; Lion, Thomas

2016-11-22

Point mutations in the ABL1 kinase domain are an important mechanism of resistance to tyrosine kinase inhibitors (TKI) in BCR-ABL1-positive and, as recently shown, BCR-ABL1-like leukemias. The cell line Ba/F3 lentivirally transduced with mutant BCR-ABL1 constructs is widely used for in vitro sensitivity testing and response prediction to tyrosine kinase inhibitors. The transposon-based Sleeping Beauty system presented offers several advantages over lentiviral transduction including the absence of biosafety issues, faster generation of transgenic cell lines, and greater efficacy in introducing large gene constructs. Nevertheless, both methods can mediate multiple insertions in the genome. Here we show that multiple BCR-ABL1 insertions result in elevated IC50 levels for individual TKIs, thus overestimating the actual resistance of mutant subclones. We have therefore established flow-sorting-based fractionation of BCR-ABL1-transformed Ba/F3 cells facilitating efficient enrichment of cells carrying single-site insertions, as demonstrated by FISH-analysis. Fractions of unselected Ba/F3 cells not only showed a greater number of BCR-ABL1 hybridization signals, but also revealed higher IC50 values for the TKIs tested. The data presented highlight the need to carefully select transfected cells by flow-sorting, and to control the insertion numbers by FISH and real-time PCR to permit unbiased in vitro testing of drug resistance.

Distinguishing between incomplete lineage sorting and genomic introgressions: complete fixation of allospecific mitochondrial DNA in a sexually reproducing fish (Cobitis; Teleostei), despite clonal reproduction of hybrids.

PubMed

Choleva, Lukas; Musilova, Zuzana; Kohoutova-Sediva, Alena; Paces, Jan; Rab, Petr; Janko, Karel

2014-01-01

Distinguishing between hybrid introgression and incomplete lineage sorting causing incongruence among gene trees in that they exhibit topological differences requires application of statistical approaches that are based on biologically relevant models. Such study is especially challenging in hybrid systems, where usual vectors mediating interspecific gene transfers--hybrids with Mendelian heredity--are absent or unknown. Here we study a complex of hybridizing species, which are known to produce clonal hybrids, to discover how one of the species, Cobitis tanaitica, has achieved a pattern of mito-nuclear mosaic genome over the whole geographic range. We appplied three distinct methods, including the method using solely the information on gene tree topologies, and found that the contrasting mito-nuclear signal might not have resulted from the retention of ancestral polymorphism. Instead, we found two signs of hybridization events related to C. tanaitica; one concerning nuclear gene flow and the other suggested mitochondrial capture. Interestingly, clonal inheritance (gynogenesis) of contemporary hybrids prevents genomic introgressions and non-clonal hybrids are either absent or too rare to be detected among European Cobitis. Our analyses therefore suggest that introgressive hybridizations are rather old episodes, mediated by previously existing hybrids whose inheritance was not entirely clonal. Cobitis complex thus supports the view that the type of resulting hybrids depends on a level of genomic divergence between sexual species.

Identification of genes mediating thyroid hormone action in the developing mouse cerebellum.

PubMed

Takahashi, Masaki; Negishi, Takayuki; Tashiro, Tomoko

2008-02-01

Despite the indispensable role thyroid hormone (TH) plays in brain development, only a small number of genes have been identified to be directly regulated by TH and its precise mechanism of action remains largely unknown, partly because most of the previous studies have been carried out at postnatal day 15 or later. In the present study, we screened for TH-responsive genes in the developing mouse cerebellum at postnatal day 4 when morphological alterations because of TH status are not apparent. Among the new candidate genes selected by comparing gene expression profiles of experimentally hypothyroid, hypothyroid with postnatal thyroxine replacement, and control animals using oligoDNA microarrays, six genes were confirmed by real-time PCR to be positively (orc1l, galr3, sort1, nlgn3, cdk5r2, and zfp367) regulated by TH. Among these, sort1, cdk5r2, and zfp367 were up-regulated already at 1 h after a single injection of thyroxine to the hypothyroid or control animal, suggesting them to be possible primary targets of the hormone. Cell proliferation and apoptosis examined by BrdU incorporation and terminal deoxynucleotide transferase-mediated dUTP nick-end labeling assay revealed that hypothyroidism by itself did not enhance apoptosis at this stage, but rather increased cell survival, possibly through regulation of these newly identified genes.

Protein Mobilization in Germinating Mung Bean Seeds Involves Vacuolar Sorting Receptors and Multivesicular Bodies1[W][OA

PubMed Central

Wang, Junqi; Li, Yubing; Lo, Sze Wan; Hillmer, Stefan; Sun, Samuel S.M.; Robinson, David G.; Jiang, Liwen

2007-01-01

Plants accumulate and store proteins in protein storage vacuoles (PSVs) during seed development and maturation. Upon seed germination, these storage proteins are mobilized to provide nutrients for seedling growth. However, little is known about the molecular mechanisms of protein degradation during seed germination. Here we test the hypothesis that vacuolar sorting receptor (VSR) proteins play a role in mediating protein degradation in germinating seeds. We demonstrate that both VSR proteins and hydrolytic enzymes are synthesized de novo during mung bean (Vigna radiata) seed germination. Immunogold electron microscopy with VSR antibodies demonstrate that VSRs mainly locate to the peripheral membrane of multivesicular bodies (MVBs), presumably as recycling receptors in day 1 germinating seeds, but become internalized to the MVB lumen, presumably for degradation at day 3 germination. Chemical cross-linking and immunoprecipitation with VSR antibodies have identified the cysteine protease aleurain as a specific VSR-interacting protein in germinating seeds. Further confocal immunofluorescence and immunogold electron microscopy studies demonstrate that VSR and aleurain colocalize to MVBs as well as PSVs in germinating seeds. Thus, MVBs in germinating seeds exercise dual functions: as a storage compartment for proteases that are physically separated from PSVs in the mature seed and as an intermediate compartment for VSR-mediated delivery of proteases from the Golgi apparatus to the PSV for protein degradation during seed germination. PMID:17322331

ESCRT-II controls retinal axon growth by regulating DCC receptor levels and local protein synthesis

PubMed Central

Konopacki, Filip A.; Dwivedy, Asha; Bellon, Anaïs; Blower, Michael D.

2016-01-01

Endocytosis and local protein synthesis (LPS) act coordinately to mediate the chemotropic responses of axons, but the link between these two processes is poorly understood. The endosomal sorting complex required for transport (ESCRT) is a key regulator of cargo sorting in the endocytic pathway, and here we have investigated the role of ESCRT-II, a critical ESCRT component, in Xenopus retinal ganglion cell (RGC) axons. We show that ESCRT-II is present in RGC axonal growth cones (GCs) where it co-localizes with endocytic vesicle GTPases and, unexpectedly, with the Netrin-1 receptor, deleted in colorectal cancer (DCC). ESCRT-II knockdown (KD) decreases endocytosis and, strikingly, reduces DCC in GCs and leads to axon growth and guidance defects. ESCRT-II-depleted axons fail to turn in response to a Netrin-1 gradient in vitro and many axons fail to exit the eye in vivo. These defects, similar to Netrin-1/DCC loss-of-function phenotypes, can be rescued in whole (in vitro) or in part (in vivo) by expressing DCC. In addition, ESCRT-II KD impairs LPS in GCs and live imaging reveals that ESCRT-II transports mRNAs in axons. Collectively, our results show that the ESCRT-II-mediated endocytic pathway regulates both DCC and LPS in the axonal compartment and suggest that ESCRT-II aids gradient sensing in GCs by coupling endocytosis to LPS. PMID:27248654

SLUDGE TREATMENT PROJECT COST COMPARISON BETWEEN HYDRAULIC LOADING AND SMALL CANISTER LOADING CONCEPTS

DOE Office of Scientific and Technical Information (OSTI.GOV)

GEUTHER J; CONRAD EA; RHOADARMER D

2009-08-24

The Sludge Treatment Project (STP) is considering two different concepts for the retrieval, loading, transport and interim storage of the K Basin sludge. The two design concepts under consideration are: (1) Hydraulic Loading Concept - In the hydraulic loading concept, the sludge is retrieved from the Engineered Containers directly into the Sludge Transport and Storage Container (STSC) while located in the STS cask in the modified KW Basin Annex. The sludge is loaded via a series of transfer, settle, decant, and filtration return steps until the STSC sludge transportation limits are met. The STSC is then transported to T Plantmore » and placed in storage arrays in the T Plant canyon cells for interim storage. (2) Small Canister Concept - In the small canister concept, the sludge is transferred from the Engineered Containers (ECs) into a settling vessel. After settling and decanting, the sludge is loaded underwater into small canisters. The small canisters are then transferred to the existing Fuel Transport System (FTS) where they are loaded underwater into the FTS Shielded Transfer Cask (STC). The STC is raised from the basin and placed into the Cask Transfer Overpack (CTO), loaded onto the trailer in the KW Basin Annex for transport to T Plant. At T Plant, the CTO is removed from the transport trailer and placed on the canyon deck. The CTO and STC are opened and the small canisters are removed using the canyon crane and placed into an STSC. The STSC is closed, and placed in storage arrays in the T Plant canyon cells for interim storage. The purpose of the cost estimate is to provide a comparison of the two concepts described.« less

AIR SHIPMENT OF HIGHLY ENRICHED URANIUM SPENT NUCLEAR FUEL FROM ROMANIA AND LIBYA

DOE Office of Scientific and Technical Information (OSTI.GOV)

Christopher Landers; Igor Bolshinsky; Ken Allen

2010-07-01

In June 2009 Romania successfully completed the world’s first air shipment of highly enriched uranium (HEU) spent nuclear fuel transported in Type B(U) casks under existing international laws and without special exceptions for the air transport licenses. Special 20-foot ISO shipping containers and cask tiedown supports were designed to transport Russian TUK 19 shipping casks for the Romanian air shipment and the equipment was certified for all modes of transport, including road, rail, water, and air. In December 2009 Libya successfully used this same equipment for a second air shipment of HEU spent nuclear fuel. Both spent fuel shipments weremore » transported by truck from the originating nuclear facilities to nearby commercial airports, were flown by commercial cargo aircraft to a commercial airport in Yekaterinburg, Russia, and then transported by truck to their final destinations at the Production Association Mayak facility in Chelyabinsk, Russia. Both air shipments were performed under the Russian Research Reactor Fuel Return Program (RRRFR) as part of the U.S. National Nuclear Security Administration (NNSA) Global Threat Reduction Initiative (GTRI). The Romania air shipment of 23.7 kg of HEU spent fuel from the VVR S research reactor was the last of three HEU fresh and spent fuel shipments under RRRFR that resulted in Romania becoming the 3rd RRRFR participating country to remove all HEU. Libya had previously completed two RRRFR shipments of HEU fresh fuel so the 5.2 kg of HEU spent fuel air shipped from the IRT 1 research reactor in December made Libya the 4th RRRFR participating country to remove all HEU. This paper describes the equipment, preparations, and license approvals required to safely and securely complete these two air shipments of spent nuclear fuel.« less

Air Shipment of Spent Nuclear Fuel from Romania to Russia

DOE Office of Scientific and Technical Information (OSTI.GOV)

Igor Bolshinsky; Ken Allen; Lucian Biro

Romania successfully completed the world’s first air shipment of spent nuclear fuel transported in Type B(U) casks under existing international laws and without shipment license special exceptions when the last Romanian highly enriched uranium (HEU) spent nuclear fuel was transported to the Russian Federation in June 2009. This air shipment required the design, fabrication, and licensing of special 20 foot freight containers and cask tiedown supports to transport the eighteen TUK 19 shipping casks on a Russian commercial cargo aircraft. The new equipment was certified for transport by road, rail, water, and air to provide multi modal transport capabilities formore » shipping research reactor spent fuel. The equipment design, safety analyses, and fabrication were performed in the Russian Federation and transport licenses were issued by both the Russian and Romanian regulatory authorities. The spent fuel was transported by truck from the VVR S research reactor to the Bucharest airport, flown by commercial cargo aircraft to the airport at Yekaterinburg, Russia, and then transported by truck to the final destination in a secure nuclear facility at Chelyabinsk, Russia. This shipment of 23.7 kg of HEU was coordinated by the Russian Research Reactor Fuel Return Program (RRRFR), as part of the U.S. Department of Energy Global Threat Reduction Initiative (GTRI), in close cooperation with the Rosatom State Atomic Energy Corporation and the International Atomic Energy Agency, and was managed in Romania by the National Commission for Nuclear Activities Control (CNCAN). This paper describes the planning, shipment preparations, equipment design, and license approvals that resulted in the safe and secure air shipment of this spent nuclear fuel.« less

Alcohol promotions in Australian supermarket catalogues.

PubMed

Johnston, Robyn; Stafford, Julia; Pierce, Hannah; Daube, Mike

2017-07-01

In Australia, most alcohol is sold as packaged liquor from off-premises retailers, a market increasingly dominated by supermarket chains. Competition between retailers may encourage marketing approaches, for example, discounting, that evidence indicates contribute to alcohol-related harms. This research documented the nature and variety of promotional methods used by two major supermarket retailers to promote alcohol products in their supermarket catalogues. Weekly catalogues from the two largest Australian supermarket chains were reviewed for alcohol-related content over 12 months. Alcohol promotions were assessed for promotion type, product type, number of standard drinks, purchase price and price/standard drink. Each store catalogue included, on average, 13 alcohol promotions/week, with price-based promotions most common. Forty-five percent of promotions required the purchase of multiple alcohol items. Wine was the most frequently promoted product (44%), followed by beer (24%) and spirits (18%). Most (99%) wine cask (2-5 L container) promotions required multiple (two to three) casks to be purchased. The average number of standard drinks required to be purchased to participate in catalogue promotions was 31.7 (SD = 24.9; median = 23.1). The median price per standard drink was $1.49 (range $0.19-$9.81). Cask wines had the lowest cost per standard drink across all product types. Supermarket catalogues' emphasis on low prices/high volumes of alcohol reflects that retailers are taking advantage of limited restrictions on off-premise sales and promotion, which allow them to approach market competition in ways that may increase alcohol-related harms in consumers. Regulation of alcohol marketing should address retailer catalogue promotions. [Johnston R, Stafford J, Pierce H, Daube M. Alcohol promotions in Australian supermarket catalogues. Drug Alcohol Rev 2017;36:456-463]. © 2016 Australasian Professional Society on Alcohol and other Drugs.

Environmental data and analyses for the proposed management of spent nuclear fuel on the DOE Oak Ridge Reservation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Socolof, M.L.; Curtis, A.H.; Blasing, T.J.

1995-08-01

DOE needs to continue the safe and efficient management of SNF on ORR, based on the requirement for future SNF storage capacity and implementation of the ROD for the PEIS. DOE is proposing to implement the ROD through proper management of SNF on ORR, including the possible construction and operation of a dry cask storage facility. This report describes the potentially affected environment and analyzes impacts on various resources due to the proposed action. The information provided in this report is intended to support the Environmental Assessment being prepared for the proposed activities. Construction of the dry cask storage facilitymore » would result in minimal or no impacts on groundwater, surface water, and ecological resources. Contaminated soils excavated during construction would result in negligible risk to human health and to biota. Except for noise from trucks and equipment, operation of the dry cask storage facility would not be expected to have any impact on vegetation, wildlife, or rare plants or animals. Noise impacts would be minimal. Operation exposures to the average SNF storage facility worker would not exceed approximately 0.40 mSv/year (40 mrem/year). The off-site population dose within an 80-km (50-mile) radius of ORR from SNF operations would be less than 0.052 person-Sv/year (5.2 person-rem/year). Impacts from incident-free transportation on ORR would be less than 1.36 X 10{sup -4} occupational fatal cancers and 4.28 X 10{sup -6} public fatal cancers. Credible accident scenarios that would result in the greatest probable risks would cause less than one in a million cancer fatalities to workers and the public.« less

Numerical Estimation of the Spent Fuel Ratio

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lindgren, Eric R.; Durbin, Samuel; Wilke, Jason

Sabotage of spent nuclear fuel casks remains a concern nearly forty years after attacks against shipment casks were first analyzed and has a renewed relevance in the post-9/11 environment. A limited number of full-scale tests and supporting efforts using surrogate materials, typically depleted uranium dioxide (DUO 2 ), have been conducted in the interim to more definitively determine the source term from these postulated events. However, the validity of these large- scale results remain in question due to the lack of a defensible spent fuel ratio (SFR), defined as the amount of respirable aerosol generated by an attack on amore » mass of spent fuel compared to that of an otherwise identical surrogate. Previous attempts to define the SFR in the 1980's have resulted in estimates ranging from 0.42 to 12 and include suboptimal experimental techniques and data comparisons. Because of the large uncertainty surrounding the SFR, estimates of releases from security-related events may be unnecessarily conservative. Credible arguments exist that the SFR does not exceed a value of unity. A defensible determination of the SFR in this lower range would greatly reduce the calculated risk associated with the transport and storage of spent nuclear fuel in dry cask systems. In the present work, the shock physics codes CTH and ALE3D were used to simulate spent nuclear fuel (SNF) and DUO 2 targets impacted by a high-velocity jet at an ambient temperature condition. These preliminary results are used to illustrate an approach to estimate the respirable release fraction for each type of material and ultimately, an estimate of the SFR. This page intentionally blank« less

27 CFR 28.211 - General.

Code of Federal Regulations, 2010 CFR

2010-04-01

... TREASURY LIQUORS EXPORTATION OF ALCOHOL Exportation of Wine With Benefit of Drawback § 28.211 General. Wines manufactured, produced, bottled in bottles packed in containers, or packaged in casks or other... which are filled on premises qualified under this chapter to package or bottle wines, may, subject to...

Radionuclide production and dose rate estimation during the commissioning of the W-Ta spallation target

NASA Astrophysics Data System (ADS)

Yu, Q. Z.; Liang, T. J.

2018-06-01

China Spallation Neutron Source (CSNS) is intended to begin operation in 2018. CSNS is an accelerator-base multidisciplinary user facility. The pulsed neutrons are produced by a 1.6GeV short-pulsed proton beam impinging on a W-Ta spallation target, at a beam power of100 kW and a repetition rate of 25 Hz. 20 neutron beam lines are extracted for the neutron scattering and neutron irradiation research. During the commissioning and maintenance scenarios, the gamma rays induced from the W-Ta target can cause the dose threat to the personal and the environment. In this paper, the gamma dose rate distributions for the W-Ta spallation are calculated, based on the engineering model of the target-moderator-reflector system. The shipping cask is analyzed to satisfy the dose rate limit that less than 2 mSv/h at the surface of the shipping cask. All calculations are performed by the Monte carlo code MCNPX2.5 and the activation code CINDER’90.

Draft evaluation of the frequency for gas sampling for the high burnup confirmatory data project

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stockman, Christine T.; Alsaed, Halim A.; Bryan, Charles R.

2015-03-26

This report fulfills the M3 milestone M3FT-15SN0802041, “Draft Evaluation of the Frequency for Gas Sampling for the High Burn-up Storage Demonstration Project” under Work Package FT-15SN080204, “ST Field Demonstration Support – SNL”. This report provides a technically based gas sampling frequency strategy for the High Burnup (HBU) Confirmatory Data Project. The evaluation of: 1) the types and magnitudes of gases that could be present in the project cask and, 2) the degradation mechanisms that could change gas compositions culminates in an adaptive gas sampling frequency strategy. This adaptive strategy is compared against the sampling frequency that has been developed basedmore » on operational considerations. Gas sampling will provide information on the presence of residual water (and byproducts associated with its reactions and decomposition) and breach of cladding, which could inform the decision of when to open the project cask.« less

Usage of FT-ICR-MS Metabolomics for Characterizing the Chemical Signatures of Barrel-Aged Whisky

PubMed Central

Roullier-Gall, Chloé; Signoret, Julie; Hemmler, Daniel; Witting, Michael A.; Kanawati, Basem; Schäfer, Bernhard; Gougeon, Régis D.; Schmitt-Kopplin, Philippe

2018-01-01

Whisky can be described as a complex matrix integrating the chemical history from the fermented cereals, the wooden barrels, the specific distillery processes, aging, and environmental factors. In this study, using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), we analyzed 150 whisky samples from 49 different distilleries, 7 countries, and ranging from 1 day new make spirit to 43 years of maturation with different types of barrel. Chemometrics revealed the unexpected impact of the wood history on the distillate's composition during barrel aging, regardless of the whisky origin. Flavonols, oligolignols, and fatty acids are examples of important chemical signatures for Bourbon casks, whereas a high number of polyphenol glycosides, including for instance quercetin-glucuronide or myricetin-glucoside as potential candidates, and carbohydrates would discriminate Sherry casks. However, the comparison of barrel aged rums and whiskies revealed specific signatures, highlighting the importance of the initial composition of the distillate and the distillery processes. PMID:29520358

A methodology to quantify the release of spent nuclear fuel from dry casks during security-related scenarios

DOE Office of Scientific and Technical Information (OSTI.GOV)

Durbin, Samuel G.; Luna, Robert Earl

Assessing the risk to the public and the environment from a release of radioactive material produced by accidental or purposeful forces/environments is an important aspect of the regulatory process in many facets of the nuclear industry. In particular, the transport and storage of radioactive materials is of particular concern to the public, especially with regard to potential sabotage acts that might be undertaken by terror groups to cause injuries, panic, and/or economic consequences to a nation. For many such postulated attacks, no breach in the robust cask or storage module containment is expected to occur. However, there exists evidence thatmore » some hypothetical attack modes can penetrate and cause a release of radioactive material. This report is intended as an unclassified overview of the methodology for release estimation as well as a guide to useful resource data from unclassified sources and relevant analysis methods for the estimation process.« less

AIR SHIPMENT OF SPENT NUCLEAR FUEL FROM THE BUDAPEST RESEARCH REACTOR

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dewes, J.

