Sample records for clamp loader complex

  1. Mechanism of opening a sliding clamp

    PubMed Central

    Douma, Lauren G.; Yu, Kevin K.; England, Jennifer K.

    2017-01-01

    Abstract Clamp loaders load ring-shaped sliding clamps onto DNA where the clamps serve as processivity factors for DNA polymerases. In the first stage of clamp loading, clamp loaders bind and stabilize clamps in an open conformation, and in the second stage, clamp loaders place the open clamps around DNA so that the clamps encircle DNA. Here, the mechanism of the initial clamp opening stage is investigated. Mutations were introduced into the Escherichia coli β-sliding clamp that destabilize the dimer interface to determine whether the formation of an open clamp loader–clamp complex is dependent on spontaneous clamp opening events. In other work, we showed that mutation of a positively charged Arg residue at the β-dimer interface and high NaCl concentrations destabilize the clamp, but neither facilitates the formation of an open clamp loader–clamp complex in experiments presented here. Clamp opening reactions could be fit to a minimal three-step ‘bind-open-lock’ model in which the clamp loader binds a closed clamp, the clamp opens, and subsequent conformational rearrangements ‘lock’ the clamp loader–clamp complex in a stable open conformation. Our results support a model in which the E. coli clamp loader actively opens the β-sliding clamp. PMID:28973453

  2. Structural analysis of a eukaryotic sliding DNA clamp-clamp loadercomplex.

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

    Bowman, Gregory D.; O'Donnell, Mike; Kuriyan, John

    2006-06-17

    Sliding clamps are ring-shaped proteins that encircle DNA and confer high processivity on DNA polymerases. Here we report the crystal structure of the five-protein clamp loader complex (replication factor-C, RFC) of the yeast Saccharomyces cerevisiae, bound to the sliding clamp (proliferating cell nuclear antigen, PCNA). Tight interfacial coordination of the ATP analogue ATP-?-S by RFC results in a spiral arrangement of the ATPase domains of the clamp loader above the PCNA ring. Placement of a model for primed DNA within the central hole of PCNA reveals a striking correspondence between the RFC spiral and the grooves of the DNA doublemore » helix. This model, in which the clamp loader complex locks onto primed DNA in a screw-cap-like arrangement, provides a simple explanation for the process by which the engagement of primer-template junctions by the RFC:PCNA complex results in ATP hydrolysis and release of the sliding clamp on DNA.« less

  3. Insights into the structure and assembly of the Bacillus subtilis clamp-loader complex and its interaction with the replicative helicase

    PubMed Central

    Afonso, José P.; Chintakayala, Kiran; Suwannachart, Chatrudee; Sedelnikova, Svetlana; Giles, Kevin; Hoyes, John B.; Soultanas, Panos; Rafferty, John B.; Oldham, Neil J.

    2013-01-01

    The clamp-loader complex plays a crucial role in DNA replication by loading the β-clamp onto primed DNA to be used by the replicative polymerase. Relatively little is known about the stoichiometry, structure and assembly pathway of this complex, and how it interacts with the replicative helicase, in Gram-positive organisms. Analysis of full and partial complexes by mass spectrometry revealed that a hetero-pentameric τ3-δ-δ′ Bacillus subtilis clamp-loader assembles via multiple pathways, which differ from those exhibited by the Gram-negative model Escherichia coli. Based on this information, a homology model of the B. subtilis τ3-δ-δ′ complex was constructed, which revealed the spatial positioning of the full C-terminal τ domain. The structure of the δ subunit was determined by X-ray crystallography and shown to differ from that of E. coli in the nature of the amino acids comprising the τ and δ′ binding regions. Most notably, the τ-δ interaction appears to be hydrophilic in nature compared with the hydrophobic interaction in E. coli. Finally, the interaction between τ3 and the replicative helicase DnaB was driven by ATP/Mg2+ conformational changes in DnaB, and evidence is provided that hydrolysis of one ATP molecule by the DnaB hexamer is sufficient to stabilize its interaction with τ3. PMID:23525462

  4. The structure of a ring-opened proliferating cell nuclear antigen-replication factor C complex revealed by fluorescence energy transfer.

    PubMed

    Zhuang, Zhihao; Yoder, Bonita L; Burgers, Peter M J; Benkovic, Stephen J

    2006-02-21

    Numerous proteins that function in DNA metabolic pathways are known to interact with the proliferating cell nuclear antigen (PCNA). The important function of PCNA in stimulating various cellular activities requires its topological linkage with DNA. Loading of the circular PCNA onto duplex DNA requires the activity of a clamp-loader [replication factor C (RFC)] complex and the energy derived from ATP hydrolysis. The mechanistic and structural details regarding PCNA loading by the RFC complex are still developing. In particular, the positive identification of a long-hypothesized structure of an open clamp-RFC complex as an intermediate in loading has remained elusive. In this study, we capture an open yeast PCNA clamp in a complex with RFC through fluorescence energy transfer experiments. We also follow the topological transitions of PCNA in the various steps of the clamp-loading pathway through both steady-state and stopped-flow fluorescence studies. We find that ATP effectively drives the clamp-loading process to completion with the formation of the closed PCNA bound to DNA, whereas ATPgammaS cannot. The information derived from this work complements that obtained from previous structural and mechanistic studies and provides a more complete picture of a eukaryotic clamp-loading pathway using yeast as a paradigm.

  5. Electrostatic Interactions at the Dimer Interface Stabilize the E. coli β Sliding Clamp.

    PubMed

    Purohit, Anirban; England, Jennifer K; Douma, Lauren G; Tondnevis, Farzaneh; Bloom, Linda B; Levitus, Marcia

    2017-08-22

    Sliding clamps are ring-shaped oligomeric proteins that encircle DNA and associate with DNA polymerases for processive DNA replication. The dimeric Escherichia coli β-clamp is closed in solution but must adopt an open conformation to be assembled onto DNA by a clamp loader. To determine what factors contribute to the stability of the dimer interfaces in the closed conformation and how clamp dynamics contribute to formation of the open conformation, we identified conditions that destabilized the dimer and measured the effects of these conditions on clamp dynamics. We characterized the role of electrostatic interactions in stabilizing the β-clamp interface. Increasing salt concentration results in decreased dimer stability and faster subunit dissociation kinetics. The equilibrium dissociation constant of the dimeric clamp varies with salt concentration as predicted by simple charge-screening models, indicating that charged amino acids contribute to the remarkable stability of the interface at physiological salt concentrations. Mutation of a charged residue at the interface (Arg-103) weakens the interface significantly, whereas effects are negligible when a hydrophilic (Ser-109) or a hydrophobic (Ile-305) amino acid is mutated instead. It has been suggested that clamp opening by the clamp loader takes advantage of spontaneous opening-closing fluctuations at the clamp's interface, but our time-resolved fluorescence and fluorescence correlation experiments rule out conformational fluctuations that lead to a significant fraction of open states. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  6. Cryo-EM structure of a helicase loading intermediate containing ORC–Cdc6–Cdt1–MCM2-7 bound to DNA

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

    Sun, Jingchuan; Evrin, Cecile; Samel, Stefan A.

    2013-07-14

    In eukaryotes, the Cdt1-bound replicative helicase core MCM2-7 is loaded onto DNA by the ORC–Cdc6 ATPase to form a prereplicative complex (pre-RC) with an MCM2-7 double hexamer encircling DNA. Using purified components in the presence of ATP-γS, we have captured in vitro an intermediate in pre-RC assembly that contains a complex between the ORC–Cdc6 and Cdt1–MCM2-7 heteroheptamers called the OCCM. Cryo-EM studies of this 14-subunit complex reveal that the two separate heptameric complexes are engaged extensively, with the ORC–Cdc6 N-terminal AAA+ domains latching onto the C-terminal AAA+ motor domains of the MCM2-7 hexamer. The conformation of ORC–Cdc6 undergoes a concertedmore » change into a right-handed spiral with helical symmetry that is identical to that of the DNA double helix. The resulting ORC–Cdc6 helicase loader shows a notable structural similarity to the replication factor C clamp loader, suggesting a conserved mechanism of action.« less

  7. EM structure of a helicase-loader complex depicting a 6:2 binding sub-stoichiometry from Geobacillus kaustophilus HTA426

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

    Lin, Yen-Chen; Naveen, Vankadari; Molecular Cell Biology, Taiwan International Graduate Program, Institute of Molecular Biology, Academia Sinica, and Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan

    During DNA replication, bacterial helicase is recruited as a complex in association with loader proteins to unwind the parental duplex. Previous structural studies have reported saturated 6:6 helicase-loader complexes with different conformations. However, structural information on the sub-stoichiometric conformations of these previously-documented helicase-loader complexes remains elusive. Here, with the aid of single particle electron-microscopy (EM) image reconstruction, we present the Geobacillus kaustophilus HTA426 helicase-loader (DnaC-DnaI) complex with a 6:2 binding stoichiometry in the presence of ATPγS. In the 19 Å resolution EM map, the undistorted and unopened helicase ring holds a robust loader density above the C-terminal RecA-like domain. Meanwhile, themore » path of the central DNA binding channel appears to be obstructed by the reconstructed loader density, implying its potential role as a checkpoint conformation to prevent the loading of immature complex onto DNA. Our data also reveals that the bound nucleotides and the consequently induced conformational changes in the helicase hexamer are essential for active association with loader proteins. These observations provide fundamental insights into the formation of the helicase-loader complex in bacteria that regulates the DNA replication process. - Highlights: • Helicase-loader complex structure with 6:2 sub-stoichiometry is resolved by EM. • Helicase hexamer in 6:2 sub-stoichiometry is constricted and un-opened. • 6:2 binding ratio of helicase-loader complex could act as a DNA loading checkpoint. • Nucleotides stabilize helicase-loader complex at low protein concentrations.« less

  8. Structure and Biochemical Activities of Escherichia coli MgsA

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

    Page, Asher N.; George, Nicholas P.; Marceau, Aimee H.

    2012-02-27

    Bacterial 'maintenance of genome stability protein A' (MgsA) and related eukaryotic enzymes play important roles in cellular responses to stalled DNA replication processes. Sequence information identifies MgsA enzymes as members of the clamp loader clade of AAA{sup +} proteins, but structural information defining the family has been limited. Here, the x-ray crystal structure of Escherichia coli MgsA is described, revealing a homotetrameric arrangement for the protein that distinguishes it from other clamp loader clade AAA{sup +} proteins. Each MgsA protomer is composed of three elements as follows: ATP-binding and helical lid domains (conserved among AAA{sup +} proteins) and a tetramerizationmore » domain. Although the tetramerization domains bury the greatest amount of surface area in the MgsA oligomer, each of the domains participates in oligomerization to form a highly intertwined quaternary structure. Phosphate is bound at each AAA{sup +} ATP-binding site, but the active sites do not appear to be in a catalytically competent conformation due to displacement of Arg finger residues. E. coli MgsA is also shown to form a complex with the single-stranded DNA-binding protein through co-purification and biochemical studies. MgsA DNA-dependent ATPase activity is inhibited by single-stranded DNA-binding protein. Together, these structural and biochemical observations provide insights into the mechanisms of MgsA family AAA{sup +} proteins.« less

  9. Structure and Biochemical Activities of Escherichia coli MgsA*♦

    PubMed Central

    Page, Asher N.; George, Nicholas P.; Marceau, Aimee H.; Cox, Michael M.; Keck, James L.