2014-02-24

The shipment of spent nuclear fuel is usually done by a combination of rail, road or sea, as the high activity of the SNF needs heavy shielding. Air shipment has advantages, e.g. it is much faster than any other shipment and therefore minimizes the transit time as well as attention of the public. Up to now only very few and very special SNF shipments were done by air, as the available container (TUK6) had a very limited capacity. Recently Sosny developed a Type C overpack, the TUK-145/C, compliant with IAEA Standard TS-R-1 for the VPVR/M type Skoda container. The TUK-145/Cmore » was first used in Vietnam in July 2013 for a single cask. In October and November 2013 a total of six casks were successfully shipped from Hungary in three air shipments using the TUK-145/C. The present paper describes the details of these shipments and formulates the lessons learned.« less

Usage of FT-ICR-MS Metabolomics for characterizing the chemical signatures of barrel-aged whisky

NASA Astrophysics Data System (ADS)

Roullier-Gall, Chloé; Signoret, Julie; Hemmler, Daniel; Witting, Michael A.; Kanawati, Basem; Schäfer, Bernhard; Gougeon, Régis D.; Schmitt-Kopplin, Philippe

2018-02-01

Whisky can be described as a complex matrix integrating the chemical history from the fermented cereals, the wooden barrels, the specific distillery processes, ageing and environmental factors. In this study, using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), we analysed 150 whisky samples from 49 different distilleries, 7 countries, and ranging from 1 day new make spirit to 43 years of maturation with different types of barrel. Chemometrics revealed the unexpected impact of the wood history on the distillatés composition during barrel ageing, regardless of the whisky origin. Flavonols, oligolignols and fatty acids are examples of important chemical signatures for Bourbon casks, whereas a high number of polyphenol glycosides, including for instance quercetin-glucuronide or myricetin-glucoside as potential candidates, and carbohydrates would discriminate Sherry casks. However, the comparison of barrel aged rums and whiskies revealed specific signatures, highlighting the importance of the initial composition of the distillate and the distillery processes.

Apollo 12 Mission image - Modular Equipment Stowage Assemble (MESA) and the Fuel Cask on the Lunar Module (LM)

NASA Image and Video Library

1969-11-19

AS12-48-7034 (19 Nov. 1969) --- A close-up view of a portion of quadrant II of the descent stage of the Apollo 12 Lunar Module (LM), photographed during the Apollo 12 extravehicular activity (EVA). At lower left is the LM's Y footpad. The empty Radioisotope Thermoelectric Generator (RTG) fuel cask is at upper right. The fuel capsule has already been removed and placed in the RTG. The RTG furnishes power for the Apollo Lunar Surface Experiments Package (ALSEP) which the Apollo 12 astronauts deployed on the moon. The LM's descent engine skirt is in the center background. The rod-like object protruding out from under the footpad is a lunar surface sensing probe. Astronaut Richard F. Gordon Jr., command module pilot, remained with the Command and Service Modules (CSM) in lunar orbit while astronauts Charles Conrad Jr., commander; and Alan L. Bean, lunar module pilot, descended in the LM to explore the moon.

Behavioral vs. molecular sources of conflict between nuclear and mitochondrial DNA: The role of male-biased dispersal in a Holarctic sea duck

USGS Publications Warehouse

Peters, Jeffrey L.; Bolender, Kimberly A.; Pearce, John M.

2012-01-01

Genetic studies of waterfowl (Anatidae) have observed the full spectrum of mitochondrial (mt) DNA population divergence, from apparent panmixia to deep, reciprocally monophyletic lineages. Yet, these studies often found weak or no nuclear (nu) DNA structure, which was often attributed to male-biased gene flow, a common behaviour within this family. An alternative explanation for this ‘conflict’ is that the smaller effective population size and faster sorting rate of mtDNA relative to nuDNA lead to different signals of population structure. We tested these alternatives by sequencing 12 nuDNA introns for a Holarctic pair of waterfowl subspecies, the European goosander (Mergus merganser merganser) and the North American common merganser (M. m. americanus), which exhibit strong population structure in mtDNA. We inferred effective population sizes, gene flow and divergence times from published mtDNA sequences and simulated expected differentiation for nuDNA based on those histories. Between Europe and North America, nuDNA ФST was 3.4-fold lower than mtDNA ФST, a result consistent with differences in sorting rates. However, despite geographically structured and monophyletic mtDNA lineages within continents, nuDNA ФST values were generally zero and significantly lower than predicted. This between- and within-continent contrast held when comparing mtDNA and nuDNA among published studies of ducks. Thus, male-mediated gene flow is a better explanation than slower sorting rates for limited nuDNA differentiation within continents, which is also supported by nonmolecular data. This study illustrates the value of quantitatively testing discrepancies between mtDNA and nuDNA to reject the null hypothesis that conflict simply reflects different sorting rates.

The Serotonin Transporter Undergoes Constitutive Internalization and Is Primarily Sorted to Late Endosomes and Lysosomal Degradation*

PubMed Central

Rahbek-Clemmensen, Troels; Bay, Tina; Eriksen, Jacob; Gether, Ulrik; Jørgensen, Trine Nygaard

2014-01-01

The serotonin transporter (SERT) plays a critical role in regulating serotonin signaling by mediating reuptake of serotonin from the extracellular space. The molecular and cellular mechanisms controlling SERT levels in the membrane remain poorly understood. To study trafficking of the surface resident SERT, two functional epitope-tagged variants were generated. Fusion of a FLAG-tagged one-transmembrane segment protein Tac to the SERT N terminus generated a transporter with an extracellular epitope suited for trafficking studies (TacSERT). Likewise, a construct with an extracellular antibody epitope was generated by introducing an HA (hemagglutinin) tag in the extracellular loop 2 of SERT (HA-SERT). By using TacSERT and HA-SERT in antibody-based internalization assays, we show that SERT undergoes constitutive internalization in a dynamin-dependent manner. Confocal images of constitutively internalized SERT demonstrated that SERT primarily co-localized with the late endosomal/lysosomal marker Rab7, whereas little co-localization was observed with the Rab11, a marker of the “long loop” recycling pathway. This sorting pattern was distinct from that of a prototypical recycling membrane protein, the β2-adrenergic receptor. Furthermore, internalized SERT co-localized with the lysosomal marker LysoTracker and not with transferrin. The sorting pattern was further confirmed by visualizing internalization of SERT using the fluorescent cocaine analog JHC1-64 and by reversible and pulse-chase biotinylation assays showing evidence for lysosomal degradation of the internalized transporter. Finally, we found that SERT internalized in response to stimulation with 12-myristate 13-acetate co-localized primarily with Rab7- and LysoTracker-positive compartments. We conclude that SERT is constitutively internalized and that the internalized transporter is sorted mainly to degradation. PMID:24973209

Radiation Response of Cancer Stem-Like Cells From Established Human Cell Lines After Sorting for Surface Markers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Al-Assar, Osama; Muschel, Ruth J.; Mantoni, Tine S.

2009-11-15

Purpose: A subpopulation of cancer stem-like cells (CSLC) is hypothesized to exist in different cancer cell lines and to mediate radioresistance in solid tumors. Methods and Materials: Cells were stained for CSLC markers and sorted (fluorescence-activated cell sorter/magnetic beads) to compare foci and radiosensitivity of phosphorylated histone H2AX at Ser 139 (gamma-H2AX) in sorted vs. unsorted populations in eight cell lines from different organs. CSLC properties were examined using anchorage-independent growth and levels of activated Notch1. Validation consisted of testing tumorigenicity and postirradiation enrichment of CSLC in xenograft tumors. Results: The quantity of CSLC was generally in good agreement withmore » primary tumors. CSLC from MDA-MB-231 (breast) and Panc-1 and PSN-1 (both pancreatic) cells had fewer residual gamma-H2AX foci than unsorted cells, pointing to radioresistance of CSLC. However, only MDA-MB-231 CSLC were more radioresistant than unsorted cells. Furthermore, MDA-MB-231 CSLC showed enhanced anchorage-independent growth and overexpression of activated Notch1 protein. The expression of cancer stem cell surface markers in the MDA-MB-231 xenograft model was increased after exposure to fractionated radiation. In contrast to PSN-1 cells, a growth advantage for MDA-MB-231 CSLC xenograft tumors was found compared to tumors arising from unsorted cells. Conclusions: CSLC subpopulations showed no general radioresistant phenotype, despite the quantities of CSLC subpopulations shown to correspond relatively well in other reports. Likewise, CSLC characteristics were found in some but not all of the tested cell lines. The reported problems in testing for CSLC in cell lines may be overcome by additional techniques, beyond sorting for markers.« less

Prions amplify through degradation of the VPS10P sorting receptor sortilin.

PubMed

Uchiyama, Keiji; Tomita, Mitsuru; Yano, Masashi; Chida, Junji; Hara, Hideyuki; Das, Nandita Rani; Nykjaer, Anders; Sakaguchi, Suehiro

2017-06-01

Prion diseases are a group of fatal neurodegenerative disorders caused by prions, which consist mainly of the abnormally folded isoform of prion protein, PrPSc. A pivotal pathogenic event in prion disease is progressive accumulation of prions, or PrPSc, in brains through constitutive conformational conversion of the cellular prion protein, PrPC, into PrPSc. However, the cellular mechanism by which PrPSc is progressively accumulated in prion-infected neurons remains unknown. Here, we show that PrPSc is progressively accumulated in prion-infected cells through degradation of the VPS10P sorting receptor sortilin. We first show that sortilin interacts with PrPC and PrPSc and sorts them to lysosomes for degradation. Consistently, sortilin-knockdown increased PrPSc accumulation in prion-infected cells. In contrast, overexpression of sortilin reduced PrPSc accumulation in prion-infected cells. These results indicate that sortilin negatively regulates PrPSc accumulation in prion-infected cells. The negative role of sortilin in PrPSc accumulation was further confirmed in sortilin-knockout mice infected with prions. The infected mice had accelerated prion disease with early accumulation of PrPSc in their brains. Interestingly, sortilin was reduced in prion-infected cells and mouse brains. Treatment of prion-infected cells with lysosomal inhibitors, but not proteasomal inhibitors, increased the levels of sortilin. Moreover, sortilin was reduced following PrPSc becoming detectable in cells after infection with prions. These results indicate that PrPSc accumulation stimulates sortilin degradation in lysosomes. Taken together, these results show that PrPSc accumulation of itself could impair the sortilin-mediated sorting of PrPC and PrPSc to lysosomes for degradation by stimulating lysosomal degradation of sortilin, eventually leading to progressive accumulation of PrPSc in prion-infected cells.

Hemophagocytic Lymphohistiocytosis in a Young Child.

PubMed

Saikia, Uma Nahar; Gupta, Anju; Vignesh, Pandiarajan; Suri, Deepti; Singh, Mini P

2016-06-08

Hemophagocytic lymphohistiocytosis (HLH) is a multisystem disorder mediated by cytokine storm and is characterized by fever, pancytopenia and organomegaly coupled with laboratory features like hyperferritinemia, hypertriglyceridemia, hypofibrinogenemia and transaminitis. Etiology can be genetic or acquired such as infections, malignancy and autoimmune disorders. Diagnosis, identification of underlying etiology and management of HLH remain tough clinical puzzles to sort out for the managing physician. We report a clinico-pathological conference of a three-year-old boy who had such a presentation and succumbed during the hospital stay.

10 CFR 72.242 - Recordkeeping and reports.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 2 2010-01-01 2010-01-01 false Recordkeeping and reports. 72.242 Section 72.242 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSING REQUIREMENTS FOR THE INDEPENDENT STORAGE OF SPENT... Spent Fuel Storage Casks § 72.242 Recordkeeping and reports. (a) Each certificate holder or applicant...

The Role of the Neurofibromin-Syndecan-CASK Complex in the Regulation of Synaptic Ras-MAPK Signaling and Dendritic Spine Plasticity

DTIC Science & Technology

2006-02-01

Morgan, K., Hasz, D. E., Mao, Z., and Largaespada, D. A. (2005). Nf1 gene inactivation in acute myeloid leukemia cells confers cytarabine resistance through MAPK and mTOR pathways. Leukemia. VII. Appendices: None

Compromised fidelity of endocytic synaptic vesicle protein sorting in the absence of stonin 2

PubMed Central

Kononenko, Natalia L.; Diril, M. Kasim; Puchkov, Dmytro; Kintscher, Michael; Koo, Seong Joo; Pfuhl, Gerit; Winter, York; Wienisch, Martin; Klingauf, Jürgen; Breustedt, Jörg; Schmitz, Dietmar; Maritzen, Tanja; Haucke, Volker

2013-01-01

Neurotransmission depends on the exocytic fusion of synaptic vesicles (SVs) and their subsequent reformation either by clathrin-mediated endocytosis or budding from bulk endosomes. How synapses are able to rapidly recycle SVs to maintain SV pool size, yet preserve their compositional identity, is poorly understood. We demonstrate that deletion of the endocytic adaptor stonin 2 (Stn2) in mice compromises the fidelity of SV protein sorting, whereas the apparent speed of SV retrieval is increased. Loss of Stn2 leads to selective missorting of synaptotagmin 1 to the neuronal surface, an elevated SV pool size, and accelerated SV protein endocytosis. The latter phenotype is mimicked by overexpression of endocytosis-defective variants of synaptotagmin 1. Increased speed of SV protein retrieval in the absence of Stn2 correlates with an up-regulation of SV reformation from bulk endosomes. Our results are consistent with a model whereby Stn2 is required to preserve SV protein composition but is dispensable for maintaining the speed of SV recycling. PMID:23345427

Protein sorting by lipid phase-like domains supports emergent signaling function in B lymphocyte plasma membranes.

PubMed

Stone, Matthew B; Shelby, Sarah A; Núñez, Marcos F; Wisser, Kathleen; Veatch, Sarah L

2017-02-01

Diverse cellular signaling events, including B cell receptor (BCR) activation, are hypothesized to be facilitated by domains enriched in specific plasma membrane lipids and proteins that resemble liquid-ordered phase-separated domains in model membranes. This concept remains controversial and lacks direct experimental support in intact cells. Here, we visualize ordered and disordered domains in mouse B lymphoma cell membranes using super-resolution fluorescence localization microscopy, demonstrate that clustered BCR resides within ordered phase-like domains capable of sorting key regulators of BCR activation, and present a minimal, predictive model where clustering receptors leads to their collective activation by stabilizing an extended ordered domain. These results provide evidence for the role of membrane domains in BCR signaling and a plausible mechanism of BCR activation via receptor clustering that could be generalized to other signaling pathways. Overall, these studies demonstrate that lipid mediated forces can bias biochemical networks in ways that broadly impact signal transduction.

Principles and applications of flow cytometry and cell sorting in companion animal medicine.

PubMed

Wilkerson, Melinda J

2012-01-01

Flow cytometry measures multiple characteristic of single cells using light scatter properties and fluorescence properties of fluorescent probes with specificity to cellular constituents. The use of flow cytometry in the veterinary clinical laboratory has become more routine in veterinary diagnostic laboratories and institutions (http://www.vet.k-state.edu/depts/dmp/service/immunology/index.htm), and reference laboratories. The most common applications in small animal medicine includes quantitation of erythrocytes and leukocytes in automated hematology instruments, detection of antibodies to erythrocytes and platelets in cases of immune-mediated diseases, immunophenotyping of leukocytes and lymphocytes in immunodeficiency syndromes, or leukemias and lymphomas. DNA content analysis to identify aneuploidy or replicating cells in tumor preparations has not gained routine acceptance because of the variability of prognostic results. Other applications including cell sorting and multiplexing using microspheres are potential assays of the future once they become validated and the instrumentation footprint becomes more and more compact, less expensive, and easier to use.

COPI mediates recycling of an exocytic SNARE by recognition of a ubiquitin sorting signal

PubMed Central

Xu, Peng; Hankins, Hannah M; MacDonald, Chris; Erlinger, Samuel J; Frazier, Meredith N; Diab, Nicholas S; Piper, Robert C; Jackson, Lauren P; MacGurn, Jason A

2017-01-01

The COPI coat forms transport vesicles from the Golgi complex and plays a poorly defined role in endocytic trafficking. Here we show that COPI binds K63-linked polyubiquitin and this interaction is crucial for trafficking of a ubiquitinated yeast SNARE (Snc1). Snc1 is a v-SNARE that drives fusion of exocytic vesicles with the plasma membrane, and then recycles through the endocytic pathway to the Golgi for reuse in exocytosis. Removal of ubiquitin from Snc1, or deletion of a β'-COP subunit propeller domain that binds K63-linked polyubiquitin, disrupts Snc1 recycling causing aberrant accumulation in internal compartments. Moreover, replacement of the β'-COP propeller domain with unrelated ubiquitin-binding domains restores Snc1 recycling. These results indicate that ubiquitination, a modification well known to target membrane proteins to the lysosome or vacuole for degradation, can also function as recycling signal to sort a SNARE into COPI vesicles in a non-degradative pathway. PMID:29058666

CCC- and WASH-mediated endosomal sorting of LDLR is required for normal clearance of circulating LDL.

PubMed

Bartuzi, Paulina; Billadeau, Daniel D; Favier, Robert; Rong, Shunxing; Dekker, Daphne; Fedoseienko, Alina; Fieten, Hille; Wijers, Melinde; Levels, Johannes H; Huijkman, Nicolette; Kloosterhuis, Niels; van der Molen, Henk; Brufau, Gemma; Groen, Albert K; Elliott, Alison M; Kuivenhoven, Jan Albert; Plecko, Barbara; Grangl, Gernot; McGaughran, Julie; Horton, Jay D; Burstein, Ezra; Hofker, Marten H; van de Sluis, Bart

2016-03-11

The low-density lipoprotein receptor (LDLR) plays a pivotal role in clearing atherogenic circulating low-density lipoprotein (LDL) cholesterol. Here we show that the COMMD/CCDC22/CCDC93 (CCC) and the Wiskott-Aldrich syndrome protein and SCAR homologue (WASH) complexes are both crucial for endosomal sorting of LDLR and for its function. We find that patients with X-linked intellectual disability caused by mutations in CCDC22 are hypercholesterolaemic, and that COMMD1-deficient dogs and liver-specific Commd1 knockout mice have elevated plasma LDL cholesterol levels. Furthermore, Commd1 depletion results in mislocalization of LDLR, accompanied by decreased LDL uptake. Increased total plasma cholesterol levels are also seen in hepatic COMMD9-deficient mice. Inactivation of the CCC-associated WASH complex causes LDLR mislocalization, increased lysosomal degradation of LDLR and impaired LDL uptake. Furthermore, a mutation in the WASH component KIAA0196 (strumpellin) is associated with hypercholesterolaemia in humans. Altogether, this study provides valuable insights into the mechanisms regulating cholesterol homeostasis and LDLR trafficking.