    2011-01-01

    Bacterial “maintenance of genome stability protein A” (MgsA) and related eukaryotic enzymes play important roles in cellular responses to stalled DNA replication processes. Sequence information identifies MgsA enzymes as members of the clamp loader clade of AAA+ proteins, but structural information defining the family has been limited. Here, the x-ray crystal structure of Escherichia coli MgsA is described, revealing a homotetrameric arrangement for the protein that distinguishes it from other clamp loader clade AAA+ proteins. Each MgsA protomer is composed of three elements as follows: ATP-binding and helical lid domains (conserved among AAA+ proteins) and a tetramerization domain. Although the tetramerization domains bury the greatest amount of surface area in the MgsA oligomer, each of the domains participates in oligomerization to form a highly intertwined quaternary structure. Phosphate is bound at each AAA+ ATP-binding site, but the active sites do not appear to be in a catalytically competent conformation due to displacement of Arg finger residues. E. coli MgsA is also shown to form a complex with the single-stranded DNA-binding protein through co-purification and biochemical studies. MgsA DNA-dependent ATPase activity is inhibited by single-stranded DNA-binding protein. Together, these structural and biochemical observations provide insights into the mechanisms of MgsA family AAA+ proteins. PMID:21297161

  10. Single-molecule visualization of fast polymerase turnover in the bacterial replisome

    PubMed Central

    Lewis, Jacob S; Spenkelink, Lisanne M; Jergic, Slobodan; Wood, Elizabeth A; Monachino, Enrico; Horan, Nicholas P; Duderstadt, Karl E; Cox, Michael M; Robinson, Andrew; Dixon, Nicholas E; van Oijen, Antoine M

    2017-01-01

    The Escherichia coli DNA replication machinery has been used as a road map to uncover design rules that enable DNA duplication with high efficiency and fidelity. Although the enzymatic activities of the replicative DNA Pol III are well understood, its dynamics within the replisome are not. Here, we test the accepted view that the Pol III holoenzyme remains stably associated within the replisome. We use in vitro single-molecule assays with fluorescently labeled polymerases to demonstrate that the Pol III* complex (holoenzyme lacking the β2 sliding clamp), is rapidly exchanged during processive DNA replication. Nevertheless, the replisome is highly resistant to dilution in the absence of Pol III* in solution. We further show similar exchange in live cells containing labeled clamp loader and polymerase. These observations suggest a concentration-dependent exchange mechanism providing a balance between stability and plasticity, facilitating replacement of replisomal components dependent on their availability in the environment. DOI: http://dx.doi.org/10.7554/eLife.23932.001 PMID:28432790

  11. Investigation of sliding DNA clamp dynamics by single-molecule fluorescence, mass spectrometry and structure-based modeling

    PubMed Central

    Gadkari, Varun V; Harvey, Sophie R; Raper, Austin T; Chu, Wen-Ting; Wang, Jin; Wysocki, Vicki H; Suo, Zucai

    2018-01-01

    Abstract Proliferating cell nuclear antigen (PCNA) is a trimeric ring-shaped clamp protein that encircles DNA and interacts with many proteins involved in DNA replication and repair. Despite extensive structural work to characterize the monomeric, dimeric, and trimeric forms of PCNA alone and in complex with interacting proteins, no structure of PCNA in a ring-open conformation has been published. Here, we use a multidisciplinary approach, including single-molecule Förster resonance energy transfer (smFRET), native ion mobility-mass spectrometry (IM-MS), and structure-based computational modeling, to explore the conformational dynamics of a model PCNA from Sulfolobus solfataricus (Sso), an archaeon. We found that Sso PCNA samples ring-open and ring-closed conformations even in the absence of its clamp loader complex, replication factor C, and transition to the ring-open conformation is modulated by the ionic strength of the solution. The IM-MS results corroborate the smFRET findings suggesting that PCNA dynamics are maintained in the gas phase and further establishing IM-MS as a reliable strategy to investigate macromolecular motions. Our molecular dynamic simulations agree with the experimental data and reveal that ring-open PCNA often adopts an out-of-plane left-hand geometry. Collectively, these results implore future studies to define the roles of PCNA dynamics in DNA loading and other PCNA-mediated interactions. PMID:29529283

  12. Structure of the active form of human origin recognition complex and its ATPase motor module

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

    Tocilj, Ante; On, Kin Fan; Yuan, Zuanning

    Binding of the Origin Recognition Complex (ORC) to origins of replication marks the first step in the initiation of replication of the genome in all eukaryotic cells. Here, we report the structure of the active form of human ORC determined by X-ray crystallography and cryo-electron microscopy. The complex is composed of an ORC1/4/5 motor module lobe in an organization reminiscent of the DNA polymerase clamp loader complexes. A second lobe contains the ORC2/3 subunits. The complex is organized as a double-layered shallow corkscrew, with the AAA+ and AAA+-like domains forming one layer, and the winged-helix domains (WHDs) forming a topmore » layer. CDC6 fits easily between ORC1 and ORC2, completing the ring and the DNA-binding channel, forming an additional ATP hydrolysis site. Analysis of the ATPase activity of the complex provides a basis for understanding ORC activity as well as molecular defects observed in Meier-Gorlin Syndrome mutations.« less

  13. Structure of a helicase–helicase loader complex reveals insights into the mechanism of bacterial primosome assembly

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

    Liu, Bin; Eliason, William K.; Steitz, Thomas A.

    2013-09-19

    During the assembly of the bacterial loader-dependent primosome, helicase loader proteins bind to the hexameric helicase ring, deliver it onto the oriC DNA and then dissociate from the complex. Here, to provide a better understanding of this key process, we report the crystal structure of the ~570-kDa prepriming complex between the Bacillus subtilis loader protein and the Bacillus stearothermophilus helicase, as well as the helicase-binding domain of primase with a molar ratio of 6:6:3 at 7.5 Å resolution. The overall architecture of the complex exhibits a three-layered ring conformation. Moreover, the structure combined with the proposed model suggests that themore » shift from the ‘open-ring’ to the ‘open-spiral’ and then the ‘closed-spiral’ state of the helicase ring due to the binding of single-stranded DNA may be the cause of the loader release.« less

  14. Structure of the active form of human origin recognition complex and its ATPase motor module

    PubMed Central

    Tocilj, Ante; On, Kin Fan; Yuan, Zuanning; Sun, Jingchuan; Elkayam, Elad; Li, Huilin; Stillman, Bruce; Joshua-Tor, Leemor

    2017-01-01

    Binding of the Origin Recognition Complex (ORC) to origins of replication marks the first step in the initiation of replication of the genome in all eukaryotic cells. Here, we report the structure of the active form of human ORC determined by X-ray crystallography and cryo-electron microscopy. The complex is composed of an ORC1/4/5 motor module lobe in an organization reminiscent of the DNA polymerase clamp loader complexes. A second lobe contains the ORC2/3 subunits. The complex is organized as a double-layered shallow corkscrew, with the AAA+ and AAA+-like domains forming one layer, and the winged-helix domains (WHDs) forming a top layer. CDC6 fits easily between ORC1 and ORC2, completing the ring and the DNA-binding channel, forming an additional ATP hydrolysis site. Analysis of the ATPase activity of the complex provides a basis for understanding ORC activity as well as molecular defects observed in Meier-Gorlin Syndrome mutations. DOI: http://dx.doi.org/10.7554/eLife.20818.001 PMID:28112645

  15. Roles of the checkpoint sensor clamp Rad9-Rad1-Hus1 (911)-complex and the clamp loaders Rad17-RFC and Ctf18-RFC in Schizosaccharomyces pombe telomere maintenance.

    PubMed

    Khair, Lyne; Chang, Ya-Ting; Subramanian, Lakxmi; Russell, Paul; Nakamura, Toru M

    2010-06-01

    While telomeres must provide mechanisms to prevent DNA repair and DNA damage checkpoint factors from fusing chromosome ends and causing permanent cell cycle arrest, these factors associate with functional telomeres and play critical roles in the maintenance of telomeres. Previous studies have established that Tel1 (ATM) and Rad3 (ATR) kinases play redundant but essential roles for telomere maintenance in fission yeast. In addition, the Rad9-Rad1-Hus1 (911) and Rad17-RFC complexes work downstream of Rad3 (ATR) in fission yeast telomere maintenance. Here, we investigated how 911, Rad17-RFC and another RFC-like complex Ctf18-RFC contribute to telomere maintenance in fission yeast cells lacking Tel1 and carrying a novel hypomorphic allele of rad3 (DBD-rad3), generated by the fusion between the DNA binding domain (DBD) of the fission yeast telomere capping protein Pot1 and Rad3. Our investigations have uncovered a surprising redundancy for Rad9 and Hus1 in allowing Rad1 to contribute to telomere maintenance in DBD-rad3 tel1 cells. In addition, we found that Rad17-RFC and Ctf18-RFC carry out redundant telomere maintenance functions in DBD-rad3 tel1 cells. Since checkpoint sensor proteins are highly conserved, genetic redundancies uncovered here may be relevant to telomere maintenance and detection of DNA damage in other eukaryotes.

  16. Roles of the Checkpoint Sensor Clamp Rad9-Rad1-Hus1 (911)-Complex and the Clamp Loaders Rad17-RFC and Ctf18-RFC in Schizosaccharomyces pombe Telomere Maintenance

    PubMed Central

    Khair, Lyne; Chang, Ya-Ting; Subramanian, Lakxmi; Russell, Paul; Nakamura, Toru M.

    2011-01-01

    While telomeres must provide mechanisms to prevent DNA repair and DNA damage checkpoint factors from fusing chromosome ends and causing permanent cell cycle arrest, these factors associate with functional telomeres and play critical roles in the maintenance of telomeres. Previous studies have established that Tel1 (ATM) and Rad3 (ATR) kinases play redundant but essential roles for telomere maintenance in fission yeast. In addition, the Rad9-Rad1-Hus1 (911) and Rad17-RFC complexes work downstream of Rad3 (ATR) in fission yeast telomere maintenance. Here, we investigated how 911, Rad17-RFC and another RFC-like complex Ctf18-RFC contribute to telomere maintenance in fission yeast cells lacking Tel1 and carrying a novel hypomorphic allele of rad3 (DBD-rad3), generated by the fusion between the DNA binding domain (DBD) of the fission yeast telomere capping protein Pot1 and Rad3. Our investigations have uncovered a surprising redundancy for Rad9 and Hus1 in allowing Rad1 to contribute to telomere maintenance in DBD-rad3 tel1Δ cells. In addition, we found that Rad17-RFC and Ctf18-RFC carry out redundant telomere maintenance functions in DBD-rad3 tel1Δ cells. Since checkpoint sensor proteins are highly conserved, genetic redundancies uncovered here may be relevant to telomere maintenance and detection of DNA damage in other eukaryotes. PMID:20505337

  17. The Ctf18RFC Clamp Loader Is Essential for Telomere Stability in Telomerase-Negative and mre11 Mutant Alleles

    PubMed Central

    Parke, Courtney; Tatum, Danielle; Lustig, Arthur J.

    2014-01-01

    The function of the replication clamp loaders in the semi-conservative telomere replication and their relationship to telomerase- and recombination mechanisms of telomere addition remains ambiguous. We have investigated the variant clamp loader Ctf18 RFC (Replication Factor C). To understand the role of Ctf18 at the telomere, we first investigated genetic interactions after loss of Ctf18 and TLC1 (the yeast telomerase RNA). We find that the tlc1▵ ctf18▵ double mutant confers a rapid >1000-fold decrease in viability. The rate of loss was similar to the kinetics of cell death in rad52▵ tlc1▵ cells. However, the Ctf18 pathway is distinct from Rad52, required for the repair of DSBs, as demonstrated by the synthetic lethality of rad52▵ tlc1▵ ctf18▵ triple mutants. These data suggest that each mutant elicits non-redundant defects acting on the same substrate. Second, interactions of the yeast hyper-recombinational mutant, mre11A470T, with ctf18▵ confer a synergistic cold sensitivity. The phenotype of these double mutants ultimately results in telomere loss and the generation of recombinational survivors. We observed a similar synergism between single mutants that led to hypersensitivity to the DNA alkylating agent, methane methyl sulphonate (MMS), the replication fork inhibitor hydroxyurea (HU), and to a failure to separate telomeres of sister chromatids. Hence, ctf18▵ and mre11A470T act in different pathways on telomere substrates for multiple phenotypes. The mre11A470T cells also displayed a DNA damage response (DDR) at 15°C but not at 30°C while ctf18▵ mutants conferred a constitutive DDR activity. Both the 15°C DDR pattern and growth rate were reversible at 30°C and displayed telomerase activity in vivo. We hypothesize that Ctf18 confers protection against stalling and/or breaks at the replication fork in cells that either lack, or are compromised for, telomerase activity. This Ctf18-based function is likely to contribute another level to telomere size homeostasis. PMID:24533124

  18. The causes and control of loader- and truck-related fatalities in surface mining operations.

    PubMed

    Kecojevic, Vladislav; Radomsky, Mark

    2004-12-01

    At surface mining operations throughout the world, loaders and trucks are a primary means of material loading and haulage. As the size, use and technological complexity of these units have increased, so has the concern regarding loader and truck safety. The severity and number of accidents involving loaders and trucks is higher when compared to all other mining accident types. In this paper, an analysis of loader and truck-related fatalities over the last 8 years is performed, the fatality categories and causes of accidents are established and control strategies are discussed and evaluated in an effort to increase hazard awareness by emphasizing safe loading, hauling and maintenance practices, as well as the value of traditional and innovative miner training programmes.

  19. The logic of DNA replication in double-stranded DNA viruses: insights from global analysis of viral genomes

    PubMed Central

    Kazlauskas, Darius; Krupovic, Mart; Venclovas, Česlovas

    2016-01-01

    Abstract Genomic DNA replication is a complex process that involves multiple proteins. Cellular DNA replication systems are broadly classified into only two types, bacterial and archaeo-eukaryotic. In contrast, double-stranded (ds) DNA viruses feature a much broader diversity of DNA replication machineries. Viruses differ greatly in both completeness and composition of their sets of DNA replication proteins. In this study, we explored whether there are common patterns underlying this extreme diversity. We identified and analyzed all major functional groups of DNA replication proteins in all available proteomes of dsDNA viruses. Our results show that some proteins are common to viruses infecting all domains of life and likely represent components of the ancestral core set. These include B-family polymerases, SF3 helicases, archaeo-eukaryotic primases, clamps and clamp loaders of the archaeo-eukaryotic type, RNase H and ATP-dependent DNA ligases. We also discovered a clear correlation between genome size and self-sufficiency of viral DNA replication, the unanticipated dominance of replicative helicases and pervasive functional associations among certain groups of DNA replication proteins. Altogether, our results provide a comprehensive view on the diversity and evolution of replication systems in the DNA virome and uncover fundamental principles underlying the orchestration of viral DNA replication. PMID:27112572

  20. Single-molecule visualization of Saccharomyces cerevisiae leading-strand synthesis reveals dynamic interaction between MTC and the replisome

    PubMed Central

    Lewis, Jacob S.; Spenkelink, Lisanne M.; Schauer, Grant D.; Hill, Flynn R.; Georgescu, Roxanna E.; O’Donnell, Michael E.; van Oijen, Antoine M.

    2017-01-01

    The replisome, the multiprotein system responsible for genome duplication, is a highly dynamic complex displaying a large number of different enzyme activities. Recently, the Saccharomyces cerevisiae minimal replication reaction has been successfully reconstituted in vitro. This provided an opportunity to uncover the enzymatic activities of many of the components in a eukaryotic system. Their dynamic behavior and interactions in the context of the replisome, however, remain unclear. We use a tethered-bead assay to provide real-time visualization of leading-strand synthesis by the S. cerevisiae replisome at the single-molecule level. The minimal reconstituted leading-strand replisome requires 24 proteins, forming the CMG helicase, the Pol ε DNA polymerase, the RFC clamp loader, the PCNA sliding clamp, and the RPA single-stranded DNA binding protein. We observe rates and product lengths similar to those obtained from ensemble biochemical experiments. At the single-molecule level, we probe the behavior of two components of the replication progression complex and characterize their interaction with active leading-strand replisomes. The Minichromosome maintenance protein 10 (Mcm10), an important player in CMG activation, increases the number of productive replication events in our assay. Furthermore, we show that the fork protection complex Mrc1–Tof1–Csm3 (MTC) enhances the rate of the leading-strand replisome threefold. The introduction of periods of fast replication by MTC leads to an average rate enhancement of a factor of 2, similar to observations in cellular studies. We observe that the MTC complex acts in a dynamic fashion with the moving replisome, leading to alternating phases of slow and fast replication. PMID:28923950

  1. 146. FUEL LINE TO SKID 2 (FUEL LOADER) IN FUEL ...

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

    146. FUEL LINE TO SKID 2 (FUEL LOADER) IN FUEL CONTROL ROOM (215), LSB (BLDG. 751). LIQUID NITROGEN/HELIUM HEAT EXCHANGER ON RIGHT. - Vandenberg Air Force Base, Space Launch Complex 3, Launch Pad 3 East, Napa & Alden Roads, Lompoc, Santa Barbara County, CA

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

    Hood, Iris V.; Berger, James M.

    Replisome assembly requires the loading of replicative hexameric helicases onto origins by AAA+ ATPases. How loader activity is appropriately controlled remains unclear. Here, we use structural and biochemical analyses to establish how an antimicrobial phage protein interferes with the function of theStaphylococcus aureusreplicative helicase loader, DnaI. The viral protein binds to the loader’s AAA+ ATPase domain, allowing binding of the host replicative helicase but impeding loader self-assembly and ATPase activity. Close inspection of the complex highlights an unexpected locus for the binding of an interdomain linker element in DnaI/DnaC-family proteins. We find that the inhibitor protein is genetically coupled tomore » a phage-encoded homolog of the bacterial helicase loader, which we show binds to the host helicase but not to the inhibitor itself. These findings establish a new approach by which viruses can hijack host replication processes and explain how loader activity is internally regulated to prevent aberrant auto-association.« less

  3. Ontology-Based Method for Fault Diagnosis of Loaders.

    PubMed

    Xu, Feixiang; Liu, Xinhui; Chen, Wei; Zhou, Chen; Cao, Bingwei

    2018-02-28

    This paper proposes an ontology-based fault diagnosis method which overcomes the difficulty of understanding complex fault diagnosis knowledge of loaders and offers a universal approach for fault diagnosis of all loaders. This method contains the following components: (1) An ontology-based fault diagnosis model is proposed to achieve the integrating, sharing and reusing of fault diagnosis knowledge for loaders; (2) combined with ontology, CBR (case-based reasoning) is introduced to realize effective and accurate fault diagnoses following four steps (feature selection, case-retrieval, case-matching and case-updating); and (3) in order to cover the shortages of the CBR method due to the lack of concerned cases, ontology based RBR (rule-based reasoning) is put forward through building SWRL (Semantic Web Rule Language) rules. An application program is also developed to implement the above methods to assist in finding the fault causes, fault locations and maintenance measures of loaders. In addition, the program is validated through analyzing a case study.

  4. Ontology-Based Method for Fault Diagnosis of Loaders

    PubMed Central

    Liu, Xinhui; Chen, Wei; Zhou, Chen; Cao, Bingwei

    2018-01-01

    This paper proposes an ontology-based fault diagnosis method which overcomes the difficulty of understanding complex fault diagnosis knowledge of loaders and offers a universal approach for fault diagnosis of all loaders. This method contains the following components: (1) An ontology-based fault diagnosis model is proposed to achieve the integrating, sharing and reusing of fault diagnosis knowledge for loaders; (2) combined with ontology, CBR (case-based reasoning) is introduced to realize effective and accurate fault diagnoses following four steps (feature selection, case-retrieval, case-matching and case-updating); and (3) in order to cover the shortages of the CBR method due to the lack of concerned cases, ontology based RBR (rule-based reasoning) is put forward through building SWRL (Semantic Web Rule Language) rules. An application program is also developed to implement the above methods to assist in finding the fault causes, fault locations and maintenance measures of loaders. In addition, the program is validated through analyzing a case study. PMID:29495646

  5. The logic of DNA replication in double-stranded DNA viruses: insights from global analysis of viral genomes.

    PubMed

    Kazlauskas, Darius; Krupovic, Mart; Venclovas, Česlovas

    2016-06-02

    Genomic DNA replication is a complex process that involves multiple proteins. Cellular DNA replication systems are broadly classified into only two types, bacterial and archaeo-eukaryotic. In contrast, double-stranded (ds) DNA viruses feature a much broader diversity of DNA replication machineries. Viruses differ greatly in both completeness and composition of their sets of DNA replication proteins. In this study, we explored whether there are common patterns underlying this extreme diversity. We identified and analyzed all major functional groups of DNA replication proteins in all available proteomes of dsDNA viruses. Our results show that some proteins are common to viruses infecting all domains of life and likely represent components of the ancestral core set. These include B-family polymerases, SF3 helicases, archaeo-eukaryotic primases, clamps and clamp loaders of the archaeo-eukaryotic type, RNase H and ATP-dependent DNA ligases. We also discovered a clear correlation between genome size and self-sufficiency of viral DNA replication, the unanticipated dominance of replicative helicases and pervasive functional associations among certain groups of DNA replication proteins. Altogether, our results provide a comprehensive view on the diversity and evolution of replication systems in the DNA virome and uncover fundamental principles underlying the orchestration of viral DNA replication. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  6. Multi-Sensor Based Online Attitude Estimation and Stability Measurement of Articulated Heavy Vehicles.

    PubMed

    Zhu, Qingyuan; Xiao, Chunsheng; Hu, Huosheng; Liu, Yuanhui; Wu, Jinjin

    2018-01-13

    Articulated wheel loaders used in the construction industry are heavy vehicles and have poor stability and a high rate of accidents because of the unpredictable changes of their body posture, mass and centroid position in complex operation environments. This paper presents a novel distributed multi-sensor system for real-time attitude estimation and stability measurement of articulated wheel loaders to improve their safety and stability. Four attitude and heading reference systems (AHRS) are constructed using micro-electro-mechanical system (MEMS) sensors, and installed on the front body, rear body, rear axis and boom of an articulated wheel loader to detect its attitude. A complementary filtering algorithm is deployed for sensor data fusion in the system so that steady state margin angle (SSMA) can be measured in real time and used as the judge index of rollover stability. Experiments are conducted on a prototype wheel loader, and results show that the proposed multi-sensor system is able to detect potential unstable states of an articulated wheel loader in real-time and with high accuracy.

  7. Multi-Sensor Based Online Attitude Estimation and Stability Measurement of Articulated Heavy Vehicles

    PubMed Central

    Xiao, Chunsheng; Liu, Yuanhui; Wu, Jinjin

    2018-01-01

    Articulated wheel loaders used in the construction industry are heavy vehicles and have poor stability and a high rate of accidents because of the unpredictable changes of their body posture, mass and centroid position in complex operation environments. This paper presents a novel distributed multi-sensor system for real-time attitude estimation and stability measurement of articulated wheel loaders to improve their safety and stability. Four attitude and heading reference systems (AHRS) are constructed using micro-electro-mechanical system (MEMS) sensors, and installed on the front body, rear body, rear axis and boom of an articulated wheel loader to detect its attitude. A complementary filtering algorithm is deployed for sensor data fusion in the system so that steady state margin angle (SSMA) can be measured in real time and used as the judge index of rollover stability. Experiments are conducted on a prototype wheel loader, and results show that the proposed multi-sensor system is able to detect potential unstable states of an articulated wheel loader in real-time and with high accuracy. PMID:29342850

  8. A 3 kbar hydrogen-compatible gas loader for Paris-Edinburgh presses

    NASA Astrophysics Data System (ADS)

    Klotz, S.; Philippe, J.; Bull, C. L.; Loveday, J. S.; Nelmes, R. J.

    2013-03-01

    We present a device which allows compressed gases to be loaded into large volume opposed anvils used for high pressure neutron scattering in the multi-10 GPa range. The gases are initially loaded into clamps which can then be inserted into VX-Paris-Edinburgh load frames. The system is compatible with all inert gases as well as hydrogen and permits loading pressures of up to 3 kbar for which most gases have densities close to that of the liquid at ambient pressure. The device should have applications for the study of simple molecular solids as well as for loading gases as pressure-transmitting media.

  9. Sliding Clamp–DNA Interactions Are Required for Viability and Contribute to DNA Polymerase Management in Escherichia coli

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

    Heltzel, J.; Scouten Ponticelli, S; Sanders, L

    2009-01-01

    Sliding clamp proteins topologically encircle DNA and play vital roles in coordinating the actions of various DNA replication, repair, and damage tolerance proteins. At least three distinct surfaces of the Escherichia coli {beta} clamp interact physically with the DNA that it topologically encircles. We utilized mutant {beta} clamp proteins bearing G66E and G174A substitutions ({beta}159), affecting the single-stranded DNA-binding region, or poly-Ala substitutions in place of residues 148-HQDVR-152 ({beta}148-152), affecting the double-stranded DNA binding region, to determine the biological relevance of clamp-DNA interactions. As part of this work, we solved the X-ray crystal structure of {beta}148-152, which verified that themore » poly-Ala substitutions failed to significantly alter the tertiary structure of the clamp. Based on functional assays, both {beta}159 and {beta}148-152 were impaired for loading and retention on a linear primed DNA in vitro. In the case of {beta}148-152, this defect was not due to altered interactions with the DnaX clamp loader, but rather was the result of impaired {beta}148-152-DNA interactions. Once loaded, {beta}148-152 was proficient for DNA polymerase III (Pol III) replication in vitro. In contrast, {beta}148-152 was severely impaired for Pol II and Pol IV replication and was similarly impaired for direct physical interactions with these Pols. Despite its ability to support Pol III replication in vitro, {beta}148-152 was unable to support viability of E. coli. Nevertheless, physiological levels of {beta}148-152 expressed from a plasmid efficiently complemented the temperature-sensitive growth phenotype of a strain expressing {beta}159 (dnaN159), provided that Pol II and Pol IV were inactivated. Although this strain was impaired for Pol V-dependent mutagenesis, inactivation of Pol II and Pol IV restored the Pol V mutator phenotype. Taken together, these results support a model in which a sophisticated combination of competitive clamp-DNA, clamp-partner, and partner-DNA interactions serve to manage the actions of the different E. coli Pols in vivo.« less

  10. 1. MAGAZINE COMPLEX, O, P, AND Q. MAGAZINE P IN ...

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

    1. MAGAZINE COMPLEX, O, P, AND Q. MAGAZINE P IN BACKGROUND, LOOKING NORTHEAST. - NIKE Missile Base C-84, Underground Storage Magazines & Launcher-Loader Assemblies, Easternmost portion of launch area, Barrington, Cook County, IL

  11. The effects of a dynamic graphical model during simulation-based training of console operation skill

    NASA Technical Reports Server (NTRS)

    Farquhar, John D.; Regian, J. Wesley

    1993-01-01

    LOADER is a Windows-based simulation of a complex procedural task. The task requires subjects to execute long sequences of console-operation actions (e.g., button presses, switch actuations, dial rotations) to accomplish specific goals. The LOADER interface is a graphical computer-simulated console which controls railroad cars, tracks, and cranes in a fictitious railroad yard. We hypothesized that acquisition of LOADER performance skill would be supported by the representation of a dynamic graphical model linking console actions to goal and goal states in the 'railroad yard'. Twenty-nine subjects were randomly assigned to one of two treatments (i.e., dynamic model or no model). During training, both groups received identical text-based instruction in an instructional-window above the LOADER interface. One group, however, additionally saw a dynamic version of the bird's-eye view of the railroad yard. After training, both groups were tested under identical conditions. They were asked to perform the complete procedure without guidance and without access to either type of railroad yard representation. Results indicate that rather than becoming dependent on the animated rail yard model, subjects in the dynamic model condition apparently internalized the model, as evidenced by their performance after the model was removed.