Clathrin Terminal Domain-Ligand Interactions Regulate Sorting of Mannose 6-Phosphate Receptors Mediated by AP-1 and GGA Adaptors*

PubMed Central

Stahlschmidt, Wiebke; Robertson, Mark J.; Robinson, Phillip J.; McCluskey, Adam; Haucke, Volker

2014-01-01

Clathrin plays important roles in intracellular membrane traffic including endocytosis of plasma membrane proteins and receptors and protein sorting between the trans-Golgi network (TGN) and endosomes. Whether clathrin serves additional roles in receptor recycling, degradative sorting, or constitutive secretion has remained somewhat controversial. Here we have used acute pharmacological perturbation of clathrin terminal domain (TD) function to dissect the role of clathrin in intracellular membrane traffic. We report that internalization of major histocompatibility complex I (MHCI) is inhibited in cells depleted of clathrin or its major clathrin adaptor complex 2 (AP-2), a phenotype mimicked by application of Pitstop® inhibitors of clathrin TD function. Hence, MHCI endocytosis occurs via a clathrin/AP-2-dependent pathway. Acute perturbation of clathrin also impairs the dynamics of intracellular clathrin/adaptor complex 1 (AP-1)- or GGA (Golgi-localized, γ-ear-containing, Arf-binding protein)-coated structures at the TGN/endosomal interface, resulting in the peripheral dispersion of mannose 6-phosphate receptors. By contrast, secretory traffic of vesicular stomatitis virus G protein, recycling of internalized transferrin from endosomes, or degradation of EGF receptor proceeds unperturbed in cells with impaired clathrin TD function. These data indicate that clathrin is required for the function of AP-1- and GGA-coated carriers at the TGN but may be dispensable for outward traffic en route to the plasma membrane. PMID:24407285

Efficient genome editing by FACS enrichment of paired D10A Cas9 nickases coupled with fluorescent proteins.

PubMed

Gopalappa, Ramu; Song, Myungjae; Chandrasekaran, Arun Pandian; Das, Soumyadip; Haq, Saba; Koh, Hyun Chul; Ramakrishna, Suresh

2018-05-31

Targeted genome editing by clustered regularly interspaced short palindromic repeats (CRISPR-Cas9) raised concerns over off-target effects. The use of double-nicking strategy using paired Cas9 nickase has been developed to minimize off-target effects. However, it was reported that the efficiency of paired nickases were comparable or lower than that of either corresponding nuclease alone. Recently, we conducted a systematic comparison of the efficiencies of several paired Cas9 with their corresponding Cas9 nucleases and showed that paired D10A Cas9 nickases are sometimes more efficient than individual nucleases for gene disruption. However, sometimes the designed paired Cas9 nickases exhibited significantly lower mutation frequencies than nucleases, hampering the generation of cells containing paired Cas9 nickase-induced mutations. Here we implemented IRES peptide-conjugation of fluorescent protein to Cas9 nickase and subjected for fluorescence-activated cell sorting. The sorted cell populations are highly enriched with cells containing paired Cas9 nickase-induced mutations, by a factor of up to 40-fold as compared with the unsorted population. Furthermore, gene-disrupted single cell clones using paired nickases followed by FACS sorting strategy were generated highly efficiently, without compromising with its low off-target effects. We envision that our fluorescent protein coupled paired nickase-mediated gene disruption, facilitating efficient and highly specific genome editing in medical research.

Programmable In Vitro Coencapsidation of Guest Proteins for Intracellular Delivery by Virus-like Particles.

PubMed

Dashti, Noor H; Abidin, Rufika S; Sainsbury, Frank

2018-05-22

Bioinspired self-sorting and self-assembling systems using engineered versions of natural protein cages are being developed for biocatalysis and therapeutic delivery. The packaging and intracellular delivery of guest proteins is of particular interest for both in vitro and in vivo cell engineering. However, there is a lack of bionanotechnology platforms that combine programmable guest protein encapsidation with efficient intracellular uptake. We report a minimal peptide anchor for in vivo self-sorting of cargo-linked capsomeres of murine polyomavirus (MPyV) that enables controlled encapsidation of guest proteins by in vitro self-assembly. Using Förster resonance energy transfer, we demonstrate the flexibility in this system to support coencapsidation of multiple proteins. Complementing these ensemble measurements with single-particle analysis by super-resolution microscopy shows that the stochastic nature of coencapsidation is an overriding principle. This has implications for the design and deployment of both native and engineered self-sorting encapsulation systems and for the assembly of infectious virions. Taking advantage of the encoded affinity for sialic acids ubiquitously displayed on the surface of mammalian cells, we demonstrate the ability of self-assembled MPyV virus-like particles to mediate efficient delivery of guest proteins to the cytosol of primary human cells. This platform for programmable coencapsidation and efficient cytosolic delivery of complementary biomolecules therefore has enormous potential in cell engineering.

A role for microtubules in sorting endocytic vesicles in rat hepatocytes.

PubMed Central

Goltz, J S; Wolkoff, A W; Novikoff, P M; Stockert, R J; Satir, P

1992-01-01

The vectorial nature of hepatocyte receptor-mediated endocytosis (RME) and its susceptibility to cytoskeletal disruptors has suggested that a polarized network of microtubules plays a vital role in directed movement during sorting. Using as markers a well-known ligand, asialoorosomucoid, and its receptor, we have isolated endocytic vesicles that bind directly to and interact with stabilized endogenous hepatocyte microtubules at specific times during a synchronous, experimentally initiated, single wave of RME. Both ligand- and receptor-containing vesicles copelleted with microtubules in the absence of ATP but did not pellet under similar conditions when microtubules were not polymerized. When 5 mM ATP was added to preparations of microtubule-bound vesicles, ligand-containing vesicles were released into the supernatant, while receptor-containing vesicles remained immobilized on the microtubules. Release of ligand-containing vesicles from microtubules was prevented by monensin treatment during the endocytic wave. Several proteins, including the microtubule motor protein cytoplasmic dynein, were present in these preparations and were released from microtubule pellets by ATP addition concomitantly with ligand. These results suggest that receptor domains within the endosome can be immobilized by attachment to microtubules so that, following monensin-sensitive dissociation of ligand from receptor, ligand-containing vesicles can be pulled along microtubules away from the receptor domains by a motor molecule, such as cytoplasmic dynein, thereby delineating sorting. Images PMID:1353884

27 CFR 25.35 - Tanks.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Tanks. 25.35 Section 25.35... TREASURY LIQUORS BEER Construction and Equipment Equipment § 25.35 Tanks. Each stationary tank, vat, cask... contents of tanks or containers in lieu of providing each tank or container with a measuring device. (Sec...

27 CFR 25.35 - Tanks.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 27 Alcohol, Tobacco Products and Firearms 1 2013-04-01 2013-04-01 false Tanks. 25.35 Section 25.35... TREASURY ALCOHOL BEER Construction and Equipment Equipment § 25.35 Tanks. Each stationary tank, vat, cask... contents of tanks or containers in lieu of providing each tank or container with a measuring device. (Sec...

10 CFR 72.234 - Conditions of approval.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 2 2010-01-01 2010-01-01 false Conditions of approval. 72.234 Section 72.234 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSING REQUIREMENTS FOR THE INDEPENDENT STORAGE OF SPENT... Spent Fuel Storage Casks § 72.234 Conditions of approval. (a) The certificate holder and applicant for a...

10 CFR 72.232 - Inspection and tests.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 2 2010-01-01 2010-01-01 false Inspection and tests. 72.232 Section 72.232 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSING REQUIREMENTS FOR THE INDEPENDENT STORAGE OF SPENT NUCLEAR FUEL... Storage Casks § 72.232 Inspection and tests. (a) The certificate holder and applicant for a CoC shall...

10 CFR 72.248 - Safety analysis report updating.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 2 2010-01-01 2010-01-01 false Safety analysis report updating. 72.248 Section 72.248 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSING REQUIREMENTS FOR THE INDEPENDENT STORAGE OF... Approval of Spent Fuel Storage Casks § 72.248 Safety analysis report updating. (a) Each certificate holder...

75 FR 33736 - List of Approved Spent Fuel Storage Casks: MAGNASTOR System, Revision 1

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-15

... Management Programs, U.S. Nuclear Regulatory Commission, Washington, DC 20555-0001, telephone (301) 415- 6219..., 11555 Rockville Pike, Rockville, Maryland. NRC's Agencywide Documents Access and Management System... M. McCausland, Office of Federal and State Materials and Environmental Management Programs, U.S...

76 FR 9381 - Notice of Availability of Interim Staff Guidance Documents for Spent Fuel Storage Casks

Federal Register 2010, 2011, 2012, 2013, 2014

2011-02-17

.... FOR FURTHER INFORMATION CONTACT: Matthew Gordon, Structural Mechanics and Materials Branch, Division... a fee. Comments and questions on ISG-23 should be directed to Matthew Gordon, Structural Mechanics..., 2011. For the U.S. Nuclear Regulatory Commission. Michele Sampson, Acting Chief, Structural Mechanics...

27 CFR 25.35 - Tanks.

Code of Federal Regulations, 2012 CFR

2012-04-01

... TREASURY LIQUORS BEER Construction and Equipment Equipment § 25.35 Tanks. Each stationary tank, vat, cask or other container used, or intended for use, as a receptacle for wort, beer or concentrate produced from beer shall: (a) Be durably marked with a serial number and capacity; and (b) Be equipped with a...

27 CFR 25.35 - Tanks.

Code of Federal Regulations, 2014 CFR

2014-04-01

... TREASURY ALCOHOL BEER Construction and Equipment Equipment § 25.35 Tanks. Each stationary tank, vat, cask or other container used, or intended for use, as a receptacle for wort, beer or concentrate produced from beer shall: (a) Be durably marked with a serial number and capacity; and (b) Be equipped with a...

75 FR 41404 - List of Approved Spent Fuel Storage Casks: NUHOMS®

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-16

.... The NRC is taking this action because the applicant identified that a certain Technical Specification (TS) for Boral characterization was not written precisely. Specifically, the requirements for meeting... changes to the technical specifications. The NRC also published a direct final rule on May 6, 2010 (75 FR...

75 FR 41369 - List of Approved Spent Fuel Storage Casks: NUHOMS® HD Revision 1; Withdrawal

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-16

...) Number 1030. The NRC is taking this action because the applicant identified that a certain Technical Specification (TS) for Boral characterization was not written precisely and in a manner that could be readily... cavity water removal operations, and making [[Page 41370

76 FR 2243 - List of Approved Spent Fuel Storage Casks: NUHOMS ® HD System Revision 1

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-13

... the requirements of reconstituted fuel assemblies; add requirements to qualify metal matrix composite... requirements to qualify metal matrix composite neutron absorbers with integral aluminum cladding; clarify the... requirements to qualify metal matrix composite neutron absorbers with integral aluminum cladding; clarify the...

78 FR 3853 - Retrievability, Cladding Integrity and Safe Handling of Spent Fuel at an Independent Spent Fuel...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-01-17

... requirement that loaded storage casks also meet transportation requirements. Integration of storage and... transported from the storage location. As part of its evaluation of integration and compatibility between... evaluating compatibility of storage and transportation regulations. As part of its evaluation of integration...

27 CFR 25.35 - Tanks.

Code of Federal Regulations, 2011 CFR

2011-04-01

... TREASURY LIQUORS BEER Construction and Equipment Equipment § 25.35 Tanks. Each stationary tank, vat, cask or other container used, or intended for use, as a receptacle for wort, beer or concentrate produced from beer shall: (a) Be durably marked with a serial number and capacity; and (b) Be equipped with a...

OCRWM Bulletin: Westinghouse begins designing multi-purpose canister

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1995-09-01

This publication consists of two parts: OCRWM (Office of Civilian Radioactive Waste Management) Bulletin; and Of Mountains & Science which has articles on the Yucca Mountain project. The OCRWM provides information about OCRWM activities and in this issue has articles on multi-purpose canister design, and transportation cask trailer.

78 FR 79021 - Sunshine Act Meeting Notice

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-27

... NUCLEAR REGULATORY COMMISSION [NRC-2013-0001] Sunshine Act Meeting Notice DATES: Weeks of December... meetings scheduled for the week of December 30, 2013. Week of January 6, 2014--Tentative Monday, January 6... to Dry Casks (Public Meeting) (Contact: Kevin Witt, 301-415-2145). This meeting will be webcast live...

27 CFR 26.40 - Marking containers of distilled spirits.

Code of Federal Regulations, 2010 CFR

2010-04-01

... spirits. The distiller, rectifier, or bottler shall serially number each case, barrel, cask, or similar... the container, the distiller, rectifier, or bottler shall plainly print, stamp, or stencil with..., rectifier, or bottler. (b) The brand name and kind of liquor; (c) The wine and proof gallon contents; or...

Cytokines and innate inflammation in the pathogenesis of human traumatic brain injury.

PubMed

Helmy, Adel; De Simoni, Maria-Grazia; Guilfoyle, Mathew R; Carpenter, Keri L H; Hutchinson, Peter J

2011-11-01

There is an increasing recognition that following traumatic brain injury, a cascade of inflammatory mediators is produced, and contributes to the pathological consequences of central nervous system injury. This review summarises the key literature from pre-clinical models that underlies our understanding of innate inflammation following traumatic brain injury before focussing on the growing evidence from human studies. In addition, the underlying molecular mediators responsible for blood brain barrier dysfunction have been discussed. In particular, we have highlighted the different sampling methodologies available and the difficulties in interpreting human data of this sort. Ultimately, understanding the innate inflammatory response to traumatic brain injury may provide a therapeutic avenue in the treatment of central nervous system disease. Copyright © 2011 Elsevier Ltd. All rights reserved.

Transposon-mediated generation of BCR-ABL1-expressing transgenic cell lines for unbiased sensitivity testing of tyrosine kinase inhibitors

PubMed Central

Berkowitsch, Bettina; Koenig, Margit; Haas, Oskar A.; Hoermann, Gregor; Valent, Peter; Lion, Thomas

2016-01-01

Point mutations in the ABL1 kinase domain are an important mechanism of resistance to tyrosine kinase inhibitors (TKI) in BCR-ABL1-positive and, as recently shown, BCR-ABL1-like leukemias. The cell line Ba/F3 lentivirally transduced with mutant BCR-ABL1 constructs is widely used for in vitro sensitivity testing and response prediction to tyrosine kinase inhibitors. The transposon-based Sleeping Beauty system presented offers several advantages over lentiviral transduction including the absence of biosafety issues, faster generation of transgenic cell lines, and greater efficacy in introducing large gene constructs. Nevertheless, both methods can mediate multiple insertions in the genome. Here we show that multiple BCR-ABL1 insertions result in elevated IC50 levels for individual TKIs, thus overestimating the actual resistance of mutant subclones. We have therefore established flow-sorting-based fractionation of BCR-ABL1-transformed Ba/F3 cells facilitating efficient enrichment of cells carrying single-site insertions, as demonstrated by FISH-analysis. Fractions of unselected Ba/F3 cells not only showed a greater number of BCR-ABL1 hybridization signals, but also revealed higher IC50 values for the TKIs tested. The data presented highlight the need to carefully select transfected cells by flow-sorting, and to control the insertion numbers by FISH and real-time PCR to permit unbiased in vitro testing of drug resistance. PMID:27801667

Lentiviral gene ontology (LeGO) vectors equipped with novel drug-selectable fluorescent proteins: new building blocks for cell marking and multi-gene analysis.

PubMed

Weber, K; Mock, U; Petrowitz, B; Bartsch, U; Fehse, B

2010-04-01

Vector-encoded fluorescent proteins (FPs) facilitate unambiguous identification or sorting of gene-modified cells by fluorescence-activated cell sorting (FACS). Exploiting this feature, we have recently developed lentiviral gene ontology (LeGO) vectors (www.LentiGO-Vectors.de) for multi-gene analysis in different target cells. In this study, we extend the LeGO principle by introducing 10 different drug-selectable FPs created by fusing one of the five selection marker (protecting against blasticidin, hygromycin, neomycin, puromycin and zeocin) and one of the five FP genes (Cerulean, eGFP, Venus, dTomato and mCherry). All tested fusion proteins allowed both fluorescence-mediated detection and drug-mediated selection of LeGO-transduced cells. Newly generated codon-optimized hygromycin- and neomycin-resistance genes showed improved expression as compared with their ancestors. New LeGO constructs were produced at titers >10(6) per ml (for non-concentrated supernatants). We show efficient combinatorial marking and selection of various cells, including mesenchymal stem cells, simultaneously transduced with different LeGO constructs. Inclusion of the cytomegalovirus early enhancer/chicken beta-actin promoter into LeGO vectors facilitated robust transgene expression in and selection of neural stem cells and their differentiated progeny. We suppose that the new drug-selectable markers combining advantages of FACS and drug selection are well suited for numerous applications and vector systems. Their inclusion into LeGO vectors opens new possibilities for (stem) cell tracking and functional multi-gene analysis.

Evolutionary Changes on the Way to Clathrin-Mediated Endocytosis in Animals

PubMed Central

Dergai, Mykola; Iershov, Anton; Novokhatska, Olga; Pankivskyi, Serhii; Rynditch, Alla

2016-01-01

Endocytic pathways constitute an evolutionarily ancient system that significantly contributed to the eukaryotic cell architecture and to the diversity of cell type–specific functions and signaling cascades, in particular of metazoans. Here we used comparative proteomic studies to analyze the universal internalization route in eukaryotes, clathrin-mediated endocytosis (CME), to address the issues of how this system evolved and what are its specific features. Among 35 proteins crucially required for animal CME, we identified a subset of 22 proteins common to major eukaryotic branches and 13 gradually acquired during evolution. Based on exploration of structure–function relationship between conserved homologs in sister, distantly related and early diverged branches, we identified novel features acquired during evolution of endocytic proteins on the way to animals: Elaborated way of cargo recruitment by multiple sorting proteins, structural changes in the core endocytic complex AP2, the emergence of the Fer/Cip4 homology domain-only protein/epidermal growth factor receptor substrate 15/intersectin functional complex as an additional interaction hub and activator of AP2, as well as changes in late endocytic stages due to recruitment of dynamin/sorting nexin 9 complex and involvement of the actin polymerization machinery. The evolutionary reconstruction showed the basis of the CME process and its subsequent step-by-step development. Documented changes imply more precise regulation of the pathway, as well as CME specialization for the uptake of specific cargoes and cell type-specific functions. PMID:26872775

Multi-risk infants: predicting attachment security from sociodemographic, psychosocial, and health risk among African-American preterm infants.

PubMed

Candelaria, Margo; Teti, Douglas M; Black, Maureen M

2011-08-01

Ecological and transactional theories link child outcomes to accumulated risk. This study hypothesized that cumulative risk was negatively related to attachment, and that maternal sensitivity mediated linkages between risk and attachment. One hundred and twelve high-risk African-American premature infant-mother dyads participated. Psychosocial (maternal depression, stress and self-efficacy) and sociodemographic risk (poverty, maternal education, marital status) were maternal self-report (0-4 months). Infant health risk was obtained from hospital charts. Infant-mother attachment (12 months) and maternal sensitivity (4 months) were assessed with Q-sort measures. Psychosocial and sociodemographic risk, but not infant health risk, negatively related to attachment. Both were mediated by maternal sensitivity. The impact of risk domains on attachment security was mediated by maternal sensitivity. Results emphasize the need for early intervention programs targeting premature infants to identify and address environmental and personal factors that place parenting at risk. © 2011 The Authors. Journal of Child Psychology and Psychiatry © 2011 Association for Child and Adolescent Mental Health.

Alterations in CDH15 and KIRREL3 in Patients with Mild to Severe Intellectual Disability

PubMed Central

Bhalla, Kavita; Luo, Yue; Buchan, Tim; Beachem, Michael A.; Guzauskas, Gregory F.; Ladd, Sydney; Bratcher, Shelly J.; Schroer, Richard J.; Balsamo, Janne; DuPont, Barbara R.; Lilien, Jack; Srivastava, Anand K.

2008-01-01

Cell-adhesion molecules play critical roles in brain development, as well as maintaining synaptic structure, function, and plasticity. Here we have found the disruption of two genes encoding putative cell-adhesion molecules, CDH15 (cadherin superfamily) and KIRREL3 (immunoglobulin superfamily), by a chromosomal translocation t(11;16) in a female patient with intellectual disability (ID). We screened coding regions of these two genes in a cohort of patients with ID and controls and identified four nonsynonymous CDH15 variants and three nonsynonymous KIRREL3 variants that appear rare and unique to ID. These variations altered highly conserved residues and were absent in more than 600 unrelated patients with ID and 800 control individuals. Furthermore, in vivo expression studies showed that three of the CDH15 variations adversely altered its ability to mediate cell-cell adhesion. We also show that in neuronal cells, human KIRREL3 colocalizes and interacts with the synaptic scaffolding protein, CASK, recently implicated in X-linked brain malformation and ID. Taken together, our data suggest that alterations in CDH15 and KIRREL3, either alone or in combination with other factors, could play a role in phenotypic expression of ID in some patients. PMID:19012874

10 CFR 72.248 - Safety analysis report updating.

Code of Federal Regulations, 2014 CFR

2014-01-01

... appropriate, the last update to the FSAR under this section. The update shall include the effects 1 of: 1... for a spent fuel storage cask design shall update periodically, as provided in paragraph (b) of this... the issued Certificate of Compliance (CoC). (b) Each update shall contain all the changes necessary to...