  12. An isolated Hda-clamp complex is functional in the regulatory inactivation of DnaA and DNA replication.

    PubMed

    Kawakami, Hironori; Su'etsugu, Masayuki; Katayama, Tsutomu

    2006-10-01

    In Escherichia coli, a complex consisting of Hda and the DNA-loaded clamp-subunit of the DNA polymerase III holoenzyme promotes hydrolysis of DnaA-ATP. The resultant ADP-DnaA is inactive for initiation of chromosomal DNA replication, thereby repressing excessive initiations. As the cellular content of the clamp is 10-100 times higher than that of Hda, most Hda molecules might be complexed with the clamp in vivo. Although Hda predominantly forms irregular aggregates when overexpressed, in the present study we found that co-overexpression of the clamp with Hda enhances Hda solubility dramatically and we efficiently isolated the Hda-clamp complex. A single molecule of the complex appears to consist of two Hda molecules and a single clamp. The complex is competent in DnaA-ATP hydrolysis and DNA replication in the presence of DNA and the clamp deficient subassembly of the DNA polymerase III holoenzyme (pol III*). These findings indicate that the clamp contained in the complex is loaded onto DNA through an interaction with the pol III* and that the Hda activity is preserved in these processes. The complex consisting of Hda and the DNA-unloaded clamp may play a specific role in a process proceeding to the DnaA-ATP hydrolysis in vivo.

  13. Wheel loader round up

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

    Fiscor, S.

    2007-02-15

    Operator comfort and serviceability remain important features of front-end loaders or wheel loaders. The article reports on features of the latest large loaders from three companies: Caterpillar (the 992G and 994F), Komatsu (the WA800-3, WA900-3 and WA1200-3), and Le Tourneau (the L-950, L-1350, L-1850 and L-2350). 1 tab., 4 photos.

  14. Seismic explosive charge loader and anchor

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

    Mcreynolds, O.B.

    1981-07-14

    An improved seismic explosive charge loader and anchor for loading and anchoring explosives in cylindrical containers in bore holes is disclosed, which includes a snap in spring band shaped anchor which effectively anchors the loader in the well bore against upward movement, one aspect of the invention includes a snap lock threaded connection for securing an explosive container having interrupted threads to the loader and anchor, and the loader and anchor is constructed and arranged to maintain a detonator in place in the explosive container thereby assuring detonation of the explosive.

  15. The RNA polymerase clamp interconverts dynamically among three states and is stabilized in a partly closed state by ppGpp.

    PubMed

    Duchi, Diego; Mazumder, Abhishek; Malinen, Anssi M; Ebright, Richard H; Kapanidis, Achillefs N

    2018-06-06

    RNA polymerase (RNAP) contains a mobile structural module, the 'clamp,' that forms one wall of the RNAP active-center cleft and that has been linked to crucial aspects of the transcription cycle, including promoter melting, transcription elongation complex stability, transcription pausing, and transcription termination. Using single-molecule FRET on surface-immobilized RNAP molecules, we show that the clamp in RNAP holoenzyme populates three distinct conformational states and interconvert between these states on the 0.1-1 s time-scale. Similar studies confirm that the RNAP clamp is closed in open complex (RPO) and in initial transcribing complexes (RPITC), including paused initial transcribing complexes, and show that, in these complexes, the clamp does not exhibit dynamic behaviour. We also show that, the stringent-response alarmone ppGpp, which reprograms transcription during amino acid starvation stress, selectively stabilizes the partly-closed-clamp state and prevents clamp opening; these results raise the possibility that ppGpp controls promoter opening by modulating clamp dynamics.

  16. Extracting whole short rotation trees with a skidder and a front-end loader

    Treesearch

    R. Spinelli; B.R. Hartsough

    2001-01-01

    We time-studied a Caterpillar 950F front-end loader and a Caterpillar 528 grapple skidder used to extract bunched whole trees to a landing in a short rotation Eucalyptus plantation. The loader was 40-60% more productive than the grapple skidder, depending on extraction distance. Alternatively, the single loader could both extract trees and handle the landing duties,...

  17. EPA Nonregulatory Nonroad Duty Cycles

    EPA Pesticide Factsheets

    EPA nonregulatory, nonroad duty cycles for equipment such as agricultural tractors, backhoe loaders,crawlers tractors, excavators, arc welding skid steer loaders, and wheel loaders. Also,test procedures, laboratory methods, and emissions for this equipmen

  18. Development of an electric, battery powered, skid-steer loader

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

    Alcock, R.; Froehlich, D.P.; Christianson, L.L.

    1985-01-01

    A battery powered, skid-steer loader was developed for farm chore routines and materials handling activities. Outlined are details on drive train, hydraulics, loader, frame, cab, controls, battery and controller. Preliminary performance characteristics of the hydraulics are presented.

  19. Automatic Vetting for Malice in Android Platforms

    DTIC Science & Technology

    2016-05-01

    be distinguished. For example, both Face- book and Instagram ship a class named Landroid/support/v4/app/Fragment;§ with different method signatures...relationships (e.g., Lcom/ instagram /.../LoadImageTask; is a sub- class of the abstract class Landroid/os/AsyncTask;). Figure 2(c) displays a method graph...7feaf7c75a5305b1083a160f...baa6.dex.dex parent_loader 3 Instagram instagram_classes.dex parent_loader 4 JohnNESLite johnneslite_classes.dex parent_loader (a) Class Loader

  20. Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing.

    PubMed

    Kim, Jin S; Nanfara, Michael T; Chodavarapu, Sundari; Jin, Kyeong S; Babu, Vignesh M P; Ghazy, Mohamed A; Chung, Scisung; Kaguni, Jon M; Sutton, Mark D; Cho, Yunje

    2017-04-20

    Regulatory inactivation of DnaA (RIDA) is one of the major regulatory mechanisms of prokaryotic replication licensing. In RIDA, the Hda-sliding clamp complex loaded onto DNA directly interacts with adenosine triphosphate (ATP)-bound DnaA and stimulates the hydrolysis of ATP to inactivate DnaA. A prediction is that the activity of Hda is tightly controlled to ensure that replication initiation occurs only once per cell cycle. Here, we determined the crystal structure of the Hda-β clamp complex. This complex contains two pairs of Hda dimers sandwiched between two β clamp rings to form an octamer that is stabilized by three discrete interfaces. Two separate surfaces of Hda make contact with the β clamp, which is essential for Hda function in RIDA. The third interface between Hda monomers occludes the active site arginine finger, blocking its access to DnaA. Taken together, our structural and mutational analyses of the Hda-β clamp complex indicate that the interaction of the β clamp with Hda controls the ability of Hda to interact with DnaA. In the octameric Hda-β clamp complex, the inability of Hda to interact with DnaA is a novel mechanism that may regulate Hda function. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  1. Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing

    PubMed Central

    Kim, Jin S.; Nanfara, Michael T.; Chodavarapu, Sundari; Jin, Kyeong S.; Babu, Vignesh M. P.; Ghazy, Mohamed A.; Chung, Scisung

    2017-01-01

    Abstract Regulatory inactivation of DnaA (RIDA) is one of the major regulatory mechanisms of prokaryotic replication licensing. In RIDA, the Hda–sliding clamp complex loaded onto DNA directly interacts with adenosine triphosphate (ATP)-bound DnaA and stimulates the hydrolysis of ATP to inactivate DnaA. A prediction is that the activity of Hda is tightly controlled to ensure that replication initiation occurs only once per cell cycle. Here, we determined the crystal structure of the Hda–β clamp complex. This complex contains two pairs of Hda dimers sandwiched between two β clamp rings to form an octamer that is stabilized by three discrete interfaces. Two separate surfaces of Hda make contact with the β clamp, which is essential for Hda function in RIDA. The third interface between Hda monomers occludes the active site arginine finger, blocking its access to DnaA. Taken together, our structural and mutational analyses of the Hda–β clamp complex indicate that the interaction of the β clamp with Hda controls the ability of Hda to interact with DnaA. In the octameric Hda–β clamp complex, the inability of Hda to interact with DnaA is a novel mechanism that may regulate Hda function. PMID:28168278

  2. Compilation of load spectrum of loader drive axle

    NASA Astrophysics Data System (ADS)

    Wei, Yongxiang; Zhu, Haoyue; Tang, Heng; Yuan, Qunwei

    2018-03-01

    In order to study the preparation method of gear fatigue load spectrum for loaders, the load signal of four typical working conditions of loader is collected. The signal that reflects the law of load change is obtained by preprocessing the original signal. The torque of the drive axle is calculated by using the rain flow counting method. According to the operating time ratio of each working condition, the two dimensional load spectrum based on the real working conditions of the drive axle of loader is established by the cycle extrapolation and synthesis method. The two-dimensional load spectrum is converted into one-dimensional load spectrum by means of the mean of torque equal damage method. Torque amplification includes the maximum load torque of the main reduction gear. Based on the theory of equal damage, the accelerated cycles are calculated. In this way, the load spectrum of the loading condition of the drive axle is prepared to reflect loading condition of the loader. The load spectrum can provide reference for fatigue life test and life prediction of loader drive axle.

  3. Mound powder loader, Mod 1

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

    Gress, A.V. Jr.

    1985-08-21

    At the investigation of Sandia Albuquerque, a semiautomatic powder loader was designed and fabricated for pyrotechnics devices. The basic functions of the system were to load a precise, measured amount of powder into a charge holder and to compact the mixture to a specified density. This report documents the history, rationale, design, and performance of the Mod 1 loader.

  4. Determination of Critical Rock Mass in a Bucket of a Dinting Loader

    NASA Astrophysics Data System (ADS)

    Remiorz, Eryk

    2017-09-01

    The extraction of hard coal deposits lying in increasing depth causes significant problems with maintenance of roadways (maingates, tailgates, etc.). The reduction of the cross section of such excavations, caused by the floor upheaval, leads to the occurrence of many problems with transport and ventilation. Dinting loaders are employed to restore the original size of roadways tightened due to the activity of adverse stresses occurring in the rock mass. These are tracked machines, usually with small width of about 1 m. They often work in roadways with high longitudinal and lateral inclination, as a result of which they are especially susceptible to overturning. The article presents a mathematical model allowing to determine the critical mass of broken rock in a bucket. The model also allows to determine spatial coordinates of a dinting loader's centre of gravity depending on temporary position of movable elements of the loader such as a turntable, boom, coupler and bucket, and depending on the level of loading the bucket with broken rock. It also enables to determine critical angles of the roadways' longitudinal and lateral inclination. The outcomes of computer studies of variations in the position of the loader's centre of gravity depending on deflection angles of moving elements of the loader and the mass of broken rock in the bucket are also presented. Variability ranges of spatial coordinates of the centre of gravity of the loader are also established and examples are given for values of the critical mass of broken rock in the bucket.

  5. Inactivation of the budding yeast cohesin loader Scc2 alters gene expression both globally and in response to a single DNA double strand break

    PubMed Central

    Lindgren, Emma; Hägg, Sara; Giordano, Fosco; Björkegren, Johan; Ström, Lena

    2014-01-01

    Genome integrity is fundamental for cell survival and cell cycle progression. Important mechanisms for keeping the genome intact are proper sister chromatid segregation, correct gene regulation and efficient repair of damaged DNA. Cohesin and its DNA loader, the Scc2/4 complex have been implicated in all these cellular actions. The gene regulation role has been described in several organisms. In yeast it has been suggested that the proteins in the cohesin network would effect transcription based on its role as insulator. More recently, data are emerging indicating direct roles for gene regulation also in yeast. Here we extend these studies by investigating whether the cohesin loader Scc2 is involved in regulation of gene expression. We performed global gene expression profiling in the absence and presence of DNA damage, in wild type and Scc2 deficient G2/M arrested cells, when it is known that Scc2 is important for DNA double strand break repair and formation of damage induced cohesion. We found that not only the DNA damage specific transcriptional response is distorted after inactivation of Scc2 but also the overall transcription profile. Interestingly, these alterations did not correlate with changes in cohesin binding. PMID:25483075

  6. A Transfer of Training Study of Control Loader Dynamics

    NASA Technical Reports Server (NTRS)

    Cardullo, Frank M.; Stanco, Anthony A.; Kelly, Lon C.; Houck, Jacob A.; Grube, Richard C.