10 CFR 72.248 - Safety analysis report updating.

Code of Federal Regulations, 2013 CFR

2013-01-01

... appropriate, the last update to the FSAR under this section. The update shall include the effects 1 of: 1... for a spent fuel storage cask design shall update periodically, as provided in paragraph (b) of this... the issued Certificate of Compliance (CoC). (b) Each update shall contain all the changes necessary to...

10 CFR 72.248 - Safety analysis report updating.

Code of Federal Regulations, 2012 CFR

2012-01-01

... appropriate, the last update to the FSAR under this section. The update shall include the effects 1 of: 1... for a spent fuel storage cask design shall update periodically, as provided in paragraph (b) of this... the issued Certificate of Compliance (CoC). (b) Each update shall contain all the changes necessary to...

10 CFR 72.248 - Safety analysis report updating.

Code of Federal Regulations, 2011 CFR

2011-01-01

... appropriate, the last update to the FSAR under this section. The update shall include the effects 1 of: 1... for a spent fuel storage cask design shall update periodically, as provided in paragraph (b) of this... the issued Certificate of Compliance (CoC). (b) Each update shall contain all the changes necessary to...

10 CFR 72.48 - Changes, tests, and experiments.

Code of Federal Regulations, 2011 CFR

2011-01-01

... facility or spent fuel storage cask design, of changes in procedures, and of tests and experiments made... 10 Energy 2 2011-01-01 2011-01-01 false Changes, tests, and experiments. 72.48 Section 72.48... Issuance and Conditions of License § 72.48 Changes, tests, and experiments. (a) Definitions for the...

10 CFR 72.48 - Changes, tests, and experiments.

Code of Federal Regulations, 2010 CFR

2010-01-01

... facility or spent fuel storage cask design, of changes in procedures, and of tests and experiments made... 10 Energy 2 2010-01-01 2010-01-01 false Changes, tests, and experiments. 72.48 Section 72.48... Issuance and Conditions of License § 72.48 Changes, tests, and experiments. (a) Definitions for the...

76 FR 70374 - List of Approved Spent Fuel Storage Casks: MAGNASTOR® System, Revision 2

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-14

... Trussell, Office of Federal and State Materials and Environmental Management Programs, U.S. Nuclear... Management System (ADAMS): Publicly available documents created or received at the NRC are available online... protection of public health and safety continues to be ensured. The direct final rule will become effective...

27 CFR 26.207 - Destruction of marks and brands.

Code of Federal Regulations, 2010 CFR

2010-04-01

... brands. 26.207 Section 26.207 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE... Products Coming Into the United States From the Virgin Islands § 26.207 Destruction of marks and brands. The marks, brands, and serial numbers required by this part to be placed on barrels, casks, or similar...

27 CFR 26.41 - Destruction of marks and brands.

Code of Federal Regulations, 2010 CFR

2010-04-01

... brands. 26.41 Section 26.41 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE... Products Coming Into the United States From Puerto Rico § 26.41 Destruction of marks and brands. The marks, brands, and serial numbers required by this part to be placed on barrels, casks, or similar containers...

Preliminary risk assessment for nuclear waste disposal in space, volume 2

NASA Technical Reports Server (NTRS)

Rice, E. E.; Denning, R. S.; Friedlander, A. L.

1982-01-01

Safety guidelines are presented. Waste form, waste processing and payload fabrication facilities, shipping casks and ground transport vehicles, payload primary container/core, radiation shield, reentry systems, launch site facilities, uprooted space shuttle launch vehicle, Earth packing orbits, orbit transfer systems, and space destination are discussed. Disposed concepts and risks are then discussed.

9. DETAIL VIEW OF BRIDGE CRANE ON WEST SIDE OF ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

9. DETAIL VIEW OF BRIDGE CRANE ON WEST SIDE OF BUILDING. CAMERA FACING NORTHEAST. CONTAMINATED AIR FILTERS LOADED IN TRANSPORT CASKS WERE TRANSFERRED TO VEHICLES AND SENT TO RADIOACTIVE WASTE MANAGEMENT COMPLEX FOR STORAGE. INEEL PROOF NUMBER HD-17-1. - Idaho National Engineering Laboratory, Old Waste Calcining Facility, Scoville, Butte County, ID

78 FR 74188 - Sunshine Act Meetings Notice

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-10

... Casks (Public Meeting) (Contact: Kevin Witt, 301-415-2145) This meeting will be Web cast live at the Web... Weather Events (Public Meeting) (Contact: George Wilson, 301-415-1711) This meeting will be Web cast live... will be Web cast live at the Web address-- http://www.nrc.gov/ . Week of January 13, 2014--Tentative...

Between wilderness and the middle landscape: A rocky road

Treesearch

Lisi Krall

2007-01-01

Wilderness preservation, as one branch of conservation, demonstrates a decidedly different cultural ethos than the utilitarian branch. Thus, preservation and utilitarian conservation represent different habits of thought fermenting in the cask of l9th century economic evolution. More specifically, the utilitarian branch of conservation can easily be viewed as an...

10 CFR 72.103 - Geological and seismological characteristics for applications for dry cask modes of storage on or...

Code of Federal Regulations, 2010 CFR

2010-01-01

..., and that are not in areas of known seismic activity, a standardized design earthquake ground motion... motion, tectonic surface deformation, nontectonic deformation, earthquake recurrence rates, fault... of the Design Earthquake Ground Motion (DE). The DE for the site is characterized by both horizontal...

10 CFR 72.103 - Geological and seismological characteristics for applications for dry cask modes of storage on or...

Code of Federal Regulations, 2011 CFR

2011-01-01

..., and that are not in areas of known seismic activity, a standardized design earthquake ground motion... motion, tectonic surface deformation, nontectonic deformation, earthquake recurrence rates, fault... of the Design Earthquake Ground Motion (DE). The DE for the site is characterized by both horizontal...

10 CFR 72.103 - Geological and seismological characteristics for applications for dry cask modes of storage on or...

Code of Federal Regulations, 2013 CFR

2013-01-01

..., and that are not in areas of known seismic activity, a standardized design earthquake ground motion... motion, tectonic surface deformation, nontectonic deformation, earthquake recurrence rates, fault... of the Design Earthquake Ground Motion (DE). The DE for the site is characterized by both horizontal...

10 CFR 72.103 - Geological and seismological characteristics for applications for dry cask modes of storage on or...

Code of Federal Regulations, 2014 CFR

2014-01-01

..., and that are not in areas of known seismic activity, a standardized design earthquake ground motion... motion, tectonic surface deformation, nontectonic deformation, earthquake recurrence rates, fault... of the Design Earthquake Ground Motion (DE). The DE for the site is characterized by both horizontal...

10 CFR 72.103 - Geological and seismological characteristics for applications for dry cask modes of storage on or...

Code of Federal Regulations, 2012 CFR

2012-01-01

..., and that are not in areas of known seismic activity, a standardized design earthquake ground motion... motion, tectonic surface deformation, nontectonic deformation, earthquake recurrence rates, fault... of the Design Earthquake Ground Motion (DE). The DE for the site is characterized by both horizontal...

DOE Office of Scientific and Technical Information (OSTI.GOV)

Finch, Robert J.; Smartt, Heidi A.; Haddal, Risa

Once a geological repository has begun operations, the encapsulation and disposal of spent fuel will be performed as a continuous, industrial-scale series of processes, during which time safeguards seals will be applied to transportation casks before shipment from an encapsulation plant, and then verified and removed following receipt at the repository. These operations will occur approximately daily during several decades of Sweden's repository operation; however, requiring safeguards inspectors to perform the application, verification, and removal of every seal would be an onerous burden on International Atomic Energy Agency's (IAEA's) resources. Current IAEA practice includes allowing operators to either apply sealsmore » or remove them, but not both, so the daily task of either applying or verifying and removing would still require continuous presence of IAEA inspectors at one site at least. Of special importance is the inability to re-verify cask or canisters from which seals have been removed and the canisters emplaced underground. Successfully designing seals that can be applied, verified and removed by an operator with IAEA approval could impact more than repository shipments, but other applications as well, potentially reducing inspector burdens for a wide range of such duties.« less

Baseline Design Compliance Matrix for the Rotary Mode Core Sampling System

DOE Office of Scientific and Technical Information (OSTI.GOV)

LECHELT, J.A.

2000-10-17

The purpose of the design compliance matrix (DCM) is to provide a single-source document of all design requirements associated with the fifteen subsystems that make up the rotary mode core sampling (RMCS) system. It is intended to be the baseline requirement document for the RMCS system and to be used in governing all future design and design verification activities associated with it. This document is the DCM for the RMCS system used on Hanford single-shell radioactive waste storage tanks. This includes the Exhauster System, Rotary Mode Core Sample Trucks, Universal Sampling System, Diesel Generator System, Distribution Trailer, X-Ray Cart System,more » Breathing Air Compressor, Nitrogen Supply Trailer, Casks and Cask Truck, Service Trailer, Core Sampling Riser Equipment, Core Sampling Support Trucks, Foot Clamp, Ramps and Platforms and Purged Camera System. Excluded items are tools such as light plants and light stands. Other items such as the breather inlet filter are covered by a different design baseline. In this case, the inlet breather filter is covered by the Tank Farms Design Compliance Matrix.« less

Differential effects of BDNF and neurotrophin 4 (NT4) on endocytic sorting of TrkB receptors.

PubMed

Proenca, Catia C; Song, Minseok; Lee, Francis S

2016-08-01

Neurotrophins are a family of growth factors playing key roles in the survival, development, and function of neurons. The neurotrophins brain-derived neurotrophic factor (BDNF) and NT4 both bind to and activate TrkB receptors, however, they mediate distinct neuronal functions. The molecular mechanism of how TrkB activation by BDNF and NT4 leads to diverse outcomes is unknown. Here, we report that BDNF and NT4 lead to differential endocytic sorting of TrkB receptors resulting in diverse biological functions in cultured cortical neurons. Fluorescent microscopy and surface biotinylation experiments showed that both neurotrophins stimulate internalization of TrkB with similar kinetics. Exposure to BDNF for 2-3 h reduced the surface pool of TrkB receptors to half, whereas a longer treatment (4-5 h) with NT4 was necessary to achieve a similar level of down-regulation. Although BDNF and NT4 induced TrkB phosphorylation with similar intensities, BDNF induced more rapid ubiquitination and degradation of TrkB than NT4. Interestingly, TrkB receptor ubiquitination by these ligands have substantially different pH sensitivities, resulting in varying degrees of receptor ubiquitination at lower pH levels. Consequently, NT4 was capable of maintaining longer sustained downstream signaling activation that correlated with reduced TrkB ubiquitination at endosomal pH. Thus, by leading to altered endocytic trafficking itineraries for TrkB receptors, BDNF and NT4 elicit differential TrkB signaling in terms of duration, intensity, and specificity, which may contribute to their functional differences in vivo. The neurotrophins, brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT4), both bind to and activate TrkB receptors, however, they mediate distinct neuronal functions. Here, we propose that BDNF and NT4 lead to differential endocytic sorting of TrkB receptors resulting in diverse biological functions. BDNF induces more rapid ubiquitination and degradation of TrkB than NT4. Consequently, NT4 is capable of maintaining more sustained signaling downstream of TrkB receptors. © 2016 International Society for Neurochemistry.

VGLUT2 Trafficking Is Differentially Regulated by Adaptor Proteins AP-1 and AP-3

PubMed Central

Li, Haiyan; Santos, Magda S.; Park, Chihyung K.; Dobry, Yuriy; Voglmaier, Susan M.

2017-01-01

Release of the major excitatory neurotransmitter glutamate by synaptic vesicle exocytosis depends on glutamate loading into synaptic vesicles by vesicular glutamate transporters (VGLUTs). The two principal isoforms, VGLUT1 and 2, exhibit a complementary pattern of expression in adult brain that broadly distinguishes cortical (VGLUT1) and subcortical (VGLUT2) systems, and correlates with distinct physiological properties in synapses expressing these isoforms. Differential trafficking of VGLUT1 and 2 has been suggested to underlie their functional diversity. Increasing evidence suggests individual synaptic vesicle proteins use specific sorting signals to engage specialized biochemical mechanisms to regulate their recycling. We observed that VGLUT2 recycles differently in response to high frequency stimulation than VGLUT1. Here we further explore the trafficking of VGLUT2 using a pHluorin-based reporter, VGLUT2-pH. VGLUT2-pH exhibits slower rates of both exocytosis and endocytosis than VGLUT1-pH. VGLUT2-pH recycling is slower than VGLUT1-pH in both hippocampal neurons, which endogenously express mostly VGLUT1, and thalamic neurons, which endogenously express mostly VGLUT2, indicating that protein identity, not synaptic vesicle membrane or neuronal cell type, controls sorting. We characterize sorting signals in the C-terminal dileucine-like motif, which plays a crucial role in VGLUT2 trafficking. Disruption of this motif abolishes synaptic targeting of VGLUT2 and essentially eliminates endocytosis of the transporter. Mutational and biochemical analysis demonstrates that clathrin adaptor proteins (APs) interact with VGLUT2 at the dileucine-like motif. VGLUT2 interacts with AP-2, a well-studied adaptor protein for clathrin mediated endocytosis. In addition, VGLUT2 also interacts with the alternate adaptors, AP-1 and AP-3. VGLUT2 relies on distinct recycling mechanisms from VGLUT1. Abrogation of these differences by pharmacological and molecular inhibition reveals that these mechanisms are dependent on the adaptor proteins AP-1 and AP-3. Further, shRNA-mediated knockdown reveals differential roles for AP-1 and AP-3 in VGLUT2 recycling. PMID:29123471

VGLUT2 Trafficking Is Differentially Regulated by Adaptor Proteins AP-1 and AP-3.

PubMed

Li, Haiyan; Santos, Magda S; Park, Chihyung K; Dobry, Yuriy; Voglmaier, Susan M

2017-01-01

Release of the major excitatory neurotransmitter glutamate by synaptic vesicle exocytosis depends on glutamate loading into synaptic vesicles by vesicular glutamate transporters (VGLUTs). The two principal isoforms, VGLUT1 and 2, exhibit a complementary pattern of expression in adult brain that broadly distinguishes cortical (VGLUT1) and subcortical (VGLUT2) systems, and correlates with distinct physiological properties in synapses expressing these isoforms. Differential trafficking of VGLUT1 and 2 has been suggested to underlie their functional diversity. Increasing evidence suggests individual synaptic vesicle proteins use specific sorting signals to engage specialized biochemical mechanisms to regulate their recycling. We observed that VGLUT2 recycles differently in response to high frequency stimulation than VGLUT1. Here we further explore the trafficking of VGLUT2 using a pHluorin-based reporter, VGLUT2-pH. VGLUT2-pH exhibits slower rates of both exocytosis and endocytosis than VGLUT1-pH. VGLUT2-pH recycling is slower than VGLUT1-pH in both hippocampal neurons, which endogenously express mostly VGLUT1, and thalamic neurons, which endogenously express mostly VGLUT2, indicating that protein identity, not synaptic vesicle membrane or neuronal cell type, controls sorting. We characterize sorting signals in the C-terminal dileucine-like motif, which plays a crucial role in VGLUT2 trafficking. Disruption of this motif abolishes synaptic targeting of VGLUT2 and essentially eliminates endocytosis of the transporter. Mutational and biochemical analysis demonstrates that clathrin adaptor proteins (APs) interact with VGLUT2 at the dileucine-like motif. VGLUT2 interacts with AP-2, a well-studied adaptor protein for clathrin mediated endocytosis. In addition, VGLUT2 also interacts with the alternate adaptors, AP-1 and AP-3. VGLUT2 relies on distinct recycling mechanisms from VGLUT1. Abrogation of these differences by pharmacological and molecular inhibition reveals that these mechanisms are dependent on the adaptor proteins AP-1 and AP-3. Further, shRNA-mediated knockdown reveals differential roles for AP-1 and AP-3 in VGLUT2 recycling.

Development of a novel cell sorting method that samples population diversity in flow cytometry.

PubMed

Osborne, Geoffrey W; Andersen, Stacey B; Battye, Francis L

2015-11-01

Flow cytometry based electrostatic cell sorting is an important tool in the separation of cell populations. Existing instruments can sort single cells into multi-well collection plates, and keep track of cell of origin and sorted well location. However currently single sorted cell results reflect the population distribution and fail to capture the population diversity. Software was designed that implements a novel sorting approach, "Slice and Dice Sorting," that links a graphical representation of a multi-well plate to logic that ensures that single cells are sampled and sorted from all areas defined by the sort region/s. Therefore the diversity of the total population is captured, and the more frequently occurring or rarer cell types are all sampled. The sorting approach was tested computationally, and using functional cell based assays. Computationally we demonstrate that conventional single cell sorting can sample as little as 50% of the population diversity dependant on the population distribution, and that Slice and Dice sorting samples much more of the variety present within a cell population. We then show by sorting single cells into wells using the Slice and Dice sorting method that there are cells sorted using this method that would be either rarely sorted, or not sorted at all using conventional single cell sorting approaches. The present study demonstrates a novel single cell sorting method that samples much more of the population diversity than current methods. It has implications in clonal selection, stem cell sorting, single cell sequencing and any areas where population heterogeneity is of importance. © 2015 International Society for Advancement of Cytometry.

Advancing the Fork detector for quantitative spent nuclear fuel verification

DOE PAGES

Vaccaro, S.; Gauld, I. C.; Hu, J.; ...

2018-01-31

The Fork detector is widely used by the safeguards inspectorate of the European Atomic Energy Community (EURATOM) and the International Atomic Energy Agency (IAEA) to verify spent nuclear fuel. Fork measurements are routinely performed for safeguards prior to dry storage cask loading. Additionally, spent fuel verification will be required at the facilities where encapsulation is performed for acceptance in the final repositories planned in Sweden and Finland. The use of the Fork detector as a quantitative instrument has not been prevalent due to the complexity of correlating the measured neutron and gamma ray signals with fuel inventories and operator declarations.more » A spent fuel data analysis module based on the ORIGEN burnup code was recently implemented to provide automated real-time analysis of Fork detector data. This module allows quantitative predictions of expected neutron count rates and gamma units as measured by the Fork detectors using safeguards declarations and available reactor operating data. This study describes field testing of the Fork data analysis module using data acquired from 339 assemblies measured during routine dry cask loading inspection campaigns in Europe. Assemblies include both uranium oxide and mixed-oxide fuel assemblies. More recent measurements of 50 spent fuel assemblies at the Swedish Central Interim Storage Facility for Spent Nuclear Fuel are also analyzed. An evaluation of uncertainties in the Fork measurement data is performed to quantify the ability of the data analysis module to verify operator declarations and to develop quantitative go/no-go criteria for safeguards verification measurements during cask loading or encapsulation operations. The goal of this approach is to provide safeguards inspectors with reliable real-time data analysis tools to rapidly identify discrepancies in operator declarations and to detect potential partial defects in spent fuel assemblies with improved reliability and minimal false positive alarms. Finally, the results are summarized, and sources and magnitudes of uncertainties are identified, and the impact of analysis uncertainties on the ability to confirm operator declarations is quantified.« less

Advancing the Fork detector for quantitative spent nuclear fuel verification

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vaccaro, S.; Gauld, I. C.; Hu, J.

The Fork detector is widely used by the safeguards inspectorate of the European Atomic Energy Community (EURATOM) and the International Atomic Energy Agency (IAEA) to verify spent nuclear fuel. Fork measurements are routinely performed for safeguards prior to dry storage cask loading. Additionally, spent fuel verification will be required at the facilities where encapsulation is performed for acceptance in the final repositories planned in Sweden and Finland. The use of the Fork detector as a quantitative instrument has not been prevalent due to the complexity of correlating the measured neutron and gamma ray signals with fuel inventories and operator declarations.more » A spent fuel data analysis module based on the ORIGEN burnup code was recently implemented to provide automated real-time analysis of Fork detector data. This module allows quantitative predictions of expected neutron count rates and gamma units as measured by the Fork detectors using safeguards declarations and available reactor operating data. This study describes field testing of the Fork data analysis module using data acquired from 339 assemblies measured during routine dry cask loading inspection campaigns in Europe. Assemblies include both uranium oxide and mixed-oxide fuel assemblies. More recent measurements of 50 spent fuel assemblies at the Swedish Central Interim Storage Facility for Spent Nuclear Fuel are also analyzed. An evaluation of uncertainties in the Fork measurement data is performed to quantify the ability of the data analysis module to verify operator declarations and to develop quantitative go/no-go criteria for safeguards verification measurements during cask loading or encapsulation operations. The goal of this approach is to provide safeguards inspectors with reliable real-time data analysis tools to rapidly identify discrepancies in operator declarations and to detect potential partial defects in spent fuel assemblies with improved reliability and minimal false positive alarms. Finally, the results are summarized, and sources and magnitudes of uncertainties are identified, and the impact of analysis uncertainties on the ability to confirm operator declarations is quantified.« less

Plasma Membrane Ca2+-ATPase 4 in Murine Epididymis: Secretion of Splice Variants in the Luminal Fluid and a Role in Sperm Maturation1

PubMed Central

Patel, Ramkrishna; Al-Dossary, Amal A.; Stabley, Deborah L.; Barone, Carol; Galileo, Deni S.; Strehler, Emanuel E.; Martin-DeLeon, Patricia A.

2013-01-01

ABSTRACT Plasma membrane Ca2+-ATPase isoform 4 (PMCA4) is the primary Ca2+ efflux pump in murine sperm, where it regulates motility. In Pmca4 null sperm, motility loss results in infertility. We have shown that murine sperm PMCA4b interacts with Ca2+/CaM-dependent serine kinase (CASK) in regulating Ca2+ homeostasis and motility. However, recent work indicated that the bovine PMCA4a splice variant (missing in testis) is epididymally expressed, along with 4b, and may be transferred to sperm. Here we show, via conventional and in situ RT-PCR, that both the splice variants of Pmca4 mRNA are expressed in murine testis and throughout the epididymis. Immunofluorescence localized PMCA4a to the apical membrane of the epididymal epithelium, and Western analysis not only confirmed its presence but showed for the first time that PMCA4a and PMCA4b are secreted in the epididymal luminal fluid (ELF), from which epididymosomes containing PMCA4a were isolated. Flow cytometry indicated the presence of PMCA4a on mature caudal sperm where it was increased ∼5-fold compared to caput sperm (detected by Western blotting) and ∼2-fold after incubation in ELF, revealing in vitro uptake and implicating PMCA4a in epididymal sperm maturation. Coimmunoprecipitation using pan-PMCA4 antibodies, revealed that both variants associate with CASK, suggesting their presence in a complex. Because they have different kinetic properties for Ca2+ transport and different abilities to bind to CASK, our study suggests a mechanism for combining the functional attributes of both PMCA4 variants, leading to heightened efficiency of the pump in the maintenance of Ca2+ homeostasis, which is crucial for normal motility and male fertility. PMID:23699388

Evaluation of Aluminum-Boron Carbide Neutron Absorbing Materials for Interim Storage of Used Nuclear Fuel

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Lumin; Wierschke, Jonathan Brett

2015-04-08

The objective of this work was to understand the corrosion behavior of Boral® and Bortec® neutron absorbers over long-term deployment in a used nuclear fuel dry cask storage environment. Corrosion effects were accelerated by flowing humidified argon through an autoclave at temperatures up to 570°C. Test results show little corrosion of the aluminum matrix but that boron is leaching out of the samples. Initial tests performed at 400 and 570°C were hampered by reduced flow caused by the rapid build-up of solid deposits in the outlet lines. Analysis of the deposits by XRD shows that the deposits are comprised ofmore » boron trioxide and sassolite (H 3BO 3). The collection of boron- containing compounds in the outlet lines indicated that boron was being released from the samples. Observation of the exposed samples using SEM and optical microscopy show the growth of new phases in the samples. These phases were most prominent in Bortec® samples exposed at 570°C. Samples of Boral® exposed at 570°C showed minimal new phase formation but showed nearly the complete loss of boron carbide particles. Boron carbide loss was also significant in Boral samples at 400°C. However, at 400°C phases similar to those found in Bortec® were observed. The rapid loss of the boron carbide particles in the Boral® is suspected to inhibit the formation of the new secondary phases. However, Material samples in an actual dry cask environment would be exposed to temperatures closer to 300°C and less water than the lowest test. The results from this study conclude that at the temperature and humidity levels present in a dry cask environment, corrosion and boron leaching will have no effect on the performance of Boral® and Bortec® to maintain criticality control.« less

Depleted uranium hexafluoride: The source material for advanced shielding systems

DOE Office of Scientific and Technical Information (OSTI.GOV)

Quapp, W.J.; Lessing, P.A.; Cooley, C.R.

1997-02-01

The U.S. Department of Energy (DOE) has a management challenge and financial liability problem in the form of 50,000 cylinders containing 555,000 metric tons of depleted uranium hexafluoride (UF{sub 6}) that are stored at the gaseous diffusion plants. DOE is evaluating several options for the disposition of this UF{sub 6}, including continued storage, disposal, and recycle into a product. Based on studies conducted to date, the most feasible recycle option for the depleted uranium is shielding in low-level waste, spent nuclear fuel, or vitrified high-level waste containers. Estimates for the cost of disposal, using existing technologies, range between $3.8 andmore » $11.3 billion depending on factors such as the disposal site and the applicability of the Resource Conservation and Recovery Act (RCRA). Advanced technologies can reduce these costs, but UF{sub 6} disposal still represents large future costs. This paper describes an application for depleted uranium in which depleted uranium hexafluoride is converted into an oxide and then into a heavy aggregate. The heavy uranium aggregate is combined with conventional concrete materials to form an ultra high density concrete, DUCRETE, weighing more than 400 lb/ft{sup 3}. DUCRETE can be used as shielding in spent nuclear fuel/high-level waste casks at a cost comparable to the lower of the disposal cost estimates. Consequently, the case can be made that DUCRETE shielded casks are an alternative to disposal. In this case, a beneficial long term solution is attained for much less than the combined cost of independently providing shielded casks and disposing of the depleted uranium. Furthermore, if disposal is avoided, the political problems associated with selection of a disposal location are also avoided. Other studies have also shown cost benefits for low level waste shielded disposal containers.« less

Bias estimates used in lieu of validation of fission products and minor actinides in MCNP K eff calculations for PWR burnup credit casks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mueller, Don E.; Marshall, William J.; Wagner, John C.

The U.S. Nuclear Regulatory Commission (NRC) Division of Spent Fuel Storage and Transportation recently issued Interim Staff Guidance (ISG) 8, Revision 3. This ISG provides guidance for burnup credit (BUC) analyses supporting transport and storage of PWR pressurized water reactor (PWR) fuel in casks. Revision 3 includes guidance for addressing validation of criticality (k eff) calculations crediting the presence of a limited set of fission products and minor actinides (FP&MA). Based on previous work documented in NUREG/CR-7109, recommendation 4 of ISG-8, Rev. 3, includes a recommendation to use 1.5 or 3% of the FP&MA worth to conservatively cover the biasmore » due to the specified FP&MAs. This bias is supplementary to the bias and bias uncertainty resulting from validation of k eff calculations for the major actinides in SNF and does not address extension to actinides and fission products beyond those identified herein. The work described in this report involves comparison of FP&MA worths calculated using SCALE and MCNP with ENDF/B-V, -VI, and -VII based nuclear data and supports use of the 1.5% FP&MA worth bias when either SCALE or MCNP codes are used for criticality calculations, provided the other conditions of the recommendation 4 are met. The method used in this report may also be applied to demonstrate the applicability of the 1.5% FP&MA worth bias to other codes using ENDF/B V, VI or VII based nuclear data. The method involves use of the applicant s computational method to generate FP&MA worths for a reference SNF cask model using specified spent fuel compositions. The applicant s FP&MA worths are then compared to reference values provided in this report. The applicants FP&MA worths should not exceed the reference results by more than 1.5% of the reference FP&MA worths.« less

Advancing the Fork detector for quantitative spent nuclear fuel verification

NASA Astrophysics Data System (ADS)

Vaccaro, S.; Gauld, I. C.; Hu, J.; De Baere, P.; Peterson, J.; Schwalbach, P.; Smejkal, A.; Tomanin, A.; Sjöland, A.; Tobin, S.; Wiarda, D.

2018-04-01

The Fork detector is widely used by the safeguards inspectorate of the European Atomic Energy Community (EURATOM) and the International Atomic Energy Agency (IAEA) to verify spent nuclear fuel. Fork measurements are routinely performed for safeguards prior to dry storage cask loading. Additionally, spent fuel verification will be required at the facilities where encapsulation is performed for acceptance in the final repositories planned in Sweden and Finland. The use of the Fork detector as a quantitative instrument has not been prevalent due to the complexity of correlating the measured neutron and gamma ray signals with fuel inventories and operator declarations. A spent fuel data analysis module based on the ORIGEN burnup code was recently implemented to provide automated real-time analysis of Fork detector data. This module allows quantitative predictions of expected neutron count rates and gamma units as measured by the Fork detectors using safeguards declarations and available reactor operating data. This paper describes field testing of the Fork data analysis module using data acquired from 339 assemblies measured during routine dry cask loading inspection campaigns in Europe. Assemblies include both uranium oxide and mixed-oxide fuel assemblies. More recent measurements of 50 spent fuel assemblies at the Swedish Central Interim Storage Facility for Spent Nuclear Fuel are also analyzed. An evaluation of uncertainties in the Fork measurement data is performed to quantify the ability of the data analysis module to verify operator declarations and to develop quantitative go/no-go criteria for safeguards verification measurements during cask loading or encapsulation operations. The goal of this approach is to provide safeguards inspectors with reliable real-time data analysis tools to rapidly identify discrepancies in operator declarations and to detect potential partial defects in spent fuel assemblies with improved reliability and minimal false positive alarms. The results are summarized, and sources and magnitudes of uncertainties are identified, and the impact of analysis uncertainties on the ability to confirm operator declarations is quantified.

Plasma membrane Ca2+-ATPase 4 in murine epididymis: secretion of splice variants in the luminal fluid and a role in sperm maturation.

PubMed

Patel, Ramkrishna; Al-Dossary, Amal A; Stabley, Deborah L; Barone, Carol; Galileo, Deni S; Strehler, Emanuel E; Martin-DeLeon, Patricia A

2013-07-01

Plasma membrane Ca(2+)-ATPase isoform 4 (PMCA4) is the primary Ca(2+) efflux pump in murine sperm, where it regulates motility. In Pmca4 null sperm, motility loss results in infertility. We have shown that murine sperm PMCA4b interacts with Ca(2+)/CaM-dependent serine kinase (CASK) in regulating Ca(2+) homeostasis and motility. However, recent work indicated that the bovine PMCA4a splice variant (missing in testis) is epididymally expressed, along with 4b, and may be transferred to sperm. Here we show, via conventional and in situ RT-PCR, that both the splice variants of Pmca4 mRNA are expressed in murine testis and throughout the epididymis. Immunofluorescence localized PMCA4a to the apical membrane of the epididymal epithelium, and Western analysis not only confirmed its presence but showed for the first time that PMCA4a and PMCA4b are secreted in the epididymal luminal fluid (ELF), from which epididymosomes containing PMCA4a were isolated. Flow cytometry indicated the presence of PMCA4a on mature caudal sperm where it was increased ~5-fold compared to caput sperm (detected by Western blotting) and ~2-fold after incubation in ELF, revealing in vitro uptake and implicating PMCA4a in epididymal sperm maturation. Coimmunoprecipitation using pan-PMCA4 antibodies, revealed that both variants associate with CASK, suggesting their presence in a complex. Because they have different kinetic properties for Ca(2+) transport and different abilities to bind to CASK, our study suggests a mechanism for combining the functional attributes of both PMCA4 variants, leading to heightened efficiency of the pump in the maintenance of Ca(2+) homeostasis, which is crucial for normal motility and male fertility.

Parallel sort with a ranged, partitioned key-value store in a high perfomance computing environment

DOEpatents

Bent, John M.; Faibish, Sorin; Grider, Gary; Torres, Aaron; Poole, Stephen W.

2016-01-26

Improved sorting techniques are provided that perform a parallel sort using a ranged, partitioned key-value store in a high performance computing (HPC) environment. A plurality of input data files comprising unsorted key-value data in a partitioned key-value store are sorted. The partitioned key-value store comprises a range server for each of a plurality of ranges. Each input data file has an associated reader thread. Each reader thread reads the unsorted key-value data in the corresponding input data file and performs a local sort of the unsorted key-value data to generate sorted key-value data. A plurality of sorted, ranged subsets of each of the sorted key-value data are generated based on the plurality of ranges. Each sorted, ranged subset corresponds to a given one of the ranges and is provided to one of the range servers corresponding to the range of the sorted, ranged subset. Each range server sorts the received sorted, ranged subsets and provides a sorted range. A plurality of the sorted ranges are concatenated to obtain a globally sorted result.

Flammable and noxious gas sensing using a microtripolar electrode sensor with diameter and chirality sorted single-walled carbon nanotubes

NASA Astrophysics Data System (ADS)

Cai, Shengbing; Duan, Zhe min; Zhang, Yong

2013-08-01

We report on the utilization of densely packed (˜10 SWCNTs µm-1), well-aligned arrays of single-chirality single-walled carbon nanotubes (SWCNTs) as an effective thin-film for integration into a gas sensor with a microtripolar electrode, based on field ionization by dielectrophoretic assembly from a monodisperse SWCNTs solution obtained by polymer-mediated sorting. The sensor is characterized as a field ionization electrode with sorted SWCNTs acting as both the sensing material and transducer gas concentrated directly into an electrical signal, an extractor serving to improve electric field uniformity and a collector electrode completing the current path. The gas sensing properties toward flammable and noxious gases, such as CO and H2, were investigated at room temperature. Besides the high sensitivity, the as-fabricated sensor exhibited attractive behaviors in terms of both the detection limit and a fast response, suggesting that our sensor could be used to partly circumvent the low sensing selectivity, long recovery time or irreversibility and allow for a preferential identification of the selected flammable and noxious analytes. Interestingly, the excellent sensing behaviors of the sensors based on the field ionization effect derive directly from the combined effects of the high-quality, low defect SWCNTs arrays, which leads to a small device-to-device variation in the properties and the optimization of electrode fabrication, highlighting the sensor as an appealing candidate in view of nanotube electronics.

N-linked oligosaccharides on the low density lipoprotein receptor homolog SorLA/LR11 are modified with terminal GalNAc-4-SO4 in kidney and brain.

PubMed

Fiete, Dorothy; Mi, Yiling; Oats, Edward L; Beranek, Mary C; Baenziger, Jacques U

2007-01-19

Sorting protein-related receptor (SorLA/LR11) is a highly conserved mosaic receptor that is expressed by cells in a number of different tissues including principal cells of the collecting ducts in the kidney and neurons in the central and peripheral nervous systems. SorLA/LR11 has features that indicate it serves as a sorting receptor shuttling between the plasma membrane, endosomes, and the Golgi. We have found that a fraction of SorLA/LR11 that is synthesized in the kidney and the brain bears N-linked oligosaccharides that are modified with terminal beta1,4-linked GalNAc-4-SO(4). Oligosaccharides located in the vacuolar sorting (Vps) 10p domain (Vps10p domain) are modified with beta1,4-linked GalNAc when the Vps10p domain is expressed in cells along with either of two recently cloned protein-specific beta1,4GalNAc-transferases, GalNAcTIII and GalNAcTIV. Either of two sequences with basic amino acids located within the Vps10p domain is able to mediate recognition by these beta1,4GalNAc-transferases. The highly specific modification of oligosaccharides in the Vps10p domain of SorLA/LR11 with terminal GalNAc-4-SO(4) suggests that this unusual modification may modulate the interaction of SorLA/LR11 with proteins and influence their trafficking.

Phosphorylation-dependent trafficking of plasma membrane proteins in animal and plant cells.

PubMed

Offringa, Remko; Huang, Fang

2013-09-01

In both unicellular and multicellular organisms, transmembrane (TM) proteins are sorted to and retained at specific membrane domains by endomembrane trafficking mechanisms that recognize sorting signals in the these proteins. The trafficking and distribution of plasma membrane (PM)-localized TM proteins (PM proteins), especially of those PM proteins that show an asymmetric distribution over the PM, has received much attention, as their proper PM localization is crucial for elementary signaling and transport processes, and defects in their localization often lead to severe disease symptoms or developmental defects. The subcellular localization of PM proteins is dynamically regulated by post-translational modifications, such as phosphorylation and ubiquitination. These modificaitons mostly occur on sorting signals that are located in the larger cytosolic domains of the cargo proteins. Here we review the effects of phosphorylation of PM proteins on their trafficking, and present the key examples from the animal field that have been subject to studies for already several decades, such as that of aquaporin 2 and the epidermal growth factor receptor. Our knowledge on cargo trafficking in plants is largely based on studies of the family of PIN FORMED (PIN) carriers that mediate the efflux of the plant hormone auxin. We will review what is known on the subcellular distribution and trafficking of PIN proteins, with a focus on how this is modulated by phosphorylation, and identify and discuss analogies and differences in trafficking with the well-studied animal examples. © 2013 Institute of Botany, Chinese Academy of Sciences.

Functional Characterization of Ice Plant SKD1, an AAA-Type ATPase Associated with the Endoplasmic Reticulum-Golgi Network, and Its Role in Adaptation to Salt Stress1[W

PubMed Central

Jou, Yingtzy; Chiang, Chih-Pin; Jauh, Guang-Yuh; Yen, Hungchen Emilie

2006-01-01

A salt-induced gene mcSKD1 (suppressor of K+ transport growth defect) able to facilitate K+ uptake has previously been identified from the halophyte ice plant (Mesembryanthemum crystallinum). The sequence of mcSKD1 is homologous to vacuolar protein sorting 4, an ATPase associated with a variety of cellular activities-type ATPase that participates in the sorting of vacuolar proteins into multivesicular bodies in yeast (Saccharomyces cerevisiae). Recombinant mcSKD1 exhibited ATP hydrolytic activities in vitro with a half-maximal rate at an ATP concentration of 1.25 mm. Point mutations on active site residues abolished its ATPase activity. ADP is both a product and a strong inhibitor of the reaction. ADP-binding form of mcSDK1 greatly reduced its catalytic activity. The mcSKD1 protein accumulated ubiquitously in both vegetative and reproductive parts of plants. Highest accumulation was observed in cells actively engaging in the secretory processes, such as bladder cells of leaf epidermis. Membrane fractionation and double-labeling immunofluorescence showed the predominant localization of mcSKD1 in the endoplasmic reticulum-Golgi network. Immunoelectron microscopy identified the formation of mcSKD1 proteins into small aggregates in the cytosol and associated with membrane continuum within the endomembrane compartments. These results indicated that this ATPase participates in the endoplasmic reticulum-Golgi mediated protein sorting machinery for both housekeeping function and compartmentalization of excess Na+ under high salinity. PMID:16581876

Assessment of FUN-1 vital dye staining: Yeast with a block in the vacuolar sorting pathway have impaired ability to form CIVS when stained with FUN-1 fluorescent dye.

PubMed

Essary, Brandin D; Marshall, Pamela A

2009-08-01

FUN-1 [2-chloro-4-(2,3-dihydro-3-methyl-(benzo-1,3-thiazol-2-yl)-methylidene)-1-phenylquinolinium iodide] is a fluorescent dye used in studies of yeast and other fungi to monitor cell viability in the research lab and to assay for active fungal infection in the clinical setting. When the plasma membrane is intact, fungal cells internalize FUN-1 and the dye is seen as diffuse green cytosolic fluorescence. FUN-1 is then transported to the vacuole in metabolically active wild type cells and subsequently is compacted into fluorescent red cylindrical intravacuolar structures (CIVS) by an unknown transport pathway. This dye is used to determine yeast viability, as only live cells form CIVS. However, in live Saccharomyces cerevisiae with impaired protein sorting to the yeast vacuole, we report decreased to no CIVS formation, depending on the cellular location of the block in the sorting pathway. Cells with a block in vesicle-mediated transport from the Golgi to prevacuolar compartment (PVC) or with a block in recycling from the PVC to the Golgi demonstrate a substantial impairment in CIVS formation. Instead, the FUN-1 dye is seen either in small punctate structures under fluorescence or as diffuse red cytosol under white light. Thus, researchers using FUN-1 should be cognizant of the limitations of this procedure in determining cell viability as there are viable yeast mutants with impaired CIVS formation.