    2011-01-01

    The control inceptor used in a simulated vehicle is an important part in maintaining the fidelity of a simulation. The force feedback provided by the control inceptor gives the operator important cues to maintain adequate performance. The dynamics of a control inceptor are typically based on a second order spring mass damper system with damping, force gradient, breakout force, and natural frequency parameters. Changing these parameters can have a great effect on pilot or driver control of the vehicle. The neuromuscular system has a very important role in manipulating the control inceptor within a vehicle. Many studies by McRuer, Aponso, and Hess have dealt with modeling the neuromuscular system and quantifying the effects of a high fidelity control loader as compared to a low fidelity control loader. Humans are adaptive in nature and their control behavior changes based on different control loader dynamics. Humans will change their control behavior to maintain tracking bandwidth and minimize tracking error. This paper reports on a quasi-transfer of training experiment which was performed at the NASA Langley Research Center. The quasi transfer of training study used a high fidelity control loader and a low fidelity control loader. Subjects trained in both simulations and then were transferred to the high fidelity control loader simulation. The parameters for the high fidelity control loader were determined from the literature. The low fidelity control loader parameters were found through testing of a simple computer joystick. A disturbance compensatory task is employed. The compensatory task involves implementing a simple horizon out the window display. A disturbance consisting of a sum of sines is used. The task consists of the subject compensating for the disturbance on the roll angle of the aircraft. The vehicle dynamics are represented as 1/s and 1/s2. The subject will try to maintain level flight throughout the experiment. The subjects consist of non-pilots to remove any effects of pilot experience. First, this paper discusses the implementation of the disturbance compensation task. Second, the high and low fidelity parameters used within the experiment are presented. Finally, an explanation of results from the experiments is presented.

  7. The effect of fragmentation on loader productivity

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

    McGill, M.D.; Freadrich, J.

    1994-12-31

    There are many factors which affect the productivity of a front end loader. One of the most obvious is the characteristics of the material being mined. Characteristics such as size distribution, swell factor or heave, and material hardness/abrasiveness will all effect the efficiency of the mucking cycle. The focus of this paper is to look specifically at the size distribution of blasted material, and quantify the effects on loader productivity.

  8. Comparison of a Skidder and Front-End Loader for Primary Transport of Short-Rotation Trees

    Treesearch

    Raffaele Spinelli; Bruce R. Hartsough

    1999-01-01

    We time-studied a Cat 950F and a Cat 528 grapple skidder as extraction devices for moving bunched whole trees to a landing in a short rotation eucalyptus plantation. The front-end loader was 40 to 60% more productive than the grapple skidder, depending on extraction distance. Alternatively, the single loader could both extract trees and handle the landing duties such...

  9. Assessing musculoskeletal disorders among municipal waste loaders of Mumbai, India.

    PubMed

    Salve, Pradeep; Chokhandre, Praveen; Bansod, Dhananjay

    2017-10-06

    The study aims to assess the impact of municipal waste loading occupation upon developing musculoskeletal disorders (MSDs) and thereby disabilities among waste loaders. Additionally, the study has identified the potential risk factors raising MSDs and disabilities. A cross-sectional case-control design survey was conducted in 6 out of 24 municipal wards of Mumbai during March-September 2015. The study population consisted of municipal waste loaders (N = 180) and a control group (N = 180). The Standardized Modified Nordic questionnaire was adopted to measures the MSDs and thereby disabilities in the past 12 months. A Propensity Score Matching (PSM) method was applied to assess the impact of waste loading occupation on developing MSDs and disabilities. Waste loaders had a significantly higher risk of developing MSDs as well as disabilities than the control group particularly for low back, hip/ thigh upper back and shoulder. Propensity Score Matching results revealed that the MSDs were significantly higher among waste loaders for hip/thigh (22%), low back (19%), shoulder (18%), and upper back (15%) than matched control group. Likewise, MSDs-related disabilities were found to be significantly higher among waste loaders for low back (20%), hip/ thigh (18%) upper back (13%) and shoulder (8%) than the control group. Duration of work, substance use and mental health were found to be the potential psychosocial factors for developing the risk of MSDs and disabilities. The municipal waste loading occupation raised the risk of MSDs and related disabilities among waste loaders compared to the control group. The preventive and curative measures are strongly recommended to minimize the burden of MSDs and disabilities. Int J Occup Med Environ Health 2017;30(6):875-886. This work is available in Open Access model and licensed under a CC BY-NC 3.0 PL license.

  10. Fuelcell-Hybrid Mine loader (LHD)

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

    James L Dippo; Tim Erikson; Kris Hess

    2009-07-10

    The fuel cell hybrid mine loader project, sponsored by a government-industry consortium, was implemented to determine the viability of proton exchange membrane (PEM) fuel cells in underground mining applications. The Department of Energy (DOE) sponsored this project with cost-share support from industry. The project had three main goals: (1) to develop a mine loader powered by a fuel cell, (2) to develop associated metal-hydride storage and refueling systems, and (3) to demonstrate the fuel cell hybrid loader in an underground mine in Nevada. The investigation of a zero-emissions fuel cell power plant, the safe storage of hydrogen, worker health advantagesmore » (over the negative health effects associated with exposure to diesel emissions), and lower operating costs are all key objectives for this project.« less

  11. 30 CFR 56.14101 - Brakes.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... particular piece of equipment. Front-end loaders shall be tested with the loader bucket empty. Equipment... speed the equipment is traveling except for equipment which is designed for the power train to be...

  12. 30 CFR 56.14101 - Brakes.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... particular piece of equipment. Front-end loaders shall be tested with the loader bucket empty. Equipment... speed the equipment is traveling except for equipment which is designed for the power train to be...

  13. 30 CFR 56.14101 - Brakes.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... particular piece of equipment. Front-end loaders shall be tested with the loader bucket empty. Equipment... speed the equipment is traveling except for equipment which is designed for the power train to be...

  14. 30 CFR 56.14101 - Brakes.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... particular piece of equipment. Front-end loaders shall be tested with the loader bucket empty. Equipment... speed the equipment is traveling except for equipment which is designed for the power train to be...

  15. 30 CFR 57.14101 - Brakes.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    .... Front-end loaders shall be tested with the loader bucket empty. Equipment shall not be tested when... equipment which is designed for the power train to be disengaged during braking. (v) Testing speeds shall be...

  16. 30 CFR 57.14101 - Brakes.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    .... Front-end loaders shall be tested with the loader bucket empty. Equipment shall not be tested when... equipment which is designed for the power train to be disengaged during braking. (v) Testing speeds shall be...

  17. 30 CFR 57.14101 - Brakes.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    .... Front-end loaders shall be tested with the loader bucket empty. Equipment shall not be tested when... equipment which is designed for the power train to be disengaged during braking. (v) Testing speeds shall be...

  18. 30 CFR 57.14101 - Brakes.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    .... Front-end loaders shall be tested with the loader bucket empty. Equipment shall not be tested when... equipment which is designed for the power train to be disengaged during braking. (v) Testing speeds shall be...

  19. Molecular mechanism of DNA replication-coupled inactivation of the initiator protein in Escherichia coli: interaction of DnaA with the sliding clamp-loaded DNA and the sliding clamp-Hda complex.

    PubMed

    Su'etsugu, Masayuki; Takata, Makoto; Kubota, Toshio; Matsuda, Yusaku; Katayama, Tsutomu

    2004-06-01

    In Escherichia coli, the ATP-DnaA protein initiates chromosomal replication. After the DNA polymerase III holoenzyme is loaded on to DNA, DnaA-bound ATP is hydrolysed in a manner depending on Hda protein and the DNA-loaded form of the DNA polymerase III sliding clamp subunit, which yields ADP-DnaA, an inactivated form for initiation. This regulatory DnaA-inactivation represses extra initiation events. In this study, in vitro replication intermediates and structured DNA mimicking replicational intermediates were first used to identify structural prerequisites in the process of DnaA-ATP hydrolysis. Unlike duplex DNA loaded with sliding clamps, primer RNA-DNA heteroduplexes loaded with clamps were not associated with DnaA-ATP hydrolysis, and duplex DNA provided in trans did not rescue this defect. At least 40-bp duplex DNA is competent for the DnaA-ATP hydrolysis when a single clamp was loaded. The DnaA-ATP hydrolysis was inhibited when ATP-DnaA was tightly bound to a DnaA box-bearing oligonucleotide. These results imply that the DnaA-ATP hydrolysis involves the direct interaction of ATP-DnaA with duplex DNA flanking the sliding clamp. Furthermore, Hda protein formed a stable complex with the sliding clamp. Based on these, we suggest a mechanical basis in the DnaA-inactivation that ATP-DnaA interacts with the Hda-clamp complex with the aid of DNA binding. Copyright Blackwell Publishing Limited

  20. Automatic Test, Configuration, and Repair of Cellular Arrays

    DTIC Science & Technology

    1975-06-01

    one cell to loed any good cell that is not walled off by flawed cells. The loading Information which the computer sends to the array may select one...to loed that cell’s loader and function-specificatioi state bits. After the cell is loaded» it« loader state bits may specify which of the cell’s...Mechanism With Options FUNCTION-SPEC, STATE BITS D.OUT INO INI OUTO 0UT1 LOO LO PAGE 99 LOADER TIP BITS CHANGER ill LSTA S.U.IN S.R.IN

  1. Selective Arterial Clamping Versus Hilar Clamping for Minimally Invasive Partial Nephrectomy.

    PubMed

    Yezdani, Mona; Yu, Sue-Jean; Lee, David I

    2016-05-01

    Partial nephrectomy has become an accepted treatment of cT1 renal masses as it provides improved long-term renal function compared to radical nephrectomy (Campbell et al. J Urol. 182:1271-9, 2009). Hilar clamping is utilized to help reduce bleeding and improve visibility during tumor resection. However, concern over risk of kidney injury with hilar clamping has led to new techniques to reduce length of warm ischemia time (WIT) during partial nephrectomy. These techniques have progressed over the years starting with early hilar unclamping, controlled hypotension during tumor resection, selective arterial clamping, minimal margin techniques, and off-clamp procedures. Selective arterial clamping has progressed significantly over the years. The main question is what are the exact short- and long-term renal effects from increasing clamp time. Moreover, does it make sense to perform these more time-consuming or more complex procedures if there is no long-term preservation of kidney function? More recent studies have shown no difference in renal function 6 months from surgery when selective arterial clamping or even hilar clamping is employed, although there is short-term improved decline in estimated glomerular filtration rate (eGFR) with selective clamping and off-clamp techniques (Komninos et al. BJU Int. 115:921-8, 2015; Shah et al. 117:293-9, 2015; Kallingal et al. BJU Int. doi: 10.1111/bju.13192, 2015). This paper reviews the progression of total hilar clamping to selective arterial clamping (SAC) and the possible difference its use makes on long-term renal function. SAC may be attempted based on surgeon's decision-making, but may be best used for more complex, larger, more central or hilar tumors and in patients who have renal insufficiency at baseline or a solitary kidney.

  2. 9. PHOTOCOPY, ARCHITECTURAL SECTIONS AND DETAIL DRAWING OF UNDERGROUND STORAGE ...

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

    9. PHOTOCOPY, ARCHITECTURAL SECTIONS AND DETAIL DRAWING OF UNDERGROUND STORAGE MAGAZINES AND LAUNCHER-LOADER ASSEMBLIES. - NIKE Missile Base SL-40, Underground Storage Magazines & Launcher-Loader Assemblies, Southwesternmost end of launch area, Hecker, Monroe County, IL

  3. An abstract class loader for the SSP and its implementation in TL.

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

    Wickstrom, Gregory Lloyd; Winter, Victor Lono; Fraij, Fares

    2004-08-01

    The SSP is a hardware implementation of a subset of the JVM for use in high consequence embedded applications. In this context, a majority of the activities belonging to class loading, as it is defined in the specification of the JVM, can be performed statically. Static class loading has the net result of dramatically simplifying the design of the SSP as well as increasing its performance. Due to the high consequence nature of its applications, strong evidence must be provided that all aspects of the SSP have been implemented correctly. This includes the class loader. This article explores the possibilitymore » of formally verifying a class loader for the SSP implemented in the strategic programming language TL. Specifically, an implementation of the core activities of an abstract class loader is presented and its verification in ACL2 is considered.« less

  4. 7. PHOTOCOPY, PLUMBING AND MECHANICAL PLAN AND DETAILS FOR UNDERGROUND ...

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

    7. PHOTOCOPY, PLUMBING AND MECHANICAL PLAN AND DETAILS FOR UNDERGROUND STORAGE MAGAZINES AND LAUNCHER-LOADER ASSEMBLIES. - NIKE Missile Base SL-40, Underground Storage Magazines & Launcher-Loader Assemblies, Southwesternmost end of launch area, Hecker, Monroe County, IL

  5. 8. PHOTOCOPY, ARCHITECTURAL FLOOR PLAN AND DETAIL DRAWING OF UNDERGROUND ...

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

    8. PHOTOCOPY, ARCHITECTURAL FLOOR PLAN AND DETAIL DRAWING OF UNDERGROUND STORAGE MAGAZINES AND LAUNCHER-LOADER ASSEMBLIES. - NIKE Missile Base SL-40, Underground Storage Magazines & Launcher-Loader Assemblies, Southwesternmost end of launch area, Hecker, Monroe County, IL

  6. 14 CFR 158.3 - Definitions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... passenger by other parties. Ground support equipment means service and maintenance equipment used at an airport to support aeronautical operations and related activities. Baggage tugs, belt loaders, cargo loaders, forklifts, fuel trucks, lavatory trucks, and pushback tractors are among the types of vehicles...

  7. Quality control mechanisms exclude incorrect polymerases from the eukaryotic replication fork

    PubMed Central

    Schauer, Grant D.; O’Donnell, Michael E.

    2017-01-01

    The eukaryotic genome is primarily replicated by two DNA polymerases, Pol ε and Pol δ, that function on the leading and lagging strands, respectively. Previous studies have established recruitment mechanisms whereby Cdc45-Mcm2-7-GINS (CMG) helicase binds Pol ε and tethers it to the leading strand, and PCNA (proliferating cell nuclear antigen) binds tightly to Pol δ and recruits it to the lagging strand. The current report identifies quality control mechanisms that exclude the improper polymerase from a particular strand. We find that the replication factor C (RFC) clamp loader specifically inhibits Pol ε on the lagging strand, and CMG protects Pol ε against RFC inhibition on the leading strand. Previous studies show that Pol δ is slow and distributive with CMG on the leading strand. However, Saccharomyces cerevisiae Pol δ–PCNA is a rapid and processive enzyme, suggesting that CMG may bind and alter Pol δ activity or position it on the lagging strand. Measurements of polymerase binding to CMG demonstrate Pol ε binds CMG with a Kd value of 12 nM, but Pol δ binding CMG is undetectable. Pol δ, like bacterial replicases, undergoes collision release upon completing replication, and we propose Pol δ–PCNA collides with the slower CMG, and in the absence of a stabilizing Pol δ–CMG interaction, the collision release process is triggered, ejecting Pol δ on the leading strand. Hence, by eviction of incorrect polymerases at the fork, the clamp machinery directs quality control on the lagging strand and CMG enforces quality control on the leading strand. PMID:28069954

  8. A Loader for Executing Multi-Binary Applications on the Thinking Machines CM-5: It's Not Just for SPMD Anymore

    NASA Technical Reports Server (NTRS)

    Becker, Jeffrey C.

    1995-01-01

    The Thinking Machines CM-5 platform was designed to run single program, multiple data (SPMD) applications, i.e., to run a single binary across all nodes of a partition, with each node possibly operating on different data. Certain classes of applications, such as multi-disciplinary computational fluid dynamics codes, are facilitated by the ability to have subsets of the partition nodes running different binaries. In order to extend the CM-5 system software to permit such applications, a multi-program loader was developed. This system is based on the dld loader which was originally developed for workstations. This paper provides a high level description of dld, and describes how it was ported to the CM-5 to provide support for multi-binary applications. Finally, it elaborates how the loader has been used to implement the CM-5 version of MPIRUN, a portable facility for running multi-disciplinary/multi-zonal MPI (Message-Passing Interface Standard) codes.

  9. Whole-body vibration in heavy equipment operators of a front-end loader: role of task exposure and tire configuration with and without traction chains.

    PubMed

    Blood, Ryan P; Rynell, Patrik W; Johnson, Peter W

    2012-12-01

    This study measured whole-body vibration (WBV) exposures in front-end loader operators, and evaluated the effects of traction chains and work tasks on their WBV exposures. WBV exposures were measured and compared across three different front-end loader tire configurations: (a) stock rubber tires, (b) rubber tires with ladder chains, and (c) rubber tires with basket chains. The operators completed three distinct standardized tasks: driving on a city street, simulated plowing, and a simulated scooping and dumping task. A portable data acquisition system collected tri-axial time weighted and raw WBV data per ISO 2631-1 and 2631-5 standards. In addition, Global Positioning System (GPS) data were collected in order to compare loader speeds across tire conditions and the standardized tasks. Relative to the stock rubber tires, both types of tire chains significantly increased WBV exposures with the ladder chains having substantially higher WBV exposures compared to basket chains. Additionally, there were task dependent differences in WBV exposures. During the driving task, the z-axis (up and down) was the predominant exposure; the plowing task had a more even distribution of exposure across all three axes; while during scooping and dumping task, the x-axis (fore and aft) had the highest WBV exposures. The GPS data indicated that there were significant speed differences across tasks but not between the basket and ladder chain conditions. Tires with ladder chains increased the front-end loader operators' exposure to WBV above the ISO 2631-1 recommended eight hour action limit increasing risk for adverse health effects. Although more expensive, basket chains are recommended over ladder chains since they substantially lowered the front-end loader operator's exposures and may ultimately reduce vibration related wear and tear on the vehicle. In order to reduce a heavy equipment vehicle (HEV) operator's chances for developing low back pain, this study provides information that health and safety professionals can use to reduce whole-body vibration (WBV) exposures when operating front-end wheel loaders with traction chains. Copyright © 2012 National Safety Council and Elsevier Ltd. All rights reserved.

  10. InterMine Webservices for Phytozome (Rev2)

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

    Carlson, Joseph; Goodstein, David; Rokhsar, Dan

    2014-07-10

    A datawarehousing framework for information provides a useful infrastructure for providers and users of genomic data. For providers, the infrastructure give them a consistent mechanism for extracting raw data. While for the users, the web services supported by the software allows them to make complex, and often unique, queries of the data. Previously, phytozome.net used BioMart to provide the infrastructure. As the complexity, scale and diversity of the dataset as grown, we decided to implement an InterMine web service on our servers. This change was largely motivated by the ability to have a more complex table structure and richer webmore » reporting mechanism than BioMart. For InterMine to achieve its more complex database schema it requires an XML description of the data and an appropriate loader. Unlimited one-to-many and many-to-many relationship between the tables can be enabled in the schema. We have implemented support for:1.) Genomes and annotations for the data in Phytozome. This set is the 48 organisms currently stored in a back end CHADO datastore. The data loaders are modified versions of the CHADO data adapters from FlyMine. 2.) Interproscan results from all proteins in the Phytozome database. 3.) Clusters of proteins into a grouped heirarchically by similarity. 4.) Cufflinks results from tissue-specific RNA-Seq data of Phytozome organisms. 5.) Diversity data (GATK and SnpEFF results) from a set of individual organism. The last two datatypes are new in this implementation of our web services. We anticipate that the scale of these data will increase considerably in the near future.« less

  11. Protein associations in DnaA-ATP hydrolysis mediated by the Hda-replicase clamp complex.

    PubMed

    Su'etsugu, Masayuki; Shimuta, Toh-Ru; Ishida, Takuma; Kawakami, Hironori; Katayama, Tsutomu

    2005-02-25

    In Escherichia coli, the activity of ATP-bound DnaA protein in initiating chromosomal replication is negatively controlled in a replication-coordinated manner. The RIDA (regulatory inactivation of DnaA) system promotes DnaA-ATP hydrolysis to produce the inactivated form DnaA-ADP in a manner depending on the Hda protein and the DNA-loaded form of the beta-sliding clamp, a subunit of the replicase holoenzyme. A highly functional form of Hda was purified and shown to form a homodimer in solution, and two Hda dimers were found to associate with a single clamp molecule. Purified mutant Hda proteins were used in a staged in vitro RIDA system followed by a pull-down assay to show that Hda-clamp binding is a prerequisite for DnaA-ATP hydrolysis and that binding is mediated by an Hda N-terminal motif. Arg(168) in the AAA(+) Box VII motif of Hda plays a role in stable homodimer formation and in DnaA-ATP hydrolysis, but not in clamp binding. Furthermore, the DnaA N-terminal domain is required for the functional interaction of DnaA with the Hda-clamp complex. Single cells contain approximately 50 Hda dimers, consistent with the results of in vitro experiments. These findings and the features of AAA(+) proteins, including DnaA, suggest the following model. DnaA-ATP is hydrolyzed at a binding interface between the AAA(+) domains of DnaA and Hda; the DnaA N-terminal domain supports this interaction; and the interaction of DnaA-ATP with the Hda-clamp complex occurs in a catalytic mode.

  12. 29 CFR 782.5 - Loaders.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... come within the exemption as a partial-duty loader. (Levinson v. Spector Motor Service, 330 U.S. 649... limited handling of them, that his activities will not come within the kind of “loading” which directly... it is light in weight, probably could not be loaded in a manner which would adversely affect “safety...

  13. 29 CFR 782.5 - Loaders.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... come within the exemption as a partial-duty loader. (Levinson v. Spector Motor Service, 330 U.S. 649... limited handling of them, that his activities will not come within the kind of “loading” which directly... it is light in weight, probably could not be loaded in a manner which would adversely affect “safety...

  14. 25. FRONT END LOADERS MOMENTARILY IN REPOSE IN THE ORE ...

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

    25. FRONT END LOADERS MOMENTARILY IN REPOSE IN THE ORE STORAGE YARD. AN ORE BRIDGE THAT FORMERLY TRANSFERRED ORE WITHIN THE STORAGE YARD WAS DESTROYED BY A BLIZZARD IN 1978. - Pennsylvania Railway Ore Dock, Lake Erie at Whiskey Island, approximately 1.5 miles west of Public Square, Cleveland, Cuyahoga County, OH

  15. Safer methods of hauling coalface power loaders

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

    Whitworth, K.

    1976-01-01

    That power loader chains are a menace is evidenced by the Report of H. M. Chief Inspector of Mines and Quarries in Great Britain. In 1974, power loader haulage chains were responsible for 40 percent of all machinery accidents, with one person killed and 18 seriously injured compared with 23 seriously injured in 1973. Half the accidents occurred in the vicinity of the power loader sprockets when persons were struck by the end of a broken chain or were trapped between the chain and the sprocket. The present static haulage chain in tension along the whole length of face ismore » a continuing hazard and the most significant safety contribution that can be made is to design alternative means of haulage to eliminate the chain. Alternative means of haulage have or are being developed and equipment from six machinery manufacturers, from CERCHAR, the French research and development organization, and from the British National Coal Board were demonstrated at the National Coal Board's Swadlincote Test Site. A brief description of each item of equipment is given.« less

  16. Measurement of Spindle Rigidity by using a Magnet Loader

    NASA Astrophysics Data System (ADS)

    Yamazaki, Taku; Matsubara, Atsushi; Fujita, Tomoya; Muraki, Toshiyuki; Asano, Kohei; Kawashima, Kazuyuki

    The static rigidity of a rotating spindle in the radial direction is investigated in this research. A magnetic loading device (magnet loader) has been developed for the measurement. The magnet loader, which has coils and iron cores, generates the electromagnetic force and attracts a dummy tool attached to the spindle. However, the eddy current is generated in the dummy tool with the spindle rotation and reduces the attractive force at high spindle speed. In order to understand the magnetic flux and eddy current in the dummy tool, the electromagnetic field analysis by FEM was carried out. Grooves on the attraction surface of the dummy tool were designed to cut the eddy current flow. The dimension of the groove were decided based on the FEM analysis, and the designed tool were manufactured and tested. The test result shows that the designed tool successfully reduces the eddy current and recovers the attractive force. By using the magnet loader and the grooved tool, the spindle rigidity can be measured when the spindle rotates with a speed up to 10,000 min-1.

  17. Justification of process of loading coal onto face conveyors by auger heads of shearer-loader machines

    NASA Astrophysics Data System (ADS)

    Nguyen, K. L.; Gabov, V. V.; Zadkov, D. A.; Le, T. B.

    2018-03-01

    This paper analyzes the processes of removing coal from the area of its dislodging and loading the disintegrated mass onto face conveyors by auger heads of shearer-loader machines. The loading process is assumed to consist of four subprocesses: dislodging coal, removal of the disintegrated mass by auger blades from the crushing area, passive transportation of the disintegrated mass, and forming the load flow on the bearing surface of a face conveyor. Each of the considered subprocesses is different in its physical nature, the number of factors influencing it, and can be complex or multifactor. Possibilities of improving the efficiency of loading coal onto a face conveyor are addressed. The selected criteria of loading efficiency are load rate, specific energy consumption, and coal size reduction. Efficiency is improved by reducing the resistance to movement of the disintegrated mass during loading by increasing the area of the loading window section and the volume of the loading area on the conveyor, as well as by coordination of intensity of flows related to the considered processes in local areas.

  18. Front End Loader Operator. Open Pit Mining Job Training Series.

    ERIC Educational Resources Information Center

    Savilow, Bill

    This training outline for front end loader operators, one in a series of eight outlines, is designed primarily for company training foremen or supervisors and for trainers to use as an industry-wide guideline for heavy equipment operator training in open pit mining in British Columbia. Intended as a guide for preparation of lesson plans both for…

  19. Comparison of mechanized systems for thinning Ponderosa pine and mixed conifer stands

    Treesearch

    Bruce R. Hartsough; Joseph F. McNeel; Thomas A. Durston; Bryce J. Stokes

    1994-01-01

    We studied three systems for thinning pine plantations and naturally-regenerated stands on the Stanislaus National Forest, California. All three produced small sawlogs and fuel chips. The whole tree system consisted of a feller buncher, skidder, stroke processor, loader and chipper. The cut-to-length system included a harvester, forwarder, loader and chipper. A hybrid...