Evidence for a role of SNX16 in regulating traffic between the early and later endosomal compartments.

PubMed

Hanson, Brendon J; Hong, Wanjin

2003-09-05

Sorting nexins (SNXs) are a growing family of proteins characterized by the presence of a PX domain. The PX domain mediates membrane association by interaction with phosphoinositides. The SNXs are generally believed to participate in membrane trafficking, but information regarding the function of individual proteins is limited. In this report, we describe the major characteristics of one member, SNX16. SNX16 is a novel 343-amino acid protein consisting of a central PX domain followed by a potential coiled-coil domain and a C-terminal region. Like other sorting nexins, SNX16 associates with the membrane via the PX domain which interacts with the phospholipid phosphatidylinositol 3-phosphate. We show via biochemical and cellular studies that SNX16 is distributed in both early and late endosome/lysosome structures. The coiled-coil domain is necessary for localization to the later endosomal structures, as mutant SNX16 lacking this domain was found only in early endosomes. Trafficking of internalized epidermal growth factor was also delayed by this SNX16 mutant, as these cells showed a delay in the segregation of epidermal growth factor in the early endosome for its delivery to later compartments. In addition, the coiled-coil domain is shown here to be important for homo-oligomerization of SNX16. Taken together, these results suggest that SNX16 is a sorting nexin that may function in the trafficking of proteins between the early and late endosomal compartments.

Dynamin-like protein 1 at the Golgi complex: A novel component of the sorting/targeting machinery en route to the plasma membrane

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bonekamp, Nina A.; Vormund, Kerstin; Jacob, Ralf

2010-12-10

The final step in the liberation of secretory vesicles from the trans-Golgi network (TGN) involves the mechanical action of the large GTPase dynamin as well as conserved dynamin-independent fission mechanisms, e.g. mediated by Brefeldin A-dependent ADP-ribosylated substrate (BARS). Another member of the dynamin family is the mammalian dynamin-like protein 1 (DLP1/Drp1) that is known to constrict and tubulate membranes, and to divide mitochondria and peroxisomes. Here, we examined a potential role for DLP1 at the Golgi complex. DLP1 localized to the Golgi complex in some but not all cell lines tested, thus explaining controversial reports on its cellular distribution. Aftermore » silencing of DLP1, an accumulation of the apical reporter protein YFP-GL-GPI, but not the basolateral reporter VSVG-SP-GFP at the Golgi complex was observed. A reduction in the transport of YFP-GL-GPI to the plasma membrane was confirmed by surface immunoprecipitation and TGN-exit assays. In contrast, YFP-GL-GPI trafficking was not disturbed in cells silenced for BARS, which is involved in basolateral sorting and trafficking of VSVG-SP-GFP in COS-7 cells. Our data indicate a new role for DLP1 at the Golgi complex and thus a role for DLP1 as a novel component of the apical sorting machinery at the TGN is discussed.« less

Developing models of how cognitive improvements change functioning: Mediation, moderation and moderated mediation

PubMed Central

Wykes, Til; Reeder, Clare; Huddy, Vyv; Taylor, Rumina; Wood, Helen; Ghirasim, Natalia; Kontis, Dimitrios; Landau, Sabine

2012-01-01

Background Cognitive remediation (CRT) affects functioning but the extent and type of cognitive improvements necessary are unknown. Aim To develop and test models of how cognitive improvement transfers to work behaviour using the data from a current service. Method Participants (N49) with a support worker and a paid or voluntary job were offered CRT in a Phase 2 single group design with three assessments: baseline, post therapy and follow-up. Working memory, cognitive flexibility, planning and work outcomes were assessed. Results Three models were tested (mediation — cognitive improvements drive functioning improvement; moderation — post treatment cognitive level affects the impact of CRT on functioning; moderated mediation — cognition drives functioning improvements only after a certain level is achieved). There was evidence of mediation (planning improvement associated with improved work quality). There was no evidence that cognitive flexibility (total Wisconsin Card Sorting Test errors) and working memory (Wechsler Adult Intelligence Scale III digit span) mediated work functioning despite significant effects. There was some evidence of moderated mediation for planning improvement if participants had poorer memory and/or made fewer WCST errors. The total CRT effect on work quality was d = 0.55, but the indirect (planning-mediated CRT effect) was d = 0.082 Conclusion Planning improvements led to better work quality but only accounted for a small proportion of the total effect on work outcome. Other specific and non-specific effects of CRT and the work programme are likely to account for some of the remaining effect. This is the first time complex models have been tested and future Phase 3 studies need to further test mediation and moderated mediation models. PMID:22503640

Small extracellular vesicles secreted from senescent cells promote cancer cell proliferation through EphA2

PubMed Central

Takasugi, Masaki; Okada, Ryo; Takahashi, Akiko; Virya Chen, David; Watanabe, Sugiko; Hara, Eiji

2017-01-01

Cellular senescence prevents the proliferation of cells at risk for neoplastic transformation. However, the altered secretome of senescent cells can promote the growth of the surrounding cancer cells. Although extracellular vesicles (EVs) have emerged as new players in intercellular communication, their role in the function of senescent cell secretome has been largely unexplored. Here, we show that exosome-like small EVs (sEVs) are important mediators of the pro-tumorigenic function of senescent cells. sEV-associated EphA2 secreted from senescent cells binds to ephrin-A1, that is, highly expressed in several types of cancer cells and promotes cell proliferation through EphA2/ephrin-A1 reverse signalling. sEV sorting of EphA2 is increased in senescent cells because of its enhanced phosphorylation resulting from oxidative inactivation of PTP1B phosphatase. Our results demonstrate a novel mechanism of reactive oxygen species (ROS)-regulated cargo sorting into sEVs, which is critical for the potentially deleterious growth-promoting effect of the senescent cell secretome. PMID:28585531

The VirusBanker database uses a Java program to allow flexible searching through Bunyaviridae sequences.

PubMed

Fourment, Mathieu; Gibbs, Mark J

2008-02-05

Viruses of the Bunyaviridae have segmented negative-stranded RNA genomes and several of them cause significant disease. Many partial sequences have been obtained from the segments so that GenBank searches give complex results. Sequence databases usually use HTML pages to mediate remote sorting, but this approach can be limiting and may discourage a user from exploring a database. The VirusBanker database contains Bunyaviridae sequences and alignments and is presented as two spreadsheets generated by a Java program that interacts with a MySQL database on a server. Sequences are displayed in rows and may be sorted using information that is displayed in columns and includes data relating to the segment, gene, protein, species, strain, sequence length, terminal sequence and date and country of isolation. Bunyaviridae sequences and alignments may be downloaded from the second spreadsheet with titles defined by the user from the columns, or viewed when passed directly to the sequence editor, Jalview. VirusBanker allows large datasets of aligned nucleotide and protein sequences from the Bunyaviridae to be compiled and winnowed rapidly using criteria that are formulated heuristically.

CCC- and WASH-mediated endosomal sorting of LDLR is required for normal clearance of circulating LDL

PubMed Central

Bartuzi, Paulina; Billadeau, Daniel D.; Favier, Robert; Rong, Shunxing; Dekker, Daphne; Fedoseienko, Alina; Fieten, Hille; Wijers, Melinde; Levels, Johannes H.; Huijkman, Nicolette; Kloosterhuis, Niels; van der Molen, Henk; Brufau, Gemma; Groen, Albert K.; Elliott, Alison M.; Kuivenhoven, Jan Albert; Plecko, Barbara; Grangl, Gernot; McGaughran, Julie; Horton, Jay D.; Burstein, Ezra; Hofker, Marten H.; van de Sluis, Bart

2016-01-01

The low-density lipoprotein receptor (LDLR) plays a pivotal role in clearing atherogenic circulating low-density lipoprotein (LDL) cholesterol. Here we show that the COMMD/CCDC22/CCDC93 (CCC) and the Wiskott–Aldrich syndrome protein and SCAR homologue (WASH) complexes are both crucial for endosomal sorting of LDLR and for its function. We find that patients with X-linked intellectual disability caused by mutations in CCDC22 are hypercholesterolaemic, and that COMMD1-deficient dogs and liver-specific Commd1 knockout mice have elevated plasma LDL cholesterol levels. Furthermore, Commd1 depletion results in mislocalization of LDLR, accompanied by decreased LDL uptake. Increased total plasma cholesterol levels are also seen in hepatic COMMD9-deficient mice. Inactivation of the CCC-associated WASH complex causes LDLR mislocalization, increased lysosomal degradation of LDLR and impaired LDL uptake. Furthermore, a mutation in the WASH component KIAA0196 (strumpellin) is associated with hypercholesterolaemia in humans. Altogether, this study provides valuable insights into the mechanisms regulating cholesterol homeostasis and LDLR trafficking. PMID:26965651

GGA3 Functions as a Switch to Promote Met Receptor Recycling, Essential for Sustained ERK and Cell Migration

PubMed Central

Parachoniak, Christine Anna; Luo, Yi; Abella, Jasmine Vanessa; Keen, James H.; Park, Morag

2011-01-01

Summary Cells are dependent on correct sorting of activated receptor tyrosine kinases (RTKs) for the outcome of growth factor signaling. Upon activation, RTKs are coupled through the endocytic machinery for degradation, or recycled to the cell surface. However, the molecular mechanisms governing RTK recycling are poorly understood. Here, we show that Golgi-localized gamma-ear containing Arf-binding protein 3 (GGA3) interacts selectively with the Met/Hepatocyte Growth Factor RTK when stimulated, to sort it for recycling in association with “gyrating”-clathrin. GGA3 loss abrogates Met recycling from a Rab4 endosomal subdomain, resulting in pronounced trafficking of Met towards degradation. Decreased Met recycling attenuates ERK activation and cell migration. Met recycling, sustained ERK activation and migration require interaction of GGA3 with Arf6 and an unexpected association with the Crk adaptor. The data show that GGA3 defines an active recycling pathway and support a broader role for GGA3-mediated cargo selection in targeting receptors destined for recycling. PMID:21664574

Targeting sortilin in immune cells reduces proinflammatory cytokines and atherosclerosis

PubMed Central

Mortensen, Martin B.; Kjolby, Mads; Gunnersen, Stine; Larsen, Jakob V.; Palmfeldt, Johan; Falk, Erling; Nykjaer, Anders; Bentzon, Jacob F.

2014-01-01

Genome-wide association studies have identified a link between genetic variation at the human chromosomal locus 1p13.3 and coronary artery disease. The gene encoding sortilin (SORT1) has been implicated as the causative gene within the locus, as sortilin regulates hepatic lipoprotein metabolism. Here we demonstrated that sortilin also directly affects atherogenesis, independent of its regulatory role in lipoprotein metabolism. In a mouse model of atherosclerosis, deletion of Sort1 did not alter plasma cholesterol levels, but reduced the development of both early and late atherosclerotic lesions. We determined that sortilin is a high-affinity receptor for the proinflammatory cytokines IL-6 and IFN-γ. Moreover, macrophages and Th1 cells (both of which mediate atherosclerotic plaque formation) lacking sortilin had reduced secretion of IL-6 and IFN-γ, but not of other measured cytokines. Transfer of sortilin-deficient BM into irradiated atherosclerotic mice reduced atherosclerosis and systemic markers of inflammation. Together, these data demonstrate that sortilin influences cytokine secretion and that targeting sortilin in immune cells attenuates inflammation and reduces atherosclerosis. PMID:25401472

27 CFR 31.231 - Destruction of marks and brands on wine containers.

Code of Federal Regulations, 2010 CFR

2010-04-01

... brands on wine containers. 31.231 Section 31.231 Alcohol, Tobacco Products and Firearms ALCOHOL AND... § 31.231 Destruction of marks and brands on wine containers. A dealer who empties any cask, barrel, keg, or other bulk container of wine must scrape or obliterate from the empty container all marks, brands...

PATRAM '80. Proceedings. Volume 2

DOE Office of Scientific and Technical Information (OSTI.GOV)

Huebner, H.W.

1980-01-01

Volume 2 contains papers from the following sessions: Safeguards-Related Problems; Neutronics and Criticality; Operations and Systems Experience II; Plutonium Systems; Intermediate Storage in Casks; Operations and Systems Planning; Institutional Issues; Structural and Thermal Evaluation I; Poster Session B; Extended Testing I; Structural and Thermal Evaluation II; Extended Testing II; and Emergency Preparedness and Response. Individual papers were processed. (LM)

ROUGHING IT. THE OLD MAN AND THE SEA. SHORT STORIES. POEMS. LITERATURE CURRICULUM III, TEACHER VERSION.

ERIC Educational Resources Information Center

KITZHABER, ALBERT R.

A TEACHER VERSION OF A LITERATURE CURRICULUM GUIDE WAS PROVIDED FOR TWAIN'S "ROUGHING IT," HEMINGWAY'S "THE OLD MAN AND THE SEA," FOUR SHORT STORIES, AND 20 LYRIC POEMS. THE SHORT STORIES INCLUDED WERE (1) "THE MONKEY'S PAW" BY W.W. JACOBS, (2) "PAUL'S CASE" BY WILLA CATHER, (3) "THE CASK OF…

75 FR 24786 - List of Approved Spent Fuel Storage Casks: NUHOMS® HD System Revision 1

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-06

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75 FR 33678 - List of Approved Spent Fuel Storage Casks: MAGNASTOR System, Revision 1

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-15

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77 FR 4203 - List of Approved Spent Fuel Storage Casks: MAGNASTOR® System, Revision 2

Federal Register 2010, 2011, 2012, 2013, 2014

2012-01-27

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78 FR 37927 - List of Approved Spent Fuel Storage Casks: MAGNASTOR® System

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-25

...;Prices of new books are listed in the first FEDERAL REGISTER issue of each #0;week. #0; #0; #0; #0;#0... ADAMS Search.'' For problems with ADAMS, please contact the NRC's Public Document Room (PDR) reference... not have a significant economic impact on a substantial number of small entities. This final rule...

76 FR 70331 - List of Approved Spent Fuel Storage Casks: MAGNASTOR ® System, Revision 2

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-14

... various boron-10 areal densities for use with Pressurized Water Reactor and Boiling Water Reactor baskets... add various boron-10 areal densities for use with Pressurized Water Reactor and Boiling Water Reactor....1.1 to add various boron-10 areal densities for use with Pressurized Water Reactor and Boiling Water...

The basic amino acids in the coiled-coil domain of CIN85 regulate its interaction with c-Cbl and phosphatidic acid during epidermal growth factor receptor (EGFR) endocytosis.

PubMed

Zheng, Xiudan; Zhang, Jing; Liao, Kan

2014-07-08

During EGFR internalization CIN85 bridges EGFR-Cbl complex, endocytic machinery and fusible membrane through the interactions of CIN85 with c-Cbl, endophilins and phosphatidic acid. These protein-protein and protein-lipid interactions are mediated or regulated by the positively charged C-terminal coiled-coil domain of CIN85. However, the details of CIN85-lipid interaction remain unknown. The present study suggested a possible electric interaction between the negative charge of phosphatidic acid and the positive charge of basic amino acids in coiled-coil domain. Mutations of the basic amino acids in the coiled-coil domain, especially K645, K646, R648 and R650, into neutral amino acid alanine completely blocked the interaction of CIN85 with c-Cbl or phosphatidic acid. However, they did not affect CIN85-endophilin interaction. In addition, CIN85 was found to associate with the internalized EGFR endosomes. It interacted with several ESCRT (Endosomal Sorting Complex Required for Transport) component proteins for ESCRT assembly on endosomal membrane. Mutations in the coiled-coil domain (deletion of the coiled-coil domain or point mutations of the basic amino acids) dissociated CIN85 from endosomes. These mutants bound the ESCRT components in cytoplasm to prevent them from assembly on endosomal membrane and inhibited EGFR sorting for degradation. As an adaptor protein, CIN85 interacts with variety of partners through several domains. The positive charges of basic amino acids in the coiled-coil domain are not only involved in the interaction with phosphatidic acid, but also regulate the interaction of CIN85 with c-Cbl. CIN85 also interacts with ESCRT components for protein sorting in endosomes. These CIN85-protein and CIN85-lipid interactions enable CIN85 to link EGFR-Cbl endocytic complex with fusible membrane during EGFR endocytosis and subsequently to facilitate ESCRT formation on endosomal membrane for EGFR sorting and degradation.

Denni Algorithm An Enhanced Of SMS (Scan, Move and Sort) Algorithm

NASA Astrophysics Data System (ADS)

Aprilsyah Lubis, Denni; Salim Sitompul, Opim; Marwan; Tulus; Andri Budiman, M.

2017-12-01

Sorting has been a profound area for the algorithmic researchers, and many resources are invested to suggest a more working sorting algorithm. For this purpose many existing sorting algorithms were observed in terms of the efficiency of the algorithmic complexity. Efficient sorting is important to optimize the use of other algorithms that require sorted lists to work correctly. Sorting has been considered as a fundamental problem in the study of algorithms that due to many reasons namely, the necessary to sort information is inherent in many applications, algorithms often use sorting as a key subroutine, in algorithm design there are many essential techniques represented in the body of sorting algorithms, and many engineering issues come to the fore when implementing sorting algorithms., Many algorithms are very well known for sorting the unordered lists, and one of the well-known algorithms that make the process of sorting to be more economical and efficient is SMS (Scan, Move and Sort) algorithm, an enhancement of Quicksort invented Rami Mansi in 2010. This paper presents a new sorting algorithm called Denni-algorithm. The Denni algorithm is considered as an enhancement on the SMS algorithm in average, and worst cases. The Denni algorithm is compared with the SMS algorithm and the results were promising.

Rmax: A systematic approach to evaluate instrument sort performance using center stream catch☆

PubMed Central

Riddell, Andrew; Gardner, Rui; Perez-Gonzalez, Alexis; Lopes, Telma; Martinez, Lola

2015-01-01

Sorting performance can be evaluated with regard to Purity, Yield and/or Recovery of the sorted fraction. Purity is a check on the quality of the sample and the sort decisions made by the instrument. Recovery and Yield definitions vary with some authors regarding both as how efficient the instrument is at sorting the target particles from the original sample, others distinguishing Recovery from Yield, where the former is used to describe the accuracy of the instrument’s sort count. Yield and Recovery are often neglected, mostly due to difficulties in their measurement. Purity of the sort product is often cited alone but is not sufficient to evaluate sorting performance. All of these three performance metrics require re-sampling of the sorted fraction. But, unlike Purity, calculating Yield and/or Recovery calls for the absolute counting of particles in the sorted fraction, which may not be feasible, particularly when dealing with rare populations and precious samples. In addition, the counting process itself involves large errors. Here we describe a new metric for evaluating instrument sort Recovery, defined as the number of particles sorted relative to the number of original particles to be sorted. This calculation requires only measuring the ratios of target and non-target populations in the original pre-sort sample and in the waste stream or center stream catch (CSC), avoiding re-sampling the sorted fraction and absolute counting. We called this new metric Rmax, since it corresponds to the maximum expected Recovery for a particular set of instrument parameters. Rmax is ideal to evaluate and troubleshoot the optimum drop-charge delay of the sorter, or any instrument related failures that will affect sort performance. It can be used as a daily quality control check but can be particularly useful to assess instrument performance before single-cell sorting experiments. Because we do not perturb the sort fraction we can calculate Rmax during the sort process, being especially valuable to check instrument performance during rare population sorts. PMID:25747337

A Novel Mechanism of Regulating the ATPase VPS4 by Its Cofactor LIP5 and the Endosomal Sorting Complex Required for Transport (ESCRT)-III Protein CHMP5

DOE PAGES

Vild, Cody J.; Li, Yan; Guo, Emily Z.; ...