  20. Comparison of mechanized systems for thinning Ponderosa pine and mixed conifer stands

    Treesearch

    Bruce R. Hartsough; Joseph F. McNeel; Thomas A. Durston; Bryce J. Stokes

    1994-01-01

    Three systems for thinning pine plantations and naturally-regenerated stands were studied. All three produced small sawlogs and fuel chips. The whole-tree system consisted of a feller buncher, skidder, stroke processor, loader, and chipper. The cut-to-length system included a harvester, forwarder, loader, and chipper. A hybrid system combined a feller buncher,...

  1. Command-line cellular electrophysiology for conventional and real-time closed-loop experiments.

    PubMed

    Linaro, Daniele; Couto, João; Giugliano, Michele

    2014-06-15

    Current software tools for electrophysiological experiments are limited in flexibility and rarely offer adequate support for advanced techniques such as dynamic clamp and hybrid experiments, which are therefore limited to laboratories with a significant expertise in neuroinformatics. We have developed lcg, a software suite based on a command-line interface (CLI) that allows performing both standard and advanced electrophysiological experiments. Stimulation protocols for classical voltage and current clamp experiments are defined by a concise and flexible meta description that allows representing complex waveforms as a piece-wise parametric decomposition of elementary sub-waveforms, abstracting the stimulation hardware. To perform complex experiments lcg provides a set of elementary building blocks that can be interconnected to yield a large variety of experimental paradigms. We present various cellular electrophysiological experiments in which lcg has been employed, ranging from the automated application of current clamp protocols for characterizing basic electrophysiological properties of neurons, to dynamic clamp, response clamp, and hybrid experiments. We finally show how the scripting capabilities behind a CLI are suited for integrating experimental trials into complex workflows, where actual experiment, online data analysis and computational modeling seamlessly integrate. We compare lcg with two open source toolboxes, RTXI and RELACS. We believe that lcg will greatly contribute to the standardization and reproducibility of both simple and complex experiments. Additionally, on the long run the increased efficiency due to a CLI will prove a great benefit for the experimental community. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Association between photosynthesis and contrasting features of minor veins in leaves of summer annuals loading phloem via symplastic versus apoplastic routes.

    PubMed

    Muller, Onno; Cohu, Christopher M; Stewart, Jared J; Protheroe, Johanna A; Demmig-Adams, Barbara; Adams, William W

    2014-09-01

    Foliar vascular anatomy and photosynthesis were evaluated for a number of summer annual species that either load sugars into the phloem via a symplastic route (Cucumis sativus L. cv. Straight Eight; Cucurbita pepo L. cv. Italian Zucchini Romanesco; Citrullus lanatus L. cv. Faerie Hybrid; Cucurbita pepo L. cv. Autumn Gold) or an apoplastic route (Nicotiana tabacum L.; Solanum lycopersicum L. cv. Brandywine; Gossypium hirsutum L.; Helianthus annuus L. cv. Soraya), as well as winter annual apoplastic loaders (Spinacia oleracea L. cv. Giant Nobel; Arabidopsis thaliana (L.) Heynhold Col-0, Swedish and Italian ecotypes). For all summer annuals, minor vein cross-sectional xylem area and tracheid number as well as the ratio of phloem loading cells to phloem sieve elements, each when normalized for foliar vein density (VD), was correlated with photosynthesis. These links presumably reflect (1) the xylem's role in providing water to meet foliar transpirational demand supporting photosynthesis and (2) the importance of the driving force of phloem loading as well as the cross-sectional area for phloem sap flux to match foliar photosynthate production. While photosynthesis correlated with the product of VD and cross-sectional phloem cell area among symplastic loaders, photosynthesis correlated with the product of VD and phloem cell number per vein among summer annual apoplastic loaders. Phloem cell size has thus apparently been a target of selection among symplastic loaders (where loading depends on enzyme concentration within loading cells) versus phloem cell number among apoplastic loaders (where loading depends on membrane transporter numbers). © 2014 Scandinavian Plant Physiology Society.

  3. Production and Costs of the Chambers Delimbinator in First Thinning of Pine Plantations

    Treesearch

    Scott T. Mooney; Kevin D. Boston; W. Dale Greene

    2000-01-01

    Production and quality measures were collected and analyzed for a new chain-flail delimbing machine, Chambers Delimbinator, on two operations in central and southwestern Georgia. The first operation, Logger A, used two skidders and one loader while the second operation, Logger B, used two skidder and two loaders. Lobber B with the lower cycle time and larger piece...

  4. Pesticide Safety for Non-Certified Mixers, Loaders and Applicators = Uso Seguro de Pesticidas para Mezcladores, Cargadores y Aplicadores no Certificados.

    ERIC Educational Resources Information Center

    Poli, Bonnie; Fluker, Sam S.

    Written in English and Spanish and completely illustrated, this manual provides basic safety information for pesticide workers. Mixers, loaders, and applicators work with pesticides at their greatest strength and have the highest risk of poisoning. Understanding the pesticide label is the first step to pesticide safety. The words…

  5. Microbiologic contamination during dental radiographic film processing.

    PubMed

    Stanczyk, D A; Paunovich, E D; Broome, J C; Fatone, M A

    1993-07-01

    This study investigated microbiologic contamination of an automatic dental radiograph processor and daylight loader during a week of simulated clinical use. Pure cultures of Candida albicans, Streptococcus pneumoniae, Staphylococcus aureus, or Klebsiella pneumoniae were used to contaminate 320 vinyl intraoral radiograph packets. Each end of the films was deliberately contaminated during opening. These films and 24 uncontaminated control films were processed. Daylight loader ports, inlet and outlet rollers, fixer and developer samples, and 12 processed films were cultured daily. To simulate a weekend, the processor sites were cultured during 72 hours of inactivity after the contaminated runs. The results showed that contamination of the processor and daylight loader occurred and remained even after 48 hours of inactivity. Films remained contaminated after processing. In addition, cross-contamination of films occurred in the processor.

  6. Cutter-loader apparatus having overhung shearer drum

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

    Groger, H.; Harms, E.E.

    1984-05-01

    A longwall mining machine includes a drum cutter-loader and face conveyor wherein the drum cutter-loader is overhung and is supported by a support arm adjacent to the mine face. Nozzles direct high pressure liquid jets against the forward edge of the support arm to cut away the mining face and permit the face side support arm to advance as the mining machine advances. In one embodiment the nozzles are provided along an inclined cutting edge at the forward end of the support arm. Such nozzles may be fixed or oscillating. In an alternative embodiment the nozzles are provided in themore » cylindrical edge zone of the shearer drum and direct the high pressure fluid jets against the cutter edge at the forward end of the support arm.« less

  7. New insights into cohesin loading.

    PubMed

    Litwin, Ireneusz; Wysocki, Robert

    2018-02-01

    Cohesin is a conserved, ring-shaped protein complex that encircles sister chromatids and ensures correct chromosome segregation during mitosis and meiosis. It also plays a crucial role in the regulation of gene expression, DNA condensation, and DNA repair through both non-homologous end joining and homologous recombination. Cohesins are spatiotemporally regulated by the Scc2-Scc4 complex which facilitates cohesin loading onto chromatin at specific chromosomal sites. Over the last few years, much attention has been paid to cohesin and cohesin loader as it became clear that even minor disruptions of these complexes may lead to developmental disorders and cancers. Here we summarize recent developments in the structure of Scc2-Scc4 complex, cohesin loading process, and mediators that determine the Scc2-Scc4 binding patterns to chromatin.

  8. Fleet/TYCOM Level Survey of Armament Handling Problems - Task One of the Naval Aviation Armament Support Equipment Program Management Study

    DTIC Science & Technology

    1977-06-21

    7. AUTHOR(#) 6. CONTRACT OR GRANT NUMBER(#) PILA /UDERIAN 9. PERFORMING ORGANIZATION NAME AND ADDRESS 10. PROGRAM ELEMENT. PROJECT, TASK...mechanically complex (i.e. contains an internal combustion engine) and/or comes in direct support of the aircraft is managed and maintained by the AIMD. The...an internal combustion engine. Accordingly, only the Aero 33D/E Trailer, Aero 51B Trailer, 21A/C Bomb Skid, and Aero 47A Weapons Loader are maintained

  9. TypeLoader: A fast and efficient automated workflow for the annotation and submission of novel full-length HLA alleles.

    PubMed

    Surendranath, V; Albrecht, V; Hayhurst, J D; Schöne, B; Robinson, J; Marsh, S G E; Schmidt, A H; Lange, V

    2017-07-01

    Recent years have seen a rapid increase in the discovery of novel allelic variants of the human leukocyte antigen (HLA) genes. Commonly, only the exons encoding the peptide binding domains of novel HLA alleles are submitted. As a result, the IPD-IMGT/HLA Database lacks sequence information outside those regions for the majority of known alleles. This has implications for the application of the new sequencing technologies, which deliver sequence data often covering the complete gene. As these technologies simplify the characterization of the complete gene regions, it is desirable for novel alleles to be submitted as full-length sequences to the database. However, the manual annotation of full-length alleles and the generation of specific formats required by the sequence repositories is prone to error and time consuming. We have developed TypeLoader to address both these facets. With only the full-length sequence as a starting point, Typeloader performs automatic sequence annotation and subsequently handles all steps involved in preparing the specific formats for submission with very little manual intervention. TypeLoader is routinely used at the DKMS Life Science Lab and has aided in the successful submission of more than 900 novel HLA alleles as full-length sequences to the European Nucleotide Archive repository and the IPD-IMGT/HLA Database with a 95% reduction in the time spent on annotation and submission when compared with handling these processes manually. TypeLoader is implemented as a web application and can be easily installed and used on a standalone Linux desktop system or within a Linux client/server architecture. TypeLoader is downloadable from http://www.github.com/DKMS-LSL/typeloader. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  10. Mission-Based Analyses of Armor Training Requirements. Volume 7. Training Objectives for the XM1 Loader

    DTIC Science & Technology

    1982-04-01

    the gas particulate filter system MODULE L: OPERATE THE M250 BRENADE LAUNCHER 1L. Load the grenade launcher 2L. Unload the grenade launcher MODULE M...k Initia~ng Stimulus: Thei (11rdLr from the T.C. to load the M250 .p grenade launcher. J ACTION Loader will: 1L. Load the grenade launcher. 2L. Unload

  11. Handler, bystander and reentry exposure to TCDD from application of Agent Orange by C-123 aircraft during the Vietnam War.

    PubMed

    Ross, John H; Hewitt, Andrew; Armitage, James; Solomon, Keith; Watkins, Deborah K; Ginevan, Michael E

    2015-02-01

    Using validated models and methods routinely employed by pesticide regulatory agencies, the absorbed dosages of Agent Orange (AO) herbicide contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were estimated for mixer/loaders, applicators, and individuals in the vicinity of applications of AO by C-123 aircraft during the Vietnam War. Resulting dosages of TCDD were then transformed to estimates of adipose residues, and compared to population biomonitoring of known mixer/loaders and applicators as well as ground troops in Vietnam and civilians in the U.S. Results demonstrate that mixer/loaders and applicators had the greatest exposures and their measured residues of TCDD in adipose were consistent with the estimated exposures. Further, the potentially exposed ground troops, including those who could have been directly sprayed during aerial defoliation, had measured adipose residues that were consistent with those in civilian U.S. populations with no defined source of exposure exposures and both of those cohorts had orders of magnitude less exposure than the mixer/loaders or applicators. Despite the availability of validated exposure modeling methods for decades, the quantitative TCDD dose estimates presented here are the first of their kind for the Vietnam conflict. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. GFU-6/E 30mm Ammunition Loader for GAU-8A Gun System

    DTIC Science & Technology

    1975-12-01

    dropped onto the conveyor (See Figure 17) and transferred to the aircraft on these elements through flexible chuting which interfaces directly with the...14. Dwell Linkage 19 15. Horizontal Conveyor - Flex Chute Interface 20 16. Element Turnaround Sprocket 21 17. Horizontal Conveyor 22 18. Load Head... conveyor element mechanism for transport to the GAU-8/A loader unit. d. The contractor shall design the automated loading system to accept unfired

  13. Modernization of Deployable Airfield Debris Removal Equipment

    DTIC Science & Technology

    2017-04-01

    cleared the debris away from the crater by lowering the front bucket and setting the bottom blade at a 45-deg angle. The debris was pushed...the Komatsu WA 150 baseline loader. The crater clearing process involved the loader orienting the bucket blade at approximately 45 deg to the...wider than the track’s width (Figure 30). This caused debris to run under the track and lift the equipment, potentially throwing the operator. The final

  14. The National Shipbuilding Research Program: Solid Waste Segregation and Recycling

    DTIC Science & Technology

    1998-03-01

    Shields , Recycling Coordinator D.C. Department of Public Works 65 K Street, NE Washington, DC 20002 202-727-5887 Task Three, Tab Three Page 42 George...SHEAR X FRONT END LOADERS = CONVEYORS X FORKLIFTS O WEIGHT SCALES X PROCESSING DROP-BALL BREAKAGE X CUTTING TORCHES GAS = PLASMA = POWDER = WATER-JET...Loaders Conveyors Forklifts Weight Scales Processing Drop-ball Breakage Cutting Torches Gas Plasma Powder Laser Water-jet Abrasive disk Shears Ferrous

  15. Improved memory loading techniques for the TSRV display system

    NASA Technical Reports Server (NTRS)

    Easley, W. C.; Lynn, W. A.; Mcluer, D. G.