2015-01-30

Disassembly of the endosomal sorting complex required for transport (ESCRT) machinery from biological membranes is a critical final step in cellular processes that require the ESCRT function. This reaction is catalyzed by VPS4, an AAA-ATPase whose activity is tightly regulated by a host of proteins, including LIP5 and the ESCRT-III proteins. In this paper, we present structural and functional analyses of molecular interactions between human VPS4, LIP5, and the ESCRT-III proteins. The N-terminal domain of LIP5 (LIP5NTD) is required for LIP5-mediated stimulation of VPS4, and the ESCRT-III protein CHMP5 strongly inhibits the stimulation. Both of these observations are distinct frommore » what was previously described for homologous yeast proteins. The crystal structure of LIP5NTD in complex with the MIT (microtubule-interacting and transport)-interacting motifs of CHMP5 and a second ESCRT-III protein, CHMP1B, was determined at 1 Å resolution. It reveals an ESCRT-III binding induced moderate conformational change in LIP5NTD, which results from insertion of a conserved CHMP5 tyrosine residue (Tyr 182) at the core of LIP5NTD structure. Finally, mutation of Tyr 182 partially relieves the inhibition displayed by CHMP5. Together, these results suggest a novel mechanism of VPS4 regulation in metazoans, where CHMP5 functions as a negative allosteric switch to control LIP5-mediated stimulation of VPS4.« less

Bone Marrow Regeneration Promoted by Biophysically Sorted Osteoprogenitors From Mesenchymal Stromal Cells

PubMed Central

Poon, Zhiyong; Lee, Wong Cheng; Guan, Guofeng; Nyan, Lin Myint; Lim, Chwee Teck; Han, Jongyoon

2015-01-01

Human tissue repair deficiencies can be supplemented through strategies to isolate, expand in vitro, and reimplant regenerative cells that supplant damaged cells or stimulate endogenous repair mechanisms. Bone marrow-derived mesenchymal stromal cells (MSCs), a subset of which is described as mesenchymal stem cells, are leading candidates for cell-mediated bone repair and wound healing, with hundreds of ongoing clinical trials worldwide. An outstanding key challenge for successful clinical translation of MSCs is the capacity to produce large quantities of cells in vitro with uniform and relevant therapeutic properties. By leveraging biophysical traits of MSC subpopulations and label-free microfluidic cell sorting, we hypothesized and experimentally verified that MSCs of large diameter within expanded MSC cultures were osteoprogenitors that exhibited significantly greater efficacy over other MSC subpopulations in bone marrow repair. Systemic administration of osteoprogenitor MSCs significantly improved survival rates (>80%) as compared with other MSC subpopulations (0%) for preclinical murine bone marrow injury models. Osteoprogenitor MSCs also exerted potent therapeutic effects as “cell factories” that secreted high levels of regenerative factors such as interleukin-6 (IL-6), interleukin-8 (IL-8), vascular endothelial growth factor A, bone morphogenetic protein 2, epidermal growth factor, fibroblast growth factor 1, and angiopoietin-1; this resulted in increased cell proliferation, vessel formation, and reduced apoptosis in bone marrow. This MSC subpopulation mediated rescue of damaged marrow tissue via restoration of the hematopoiesis-supporting stroma, as well as subsequent hematopoiesis. Together, the capabilities described herein for label-freeisolation of regenerative osteoprogenitor MSCs can markedly improve the efficacy of MSC-based therapies. PMID:25411477

The Entangled ER-Mitochondrial axis as a potential therapeutic strategy in Neurodegeneration: A Tangled Duo Unchained

PubMed Central

Joshi, Amit U.; Kornfeld, Opher S.; Mochly-Rosen, Daria

2016-01-01

Endoplasmic reticulum (ER) and mitochondrial function have both been shown to be critical events in neurodegenerative diseases. The ER mediates protein folding, maturation, sorting as well acts as calcium storage. The unfolded protein response (UPR) is a stress response of the ER that is activated by the accumulation of misfolded proteins within the ER lumen. Although the molecular mechanisms underlying ER stress-induced apoptosis are not completely understood, increasing evidence suggests that ER and mitochondria cooperate to signal cell death. Similarly, calcium-mediated mitochondrial function and dynamics not only contribute to ATP generation and calcium buffering but are also a linchpin in mediating cell fate. Mitochondria and ER form structural and functional networks (mitochondria-associated ER membranes [MAMs]) essential to maintaining cellular homeostasis and determining cell fate under various pathophysiological conditions. Regulated Ca2+ transfer from the ER to the mitochondria is important in maintaining control of pro-survival/pro-death pathways. In this review, we summarize the latest therapeutic strategies that target these essential organelles in the context of neurodegenerative diseases. PMID:27212603

Parallel integer sorting with medium and fine-scale parallelism

NASA Technical Reports Server (NTRS)

Dagum, Leonardo

1993-01-01

Two new parallel integer sorting algorithms, queue-sort and barrel-sort, are presented and analyzed in detail. These algorithms do not have optimal parallel complexity, yet they show very good performance in practice. Queue-sort designed for fine-scale parallel architectures which allow the queueing of multiple messages to the same destination. Barrel-sort is designed for medium-scale parallel architectures with a high message passing overhead. The performance results from the implementation of queue-sort on a Connection Machine CM-2 and barrel-sort on a 128 processor iPSC/860 are given. The two implementations are found to be comparable in performance but not as good as a fully vectorized bucket sort on the Cray YMP.

Sort computation

NASA Technical Reports Server (NTRS)

Dorband, John E.

1988-01-01

Sorting has long been used to organize data in preparation for further computation, but sort computation allows some types of computation to be performed during the sort. Sort aggregation and sort distribution are the two basic forms of sort computation. Sort aggregation generates an accumulative or aggregate result for each group of records and places this result in one of the records. An aggregate operation can be any operation that is both associative and commutative, i.e., any operation whose result does not depend on the order of the operands or the order in which the operations are performed. Sort distribution copies the value from a field of a specific record in a group into that field in every record of that group.

ATAC-see reveals the accessible genome by transposase-mediated imaging and sequencing.

PubMed

Chen, Xingqi; Shen, Ying; Draper, Will; Buenrostro, Jason D; Litzenburger, Ulrike; Cho, Seung Woo; Satpathy, Ansuman T; Carter, Ava C; Ghosh, Rajarshi P; East-Seletsky, Alexandra; Doudna, Jennifer A; Greenleaf, William J; Liphardt, Jan T; Chang, Howard Y

2016-12-01

Spatial organization of the genome plays a central role in gene expression, DNA replication, and repair. But current epigenomic approaches largely map DNA regulatory elements outside of the native context of the nucleus. Here we report assay of transposase-accessible chromatin with visualization (ATAC-see), a transposase-mediated imaging technology that employs direct imaging of the accessible genome in situ, cell sorting, and deep sequencing to reveal the identity of the imaged elements. ATAC-see revealed the cell-type-specific spatial organization of the accessible genome and the coordinated process of neutrophil chromatin extrusion, termed NETosis. Integration of ATAC-see with flow cytometry enables automated quantitation and prospective cell isolation as a function of chromatin accessibility, and it reveals a cell-cycle dependence of chromatin accessibility that is especially dynamic in G1 phase. The integration of imaging and epigenomics provides a general and scalable approach for deciphering the spatiotemporal architecture of gene control.

Derivation of sorting programs

NASA Technical Reports Server (NTRS)

Varghese, Joseph; Loganantharaj, Rasiah

1990-01-01

Program synthesis for critical applications has become a viable alternative to program verification. Nested resolution and its extension are used to synthesize a set of sorting programs from their first order logic specifications. A set of sorting programs, such as, naive sort, merge sort, and insertion sort, were successfully synthesized starting from the same set of specifications.

Spin-the-bottle Sort and Annealing Sort: Oblivious Sorting via Round-robin Random Comparisons

PubMed Central

Goodrich, Michael T.

2013-01-01

We study sorting algorithms based on randomized round-robin comparisons. Specifically, we study Spin-the-bottle sort, where comparisons are unrestricted, and Annealing sort, where comparisons are restricted to a distance bounded by a temperature parameter. Both algorithms are simple, randomized, data-oblivious sorting algorithms, which are useful in privacy-preserving computations, but, as we show, Annealing sort is much more efficient. We show that there is an input permutation that causes Spin-the-bottle sort to require Ω(n2 log n) expected time in order to succeed, and that in O(n2 log n) time this algorithm succeeds with high probability for any input. We also show there is a specification of Annealing sort that runs in O(n log n) time and succeeds with very high probability. PMID:24550575

Ultrasonic fingerprinting by phased array transducer

NASA Astrophysics Data System (ADS)

Sednev, D.; Kataeva, O.; Abramets, V.; Pushenko, P.; Tverdokhlebova, T.

2016-06-01

Increasing quantity of spent nuclear fuel that must be under national and international control requires a novel approach to safeguard techniques and equipment. One of the proposed approaches is utilize intrinsic features of casks with spent fuel. In this article an application of a phased array ultrasonic method is considered. This study describes an experimental results on ultrasonic fingerprinting of austenitic steel seam weld.

Instrumentation: Nondestructive Examination for Verification of Canister and Cladding Integrity. FY2014 Status Update

DOE Office of Scientific and Technical Information (OSTI.GOV)

Meyer, Ryan M.; Suter, Jonathan D.; Jones, Anthony M.

2014-09-12

This report documents FY14 efforts for two instrumentation subtasks under storage and transportation. These instrumentation tasks relate to developing effective nondestructive evaluation (NDE) methods and techniques to (1) verify the integrity of metal canisters for the storage of used nuclear fuel (UNF) and to (2) verify the integrity of dry storage cask internals.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ingham, J.G.

The IDENT 1578 container, which is a 110-in. long 5.5-in. OD tube, is designed for shipping FFTF fuel elements in T-3 casks between HEDL, HFEF, and other laboratories. The thermal analysis was conducted to evaluate whether or not the container satisfies its thermal design criteria (handle a decay heat load of 600 watts, max fuel pin cladding temperature not exceeding 800/sup 0/F).

ROUGHING IT. THE OLD MAN AND THE SEA. SHORT STORIES. LYRIC POETRY. LITERATURE CURRICULUM III, STUDENT VERSION.

ERIC Educational Resources Information Center

KITZHABER, ALBERT R.

A STUDENT VERSION OF A LITERATURE CURRICULUM GUIDE WAS PROVIDED FOR TWAIN'S "ROUGHING IT," HEMINGWAY'S "THE OLD MAN AND THE SEA," FOUR SHORT STORIES, AND 20 LYRIC POEMS. THE SHORT STORIES INCLUDED WERE (1) "THE MONKEY'S PAW" BY W.W. JACOBS, (2) "PAUL'S CASE" BY WILLA CATHER, (3) "THE CASK OF…

75 FR 23820 - Notice of Docketing of Amendment Request for Materials License No. SNM-2506; Northern States...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-04

... INFORMATION CONTACT: Pamela Longmire, Ph.D., Project Manager, Licensing Branch, Division of Spent Fuel Storage... Generating Plant (PINGP), Unit Nos. 1 and 2, site in Goodhue County, Minnesota. The TN-40 cask is currently..., higher burnup spent fuel used in the PINGP reactor as well as associated changes to the ISFSI's technical...

Constructing Complexity: Using Reading Levels to Differentiate Reading Comprehension Activities

ERIC Educational Resources Information Center

FitzPatrick, Declan

2008-01-01

The author remembers a class when he asked his students to discuss in small groups how Edgar Allan Poe suggests a judgment of the main character in "The Cask of Amontillado". During their discussion it became clear to the author that the students couldn't come to consensus because they had no grasp of the narrator's explanations of his motivations…

75 FR 33853 - Maine Yankee Atomic Power Company; Independent Spent Fuel Storage Installation; Issuance of...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-15

...) Surveillance Requirement 3.1.6.1 to verify the operability of the concrete cask heat removal system to maintain... Amendment No. 5 for one storage canister at the MY ISFSI. The affected storage canister had a heat load of 9..., and the LCO 3.1.4 time limit for a canister [[Page 33855

Recent developments - US spent fuel disposition

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

One of a US utility's major risk factors in continuing to operate a nuclear plant is managing discharged spent fuel. The US Department of Energy (DOE) signed contracts with utilities guaranteeing government acceptance of spent fuel by 1988. However, on December 17, 1992, DOE Secretary Watkins wrote to Sen. J. Bennett Johnston (D-LA), Chairman of the Senate Energy Committee, indicating a reassessment of DOE's programs, the results of which will be presented to Congress in January 1993. He indicated the Department may not be able to meet the 1988 date, because of difficulty in finding a site for the Monitoredmore » Retrievable Storage facility. Watkins indicated that DOE has investigated an interim solution and decided to expedite a program to certify a multi-purpose standardized cask system for spent fuel receipt, storage, transport, and disposal. To meet the expectations of US utilities, DOE is considering a plan to use federal sites for interim storage of the casks. Secretary Watkins recommended the waste program be taken off-budget and put in a revolving fund established to ensure that money already collected from utilities will be available to meet the schedule for completion of the repository.« less

SRNL Development of Recovery Processes for Mark-18A Heavy Actinide Targets

DOE Office of Scientific and Technical Information (OSTI.GOV)

Allender, Jeffrey S.; Bridges, Nicholas J.; Loftin, Bradley M.

2015-07-14

Savannah River National Laboratory (SRNL) and Oak Ridge National Laboratory (ORNL) are developing plans for the recovery of rare and unique isotopes contained within heavy-actinide target assemblies, specifically the Mark-18A. Mark-18A assemblies were irradiated in Savannah River Site (SRS) reactors in the 1970s under extremely high neutron-flux conditions and produced, virtually, the world's supply of plutonium-244, an isotope of key importance to high-precision actinide measurement and other scientific and nonproliferation uses; and curium highly enriched in heavy isotopes (e.g., curium-246 and curium-248). In 2015 and 2016, SRNL is pursuing tasks that would reduce program risk and budget requirements, including furthermore » characterization of unprocessed targets; engineering studies for the use of the SRNL Shielded Cells Facility (SCF) for recovery; and development of onsite and offsite shipping methods including a replacement for the heavy (70 ton) cask previously used for onsite transfer of irradiated items at SRS. A status update is provided for the characterization, including modeling using the Monte Carlo N-Particle Transport Code (MCNP); direct non-destructive assay measurements; and cask design.« less

Development of a novel ultrasonic temperature probe for long-term monitoring of dry cask storage systems

NASA Astrophysics Data System (ADS)

Bakhtiari, S.; Wang, K.; Elmer, T. W.; Koehl, E.; Raptis, A. C.

2013-01-01

With the recent cancellation of the Yucca Mountain repository and the limited availability of wet storage utilities for spent nuclear fuel (SNF), more attention has been directed toward dry cask storage systems (DCSSs) for long-term storage of SNF. Consequently, more stringent guidelines have been issued for the aging management of dry storage facilities that necessitate monitoring of the conditions of DCSSs. Continuous health monitoring of DCSSs based on temperature variations is one viable method for assessing the integrity of the system. In the present work, a novel ultrasonic temperature probe (UTP) is being tested for long-term online temperature monitoring of DCSSs. Its performance was evaluated and compared with type N thermocouple (NTC) and resistance temperature detector (RTD) using a small-scale dry storage canister mockup. Our preliminary results demonstrate that the UTP system developed at Argonne is able to achieve better than 0.8 °C accuracy, tested at temperatures of up to 400 °C. The temperature resolution is limited only by the sampling rate of the current system. The flexibility of the probe allows conforming to complex geometries thus making the sensor particularly suited to measurement scenarios where access is limited.

Electromagnetic Acoustic Transducers for Robotic Nondestructive Inspection in Harsh Environments.

PubMed

Choi, Sungho; Cho, Hwanjeong; Lindsey, Matthew S; Lissenden, Cliff J

2018-01-11

Elevated temperature, gamma radiation, and geometric constraints inside dry storage casks for spent nuclear fuel represent a harsh environment for nondestructive inspection of the cask and require that the inspection be conducted with a robotic system. Electromagnetic acoustic transducers (EMATs) using non-contact ultrasonic transduction based on the Lorentz force to excite/receive ultrasonic waves are suited for use in the robotic inspection. Periodic permanent magnet EMATs that actuate/receive shear horizontal guided waves are developed for application to robotic nondestructive inspection of stress corrosion cracks in the heat affected zone of welds in stainless steel dry storage canisters. The EMAT's components are carefully selected in consideration of the inspection environment, and tested under elevated temperature and gamma radiation doses up to 177 °C and 5920 krad, respectively, to evaluate the performance of the EMATs under realistic environmental conditions. The effect of gamma radiation is minimal, but the EMAT's performance is affected by temperatures above 121 °C due to the low Curie temperature of the magnets. Different magnets are needed to operate at 177 °C. The EMAT's capability to detect notches is also evaluated from B-scan measurements on 304 stainless steel welded plate containing surface-breaking notches.

Safe sorting of GFP-transduced live cells for subsequent culture using a modified FACS vantage.

PubMed

Sørensen, T U; Gram, G J; Nielsen, S D; Hansen, J E

1999-12-01

A stream-in-air cell sorter enables rapid sorting to a high purity, but it is not well suited for sorting of infectious material due to the risk of airborne spread to the surroundings. A FACS Vantage cell sorter was modified for safe use with potentially HIV infected cells. Safety tests with bacteriophages were performed to evaluate the potential spread of biologically active material during cell sorting. Cells transduced with a retroviral vector carrying the gene for GFP were sorted on the basis of their GFP fluorescence, and GFP expression was followed during subsequent culture. The bacteriophage sorting showed that the biologically active material was confined to the sorting chamber. A failure mode simulating a nozzle blockage resulted in detectable droplets inside the sorting chamber, but no droplets could be detected when an additional air suction from the sorting chamber had been put on. The GFP transduced cells were sorted to 99% purity. Cells not expressing GFP at the time of sorting did not turn on the gene during subsequent culture. Un-sorted cells and cells sorted to be positive for GFP showed a decrease in the fraction of GFP positive cells during culture. Sorting of live infected cells can be performed safely and with no deleterious effects on vector expression using the modified FACS Vantage instrument. Copyright 1999 Wiley-Liss, Inc.

SNX10 mediates alcohol-induced liver injury and steatosis by regulating the activation of chaperone-mediated autophagy.

PubMed

You, Yan; Li, Wan-Zhen; Zhang, Sulin; Hu, Bin; Li, Yue-Xuan; Li, Hai-Dong; Tang, Huan-Huan; Li, Qian-Wen; Guan, Yun-Yun; Liu, Li-Xin; Bao, Wei-Lian; Shen, Xiaoyan

2018-07-01

Alcoholic liver disease (ALD) is a major cause of morbidity and mortality worldwide. However, the cellular defense mechanisms underlying ALD are not well understood. Recent studies highlighted the involvement of chaperone-mediated autophagy (CMA) in regulating hepatic lipid metabolism. Sorting nexin (SNX)-10 has a regulatory function in endolysosomal trafficking and stabilisation. Here, we investigated the roles of SNX10 in CMA activation and in the pathogenesis of alcohol-induced liver injury and steatosis. Snx10 knockout (Snx10 KO) mice and their wild-type (WT) littermates fed either the Lieber-DeCarli liquid alcohol diet or a control liquid diet, and primary cultured WT and Snx10 KO hepatocytes stimulated with ethanol, were used as in vivo and in vitro ALD models, respectively. Activation of CMA, liver injury parameters, inflammatory cytokines, oxidative stress and lipid metabolism were measured. Compared with WT littermates, Snx10 KO mice exhibited a significant amelioration in ethanol-induced liver injury and hepatic steatosis. Both in vivo and in vitro studies showed that SNX10 deficiency upregulated lysosome-associated membrane protein type 2A (LAMP-2A) expression and CMA activation, which could be reversed by SNX10 overexpression in vitro. LAMP-2A interference confirmed that the upregulation of Nrf2 and AMPK signalling pathways induced by SNX10 deficiency relied on CMA activation. Pull-down assays revealed an interaction between SNX10 and cathepsin A (CTSA), a key enzyme involved in LAMP-2A degradation. Deficiency in SNX10 inhibited CTSA maturation and increased the stability of LAMP-2A, resulting in an increase in CMA activity. SNX10 controls CMA activity by mediating CTSA maturation, and, thus, has an essential role in alcohol-induced liver injury and steatosis. Our results provide evidence for SNX10 as a potential promising therapeutic target for preventing or ameliorating liver injury in ALD. Alcoholic liver disease is a major cause of morbidity and mortality worldwide. Recent studies highlight the involvement of chaperone-mediated autophagy (CMA) in regulating hepatic lipid metabolism. Our study reveals that deficiency of sorting nexin (SNX) 10 increases the stability of LAMP-2A by inhibiting cathepsin A maturation, resulting in the increase of CMA activity and, thus, alleviates alcohol-induced liver injury and steatosis. Copyright © 2018 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Design and realization of sort manipulator of crystal-angle sort machine

NASA Astrophysics Data System (ADS)

Wang, Ming-shun; Chen, Shu-ping; Guan, Shou-ping; Zhang, Yao-wei

2005-12-01

It is a current tendency of development in automation technology to replace manpower with manipulators in working places where dangerous, harmful, heavy or repetitive work is involved. The sort manipulator is installed in a crystal-angle sort machine to take the place of manpower, and engaged in unloading and sorting work. It is the outcome of combing together mechanism, electric transmission, and pneumatic element and micro-controller control. The step motor makes the sort manipulator operate precisely. The pneumatic elements make the sort manipulator be cleverer. Micro-controller's software bestows some simple artificial intelligence on the sort manipulator, so that it can precisely repeat its unloading and sorting work. The combination of manipulator's zero position and step motor counting control puts an end to accumulating error in long time operation. A sort manipulator's design in the practice engineering has been proved to be correct and reliable.

PDZ Ligand Binding-Induced Conformational Coupling of the PDZ-SH3-GK Tandems in PSD-95 Family MAGUKs.

PubMed

Zeng, Menglong; Ye, Fei; Xu, Jia; Zhang, Mingjie

2018-01-05

Discs large (DLG) MAGUKs are abundantly expressed in glutamatergic synapses, crucial for synaptic transmission, and plasticity by anchoring various postsynaptic components including glutamate receptors, downstream scaffold proteins and signaling enzymes. Different DLG members have shared structures and functions, but also contain unique features. How DLG family proteins function individually and cooperatively is largely unknown. Here, we report that PSD-95 PDZ3 directly couples with SH3-GK tandem in a PDZ ligand binding-dependent manner, and the coupling can promote PSD-95 dimerization and multimerization. Aided by sortase-mediated protein ligation and selectively labeling, we elucidated the PDZ3/SH3-GK conformational coupling mechanism using NMR spectroscopy. We further demonstrated that PSD-93, but not SAP102, can also undergo PDZ3 ligand binding-induced conformational coupling with SH3-GK and form homo-oligomers. Interestingly, PSD-95 and PSD-93 can also form ligand binding-induced hetero-oligomers, suggesting a cooperative assembly mechanism for the mega-N-methyl-d-aspartate receptor synaptic signaling complex. Finally, we provide evidence showing that ligand binding-induced conformational coupling between PDZ and SH3-GK is a common feature for other MAGUKs including CASK and PALS1. Copyright © 2017 Elsevier Ltd. All rights reserved.

In vitro selection of zinc fingers with altered DNA-binding specificity.

PubMed

Jamieson, A C; Kim, S H; Wells, J A

1994-05-17

We have used random mutagenesis and phage display to alter the DNA-binding specificity of Zif268, a transcription factor that contains three zinc finger domains. Four residues in the helix of finger 1 of Zif268 that potentially mediate DNA binding were identified from an X-ray structure of the Zif268-DNA complex. A library was constructed in which these residues were randomly mutated and the Zif268 variants were fused to a truncated version of the gene III coat protein on the surface of M13 filamentous phage particles. The phage displayed the mutant proteins in a monovalent fashion and were sorted by repeated binding and elution from affinity matrices containing different DNA sequences. When the matrix contained the natural nine base pair operator sequence 5'-GCG-TGG-GCG-3', native-like zinc fingers were isolated. New finger 1 variants were found by sorting with two different operators in which the singly modified triplets, GTG and TCG, replaced the native finger 1 triplet, GCG. Overall, the selected finger 1 variants contained a preponderance of polar residues at the four sites. Interestingly, the net charge of the four residues in any selected finger never derived more that one unit from neutrality despite the fact that about half the variants contained three or four charged residues over the four sites. Measurements of the dissociation constants for two of these purified finger 1 variants by gel-shift assay showed their specificities to vary over a 10-fold range, with the greatest affinity being for the DNA binding site for which they were sorted.(ABSTRACT TRUNCATED AT 250 WORDS)

Elastomeric microparticles for acoustic mediated bioseparations

PubMed Central

2013-01-01

Background Acoustophoresis has been utilized successfully in applications including cell trapping, focusing, and purification. One current limitation of acoustophoresis for cell sorting is the reliance on the inherent physical properties of cells (e.g., compressibility, density) instead of selecting cells based upon biologically relevant surface-presenting antigens. Introducing an acoustophoretic cell sorting approach that allows biochemical specificity may overcome this limitation, thus advancing the value of acoustophoresis approaches for both the basic research and clinical fields. Results The results presented herein demonstrate the ability for negative acoustic contrast particles (NACPs) to specifically capture and transport positive acoustic contrast particles (PACPs) to the antinode of an ultrasound standing wave. Emulsification and post curing of pre-polymers, either polydimethylsiloxane (PDMS) or polyvinylmethylsiloxane (PVMS), within aqueous surfactant solution results in the formation of stable NACPs that focus onto pressure antinodes. We used either photochemical reactions with biotin-tetrafluorophenyl azide (biotin-TFPA) or end-functionalization of Pluronic F108 surfactant to biofunctionalize NACPs. These biotinylated NACPs bind specifically to streptavidin polystyrene microparticles (as cell surrogates) and transport them to the pressure antinode within an acoustofluidic chip. Conclusion To the best of our knowledge, this is the first demonstration of using NACPs as carriers for transport of PACPs in an ultrasound standing wave. By using different silicones (i.e., PDMS, PVMS) and curing chemistries, we demonstrate versatility of silicone materials for NACPs and advance the understanding of useful approaches for preparing NACPs. This bioseparation scheme holds potential for applications requiring rapid, continuous separations such as sorting and analysis of cells and biomolecules. PMID:23809852

DLK1 as a potential target against cancer stem/progenitor cells of hepatocellular carcinoma.

PubMed

Xu, Xiao; Liu, Rui-Fang; Zhang, Xin; Huang, Li-Yu; Chen, Fei; Fei, Qian-Lan; Han, Ze-Guang

2012-03-01

Delta-like 1 homolog (DLK1; Drosophila) is a hepatic stem/progenitor cell marker in fetal livers that plays a vital role in oncogenesis of hepatocellular carcinoma (HCC). The aim of this study is to investigate whether DLK1 could serve as a potential therapeutic target against cancer stem/progenitor cells of HCC. DLK1(+) and DLK1(-) cells were sorted by fluorescence-activated cell sorting and magnetic-activated cell sorting, respectively, and then were evaluated by flow cytometry. The biological behaviors of these isolated cells and those with DLK1 knockdown were assessed by growth curve, colony formation assay, spheroid colony formation, chemoresistance, and in vivo tumorigenicity. Adenovirus-mediated RNA interference was used to knockdown the endogenous DLK1. We found that DLK1(+) population was less than 10% in almost all 17 HCC cell lines examined. DLK1(+) HCC cells showed stronger ability of chemoresistance, colony formation, spheroid colony formation, and in vivo tumorigenicity compared with DLK1(-) cells. The DLK1(+) HCC cells could generate the progeny without DLK1 expression. Furthermore, DLK1 knockdown could suppress the ability of proliferation, colony formation, spheroid colony formation, and in vivo tumorigenicity of Hep3B and Huh-7 HCC cells. Our data suggested that DLK1(+) HCC cells have characteristics similar to those of cancer stem/progenitor cells. RNA interference against DLK1 can suppress the malignant behaviors of HCC cells, possibly through directly disrupting cancer stem/progenitor cells, which suggested that DLK1 could be a potential therapeutic target against the HCC stem/progenitor cells.

A Task Analysis of the Shift from Teacher Instructions to Self-Instructions in Performing an In-Common Task.

ERIC Educational Resources Information Center

Grote, Irene; And Others

1996-01-01

Three preschoolers performed four sorts with stimulus cards--an untaught target sort and three directly taught alternating sorts considered to self-instruct the target performance. Accuracy increased first in the skill sorts and then in the untaught target sorts. All subjects generalized to new target sorts. Correct spontaneous self-instructions…

To sort or not to sort: the impact of spike-sorting on neural decoding performance.

PubMed

Todorova, Sonia; Sadtler, Patrick; Batista, Aaron; Chase, Steven; Ventura, Valérie

2014-10-01

Brain-computer interfaces (BCIs) are a promising technology for restoring motor ability to paralyzed patients. Spiking-based BCIs have successfully been used in clinical trials to control multi-degree-of-freedom robotic devices. Current implementations of these devices require a lengthy spike-sorting step, which is an obstacle to moving this technology from the lab to the clinic. A viable alternative is to avoid spike-sorting, treating all threshold crossings of the voltage waveform on an electrode as coming from one putative neuron. It is not known, however, how much decoding information might be lost by ignoring spike identity. We present a full analysis of the effects of spike-sorting schemes on decoding performance. Specifically, we compare how well two common decoders, the optimal linear estimator and the Kalman filter, reconstruct the arm movements of non-human primates performing reaching tasks, when receiving input from various sorting schemes. The schemes we tested included: using threshold crossings without spike-sorting; expert-sorting discarding the noise; expert-sorting, including the noise as if it were another neuron; and automatic spike-sorting using waveform features. We also decoded from a joint statistical model for the waveforms and tuning curves, which does not involve an explicit spike-sorting step. Discarding the threshold crossings that cannot be assigned to neurons degrades decoding: no spikes should be discarded. Decoding based on spike-sorted units outperforms decoding based on electrodes voltage crossings: spike-sorting is useful. The four waveform based spike-sorting methods tested here yield similar decoding efficiencies: a fast and simple method is competitive. Decoding using the joint waveform and tuning model shows promise but is not consistently superior. Our results indicate that simple automated spike-sorting performs as well as the more computationally or manually intensive methods used here. Even basic spike-sorting adds value to the low-threshold waveform-crossing methods often employed in BCI decoding.

To sort or not to sort: the impact of spike-sorting on neural decoding performance

NASA Astrophysics Data System (ADS)

Todorova, Sonia; Sadtler, Patrick; Batista, Aaron; Chase, Steven; Ventura, Valérie

2014-10-01

Objective. Brain-computer interfaces (BCIs) are a promising technology for restoring motor ability to paralyzed patients. Spiking-based BCIs have successfully been used in clinical trials to control multi-degree-of-freedom robotic devices. Current implementations of these devices require a lengthy spike-sorting step, which is an obstacle to moving this technology from the lab to the clinic. A viable alternative is to avoid spike-sorting, treating all threshold crossings of the voltage waveform on an electrode as coming from one putative neuron. It is not known, however, how much decoding information might be lost by ignoring spike identity. Approach. We present a full analysis of the effects of spike-sorting schemes on decoding performance. Specifically, we compare how well two common decoders, the optimal linear estimator and the Kalman filter, reconstruct the arm movements of non-human primates performing reaching tasks, when receiving input from various sorting schemes. The schemes we tested included: using threshold crossings without spike-sorting; expert-sorting discarding the noise; expert-sorting, including the noise as if it were another neuron; and automatic spike-sorting using waveform features. We also decoded from a joint statistical model for the waveforms and tuning curves, which does not involve an explicit spike-sorting step. Main results. Discarding the threshold crossings that cannot be assigned to neurons degrades decoding: no spikes should be discarded. Decoding based on spike-sorted units outperforms decoding based on electrodes voltage crossings: spike-sorting is useful. The four waveform based spike-sorting methods tested here yield similar decoding efficiencies: a fast and simple method is competitive. Decoding using the joint waveform and tuning model shows promise but is not consistently superior. Significance. Our results indicate that simple automated spike-sorting performs as well as the more computationally or manually intensive methods used here. Even basic spike-sorting adds value to the low-threshold waveform-crossing methods often employed in BCI decoding.

Antigen-mediated regulation in monoclonal gammopathies and myeloma

PubMed Central

Nair, Shiny; Sng, Joel; Boddupalli, Chandra Sekhar; Seckinger, Anja; Fulciniti, Mariateresa; Zhang, Lin; Rauniyar, Navin; Lopez, Michael; Neparidze, Natalia; Parker, Terri; Munshi, Nikhil C.; Sexton, Rachael; Barlogie, Bart; Orlowski, Robert; Bergsagel, Leif; Hose, Dirk; Mistry, Pramod K.; Meffre, Eric; Dhodapkar, Madhav V.

2018-01-01

A role for antigen-driven stimulation has been proposed in the pathogenesis of monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM) based largely on the binding properties of monoclonal Ig. However, insights into antigen binding to clonal B cell receptors and in vivo responsiveness of the malignant clone to antigen-mediated stimulation are needed to understand the role of antigenic stimulation in tumor growth. Lysolipid-reactive clonal Ig were detected in Gaucher disease (GD) and some sporadic gammopathies. Here, we show that recombinant Ig (rIg) cloned from sort-purified single tumor cells from lipid-reactive sporadic and GD-associated gammopathy specifically bound lysolipids. Liposome sedimentation and binding assays confirmed specific interaction of lipid-reactive monoclonal Ig with lysolipids. The clonal nature of lysolipid-binding Ig was validated by protein sequencing. Gene expression profiling and cytogenetic analyses from 2 patient cohorts showed enrichment of nonhyperdiploid tumors in lipid-reactive patients. In vivo antigen-mediated stimulation led to an increase in clonal Ig and plasma cells (PCs) in GD gammopathy and also reactivated previously suppressed antigenically related nonclonal PCs. These data support a model wherein antigenic stimulation mediates an initial polyclonal phase, followed by evolution of monoclonal tumors enriched in nonhyperdiploid genomes, responsive to underlying antigen. Targeting underlying antigens may therefore prevent clinical MM. PMID:29669929

Antigen-mediated regulation in monoclonal gammopathies and myeloma.

PubMed

Nair, Shiny; Sng, Joel; Boddupalli, Chandra Sekhar; Seckinger, Anja; Chesi, Marta; Fulciniti, Mariateresa; Zhang, Lin; Rauniyar, Navin; Lopez, Michael; Neparidze, Natalia; Parker, Terri; Munshi, Nikhil C; Sexton, Rachael; Barlogie, Bart; Orlowski, Robert; Bergsagel, Leif; Hose, Dirk; Flavell, Richard A; Mistry, Pramod K; Meffre, Eric; Dhodapkar, Madhav V

2018-04-19

A role for antigen-driven stimulation has been proposed in the pathogenesis of monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM) based largely on the binding properties of monoclonal Ig. However, insights into antigen binding to clonal B cell receptors and in vivo responsiveness of the malignant clone to antigen-mediated stimulation are needed to understand the role of antigenic stimulation in tumor growth. Lysolipid-reactive clonal Ig were detected in Gaucher disease (GD) and some sporadic gammopathies. Here, we show that recombinant Ig (rIg) cloned from sort-purified single tumor cells from lipid-reactive sporadic and GD-associated gammopathy specifically bound lysolipids. Liposome sedimentation and binding assays confirmed specific interaction of lipid-reactive monoclonal Ig with lysolipids. The clonal nature of lysolipid-binding Ig was validated by protein sequencing. Gene expression profiling and cytogenetic analyses from 2 patient cohorts showed enrichment of nonhyperdiploid tumors in lipid-reactive patients. In vivo antigen-mediated stimulation led to an increase in clonal Ig and plasma cells (PCs) in GD gammopathy and also reactivated previously suppressed antigenically related nonclonal PCs. These data support a model wherein antigenic stimulation mediates an initial polyclonal phase, followed by evolution of monoclonal tumors enriched in nonhyperdiploid genomes, responsive to underlying antigen. Targeting underlying antigens may therefore prevent clinical MM.

[Food allergy, food intolerance or functional disorder?].

PubMed

Wüthrich, B

2009-04-01

The term "food allergy" is widely misused for all sorts of symptoms and diseases caused by food. Food allergy (FA) is an adverse reaction to food (food hypersensitivity) occurring in susceptible individuals, which is mediated by a classical immune mechanism specific for the food itself. The best established mechanism in FA is due to the presence of IgE antibodies against the offending food. Food intolerance (FI) are all non-immune-mediated adverse reactions to food. The subgroups of FI are enzymatic (e.g. lactose intolerance due to lactase deficiency), pharmacological (reactions against biogenic amines, histamine intolerance), and undefined food intolerance (e.g. against some food additives). The diagnosis of an IgE-mediated FA is made by a carefully taken case history, supported by the demonstration of an IgE sensitization either by skin prick tests or by in vitro tests, and confirmed by positive oral provocation. For scientific purposes the only accepted test for the confirmation of FA/FI is a properly performed double-blind, placebo-controlled food challenge (DBPCFC). A panel of recombinant allergens, produced as single allergenic molecules, may in future improve the diagnosis of IgE-mediated FA. Due to a lack of causal treatment possibilities, the elimination of the culprit "food allergen" from the diet is the only therapeutic option for patients with real food allergy.

Design of monitoring system for mail-sorting based on the Profibus S7 series PLC

NASA Astrophysics Data System (ADS)

Zhang, W.; Jia, S. H.; Wang, Y. H.; Liu, H.; Tang, G. C.

2017-01-01

With the rapid development of the postal express, the workload of mail sorting is increasing, but the automatic technology of mail sorting is not mature enough. In view of this, the system uses Siemens S7-300 PLC as the main station controller, PLC of Siemens S7-200/400 is from the station controller, through the man-machine interface configuration software MCGS, PROFIBUS-DP communication, RFID technology and mechanical sorting hand achieve mail classification sorting monitoring. Among them, distinguish mail-sorting by scanning RFID posted in the mail electronic bar code (fixed code), the system uses the corresponding controller on the acquisition of information processing, the processed information transmit to the sorting manipulator by PROFIBUS-DP. The system can realize accurate and efficient mail sorting, which will promote the development of mail sorting technology.

Dissecting Stop Transfer versus Conservative Sorting Pathways for Mitochondrial Inner Membrane Proteins in Vivo*

PubMed Central

Park, Kwangjin; Botelho, Salomé Calado; Hong, Joonki; Österberg, Marie; Kim, Hyun

2013-01-01

Mitochondrial inner membrane proteins that carry an N-terminal presequence are sorted by one of two pathways: stop transfer or conservative sorting. However, the sorting pathway is known for only a small number of proteins, in part due to the lack of robust experimental tools with which to study. Here we present an approach that facilitates determination of inner membrane protein sorting pathways in vivo by fusing a mitochondrial inner membrane protein to the C-terminal part of Mgm1p containing the rhomboid cleavage region. We validated the Mgm1 fusion approach using a set of proteins for which the sorting pathway is known, and determined sorting pathways of inner membrane proteins for which the sorting mode was previously uncharacterized. For Sdh4p, a multispanning membrane protein, our results suggest that both conservative sorting and stop transfer mechanisms are required for insertion. Furthermore, the sorting process of Mgm1 fusion proteins was analyzed under different growth conditions and yeast mutant strains that were defective in the import motor or the m-AAA protease function. Our results show that the sorting of mitochondrial proteins carrying moderately hydrophobic transmembrane segments is sensitive to cellular conditions, implying that mitochondrial import and membrane sorting in the physiological environment may be dynamically tuned. PMID:23184936

Seminal plasma affects sperm sex sorting in boars.

PubMed

Alkmin, Diego V; Parrilla, Inmaculada; Tarantini, Tatiana; Del Olmo, David; Vazquez, Juan M; Martinez, Emilio A; Roca, Jordi

2016-04-01

Two experiments were conducted in boar semen samples to evaluate how both holding time (24h) and the presence of seminal plasma (SP) before sorting affect sperm sortability and the ability of sex-sorted spermatozoa to tolerate liquid storage. Whole ejaculate samples were divided into three aliquots immediately after collection: one was diluted (1:1, v/v) in Beltsville thawing solution (BTS; 50% SP); the SP of the other two aliquots was removed and the sperm pellets were diluted with BTS + 10% of their own SP (10% SP) or BTS alone (0% SP). The three aliquots of each ejaculate were divided into two portions, one that was processed immediately for sorting and a second that was sorted after 24h storage at 15-17°C. In the first experiment, the ability to exhibit well-defined X- and Y-chromosome-bearing sperm peaks (split) in the cytometry histogram and the subsequent sorting efficiency were assessed (20 ejaculates). In contrast with holding time, the SP proportion influenced the parameters examined, as evidenced by the higher number of ejaculates exhibiting split and better sorting efficiency (P<0.05) in semen samples with 0-10% SP compared with those with 50% SP. In a second experiment, the quality (viability, total and progressive motility) and functionality (plasma membrane fluidity and intracellular generation of reactive oxygen species) of sex-sorted spermatozoa were evaluated after 0, 72 and 120h storage at 15-17°C (10 ejaculates). Holding time and SP proportion did not influence the quality or functionality of stored sex-sorted spermatozoa. In conclusion, a holding time as long as 24h before sorting did not negatively affect sex sorting efficiency or the ability of sorted boar spermatozoa to tolerate long-term liquid storage. A high proportion of SP (50%) in the semen samples before sorting reduced the number of ejaculates to be sorted and negatively influenced the sorting efficiency, but did not affect the ability of sex-sorted spermatozoa to tolerate liquid storage.