    1986-01-01

    A recent upgrade of the TSRV research flight system at NASA Langley Research Center retained the original monochrome display system. However, the display memory loading equipment was replaced requiring design and development of new methods of performing this task. This paper describes the new techniques developed to load memory in the display system. An outdated paper tape method for loading the BOOTSTRAP control program was replaced by EPROM storage of the characters contained on the tape. Rather than move a tape past an optical reader, a counter was implemented which steps sequentially through EPROM addresses and presents the same data to the loader circuitry. A cumbersome cassette tape method for loading the applications software was replaced with a floppy disk method using a microprocessor terminal installed as part of the upgrade. The cassette memory image was transferred to disk and a specific software loader was written for the terminal which duplicates the function of the cassette loader.

  16. Improvement of loading and transport bodies of tunneling machines

    NASA Astrophysics Data System (ADS)

    Nosenko, A. S.; Isakov, V. S.; Domnitskiy, A. A.; Shemshura, E. A.; Zubov, V. V.

    2017-10-01

    The article discusses the use of sets of equipment for mining and construction of road tunnels with the use of a self-propelled vehicle and bunker loaders to transport the mined rock. A patented design of the tunneling combine loading body, which allows adjusting its performance and power consumption in the loading mode, is considered. Reducing stress in extreme situations will make it possible to avoid overloading the drive and increase the reliability of the combine as a whole. The scheme of one of the most promising variants of the wedge bunker loader with variable geometry of the conveying elements is presented. The proposed design solves the problem of “locking” the material in narrow parts of the conveying body or in places where loading and transport bodies of different width are connected, as well as in the blast-and-heap mode of the loader.

  17. Wrapping SRS with CORBA: from textual data to distributed objects.

    PubMed

    Coupaye, T

    1999-04-01

    Biological data come in very different shapes. Databanks are maintained and used by distinct organizations. Text is the de facto Standard exchange format. The SRS system can integrate heterogeneous textual databanks but it was lacking a way to structure the extracted data. This paper presents a CORBA interface to the SRS system which manages databanks in a flat file format. SRS Object Servers are CORBA wrappers for SRS. They allow client applications (visualisation tools, data mining tools, etc.) to access and query SRS servers remotely through an Object Request Broker (ORB). They provide loader objects that contain the information extracted from the databanks by SRS. Loader objects are not hard-coded but generated in a flexible way by using loader specifications which allow SRS administrators to package data coming from distinct databanks. The prototype may be available for beta-testing. Please contact the SRS group (http://srs.ebi.ac.uk).

  18. Adult Competency Education Kit. Basic Skills in Speaking, Math, and Reading for Employment. Part R. ACE Competency Based Job Descriptions: #95--Bus Driver; #98--General Loader; #99--Forklift Operator; #100--Material Handler.

    ERIC Educational Resources Information Center

    San Mateo County Office of Education, Redwood City, CA. Career Preparation Centers.

    This fifteenth of fifteen sets of Adult Competency Education (ACE) Competency Based Job Descriptions in the ACE kit contains job descriptions for Bus Driver, General Loader, Forklift Operator, and Material Handler. Each begins with a fact sheet that includes this information: occupational title, D.O.T. code, ACE number, career ladder, D.O.T.…

  19. Elan4/SPARC V9 Cross Loader and Dynamic Linker

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

    anf Fabien Lebaillif-Delamare, Fabrizio Petrini

    2004-10-25

    The Elan4/Sparc V9 Croos Loader and Liner is a part of the Linux system software that allows the dynamic loading and linking of user code in the network interface Quadrics QsNETII, also called as Elan4 Quadrics. Elan44 uses a thread processor that is based on the assembly instruction set of the Sparc V9. All this software is integrated as a Linux kernel module in the Linux 2.6.5 release.

  20. Evaluation of Wheel Loaders in Open Pit Marble Quarrying by Using the AHP and Topsis Approaches / Ocena pracy ładowarki na podwoziu kołowym w odkrywkowej kopalni marmuru w oparciu o metody AHP i topsis

    NASA Astrophysics Data System (ADS)

    Kun, Mete; Topaloǧlu, Şeyda; Malli, Tahir

    2013-03-01

    The marble mining in Turkey has been rising since the early 80's. In relation to that, the marble income has become noticeably bigger than those of other mining sectors. In recent years, marble and natural stone export composes half of the total mine export with a value of two billion dollars. This rapid development observed in marble operation has increased the importance of mining economics, income-expenditure balance and cost analysis. The most important cost elements observed in marble quarrying are machinery and equipment, labor costs and geological structures of the field. The aim of this study is to is to propose a multi-criteria decision making (MCDM) approach to evaluate the wheel loader alternatives and select the best loader under multiple criteria. A two-step methodology based on two MCDM methods, which are namely the Analytic Hierarchy Process (AHP) and the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS), are used in the evaluation procedure. More precisely, AHP is applied to determine the relative weights of evaluation criteria and TOPSIS is applied to rank the wheel loader alternatives. The proposed approach also provides a relatively simple and very well suited decision making tool for this type of decision making problems.

  1. Structural Basis of Transcription Inhibition by Fidaxomicin (Lipiarmycin A3).

    PubMed

    Lin, Wei; Das, Kalyan; Degen, David; Mazumder, Abhishek; Duchi, Diego; Wang, Dongye; Ebright, Yon W; Ebright, Richard Y; Sineva, Elena; Gigliotti, Matthew; Srivastava, Aashish; Mandal, Sukhendu; Jiang, Yi; Liu, Yu; Yin, Ruiheng; Zhang, Zhening; Eng, Edward T; Thomas, Dennis; Donadio, Stefano; Zhang, Haibo; Zhang, Changsheng; Kapanidis, Achillefs N; Ebright, Richard H

    2018-04-05

    Fidaxomicin is an antibacterial drug in clinical use for treatment of Clostridium difficile diarrhea. The active ingredient of fidaxomicin, lipiarmycin A3 (Lpm), functions by inhibiting bacterial RNA polymerase (RNAP). Here we report a cryo-EM structure of Mycobacterium tuberculosis RNAP holoenzyme in complex with Lpm at 3.5-Å resolution. The structure shows that Lpm binds at the base of the RNAP "clamp." The structure exhibits an open conformation of the RNAP clamp, suggesting that Lpm traps an open-clamp state. Single-molecule fluorescence resonance energy transfer experiments confirm that Lpm traps an open-clamp state and define effects of Lpm on clamp dynamics. We suggest that Lpm inhibits transcription by trapping an open-clamp state, preventing simultaneous interaction with promoter -10 and -35 elements. The results account for the absence of cross-resistance between Lpm and other RNAP inhibitors, account for structure-activity relationships of Lpm derivatives, and enable structure-based design of improved Lpm derivatives. Copyright © 2018 Elsevier Inc. All rights reserved.

  2. Logic Design of a Shared Disk System in a Multi-Micro Computer Environment.

    DTIC Science & Technology

    1983-06-01

    overall system, is given. An exnaustive description of eacn device can De found in tne cited references. A. INTEL 80S5 Tne INTEL Be86 is a nign...eitner could De accomplished, it was necessary to understand ootn tne existing system arcnitecture ani software. Tne last cnapter addressed tnat...to De adapted: tne loader program and tne Doot ROP program. Tne loader program is a simplified version of CP/M-Bö and contains cniy encu^n file

  3. Avoiding the Health Hazard of People from Construction Vehicles: A Strategy for Controlling the Vibration of a Wheel Loader

    PubMed Central

    Chi, Feng; Zhou, Jun; Zhang, Qi; Wang, Yong; Huang, Panling

    2017-01-01

    The vibration control of a construction vehicle must be carried out in order to meet the aims of sustainable environmental development and to avoid the potential human health hazards. In this paper, based on market feedback, the driver seat vibration of a type of wheel loader in the left and right direction, is found to be significant over a certain speed range. In order to find abnormal vibration components, the order tracking technique (OTT) and transmission path analysis (TPA) were used to analyze the vibration sources of the wheel loader. Through this analysis, it can be seen that the abnormal vibration comes from the interaction between the tire tread and the road, and this is because the vibration was amplified by the cab mount, which was eventually transmitted to the cab seat. Finally, the seat vibration amplitudes were decreased by up to 50.8%, after implementing the vibration reduction strategy. PMID:28282849

  4. Avoiding the Health Hazard of People from Construction Vehicles: A Strategy for Controlling the Vibration of a Wheel Loader.

    PubMed

    Chi, Feng; Zhou, Jun; Zhang, Qi; Wang, Yong; Huang, Panling

    2017-03-08

    The vibration control of a construction vehicle must be carried out in order to meet the aims of sustainable environmental development and to avoid the potential human health hazards. In this paper, based on market feedback, the driver seat vibration of a type of wheel loader in the left and right direction, is found to be significant over a certain speed range. In order to find abnormal vibration components, the order tracking technique (OTT) and transmission path analysis (TPA) were used to analyze the vibration sources of the wheel loader. Through this analysis, it can be seen that the abnormal vibration comes from the interaction between the tire tread and the road, and this is because the vibration was amplified by the cab mount, which was eventually transmitted to the cab seat. Finally, the seat vibration amplitudes were decreased by up to 50.8%, after implementing the vibration reduction strategy.

  5. Force-controlled patch clamp of beating cardiac cells.

    PubMed

    Ossola, Dario; Amarouch, Mohamed-Yassine; Behr, Pascal; Vörös, János; Abriel, Hugues; Zambelli, Tomaso

    2015-03-11

    From its invention in the 1970s, the patch clamp technique is the gold standard in electrophysiology research and drug screening because it is the only tool enabling accurate investigation of voltage-gated ion channels, which are responsible for action potentials. Because of its key role in drug screening, innovation efforts are being made to reduce its complexity toward more automated systems. While some of these new approaches are being adopted in pharmaceutical companies, conventional patch-clamp remains unmatched in fundamental research due to its versatility. Here, we merged the patch clamp and atomic force microscope (AFM) techniques, thus equipping the patch-clamp with the sensitive AFM force control. This was possible using the FluidFM, a force-controlled nanopipette based on microchanneled AFM cantilevers. First, the compatibility of the system with patch-clamp electronics and its ability to record the activity of voltage-gated ion channels in whole-cell configuration was demonstrated with sodium (NaV1.5) channels. Second, we showed the feasibility of simultaneous recording of membrane current and force development during contraction of isolated cardiomyocytes. Force feedback allowed for a gentle and stable contact between AFM tip and cell membrane enabling serial patch clamping and injection without apparent cell damage.

  6. Mispair-specific Recruitment of the Mlh1-Pms1 Complex Identifies Repair Substrates of the Saccharomyces cerevisiae Msh2-Msh3 Complex*

    PubMed Central

    Srivatsan, Anjana; Bowen, Nikki; Kolodner, Richard D.

    2014-01-01

    DNA mismatch repair is initiated by either the Msh2-Msh6 or the Msh2-Msh3 mispair recognition heterodimer. Here we optimized the expression and purification of Saccharomyces cerevisiae Msh2-Msh3 and performed a comparative study of Msh2-Msh3 and Msh2-Msh6 for mispair binding, sliding clamp formation, and Mlh1-Pms1 recruitment. Msh2-Msh3 formed sliding clamps and recruited Mlh1-Pms1 on +1, +2, +3, and +4 insertion/deletions and CC, AA, and possibly GG mispairs, whereas Msh2-Msh6 formed mispair-dependent sliding clamps and recruited Mlh1-Pms1 on 7 of the 8 possible base:base mispairs, the +1 insertion/deletion mispair, and to a low level on the +2 but not the +3 or +4 insertion/deletion mispairs and not on the CC mispair. The mispair specificity of sliding clamp formation and Mlh1-Pms1 recruitment but not mispair binding alone correlated best with genetic data on the mispair specificity of Msh2-Msh3- and Msh2-Msh6-dependent mismatch repair in vivo. Analysis of an Msh2-Msh6/Msh3 chimeric protein and mutant Msh2-Msh3 complexes showed that the nucleotide binding domain and communicating regions but not the mispair binding domain of Msh2-Msh3 are responsible for the extremely rapid dissociation of Msh2-Msh3 sliding clamps from DNA relative to that seen for Msh2-Msh6, and that amino acid residues predicted to stabilize Msh2-Msh3 interactions with bent, strand-separated mispair-containing DNA are more critical for the recognition of small +1 insertion/deletions than larger +4 insertion/deletions. PMID:24550389

  7. Mispair-specific recruitment of the Mlh1-Pms1 complex identifies repair substrates of the Saccharomyces cerevisiae Msh2-Msh3 complex.

    PubMed

    Srivatsan, Anjana; Bowen, Nikki; Kolodner, Richard D

    2014-03-28

    DNA mismatch repair is initiated by either the Msh2-Msh6 or the Msh2-Msh3 mispair recognition heterodimer. Here we optimized the expression and purification of Saccharomyces cerevisiae Msh2-Msh3 and performed a comparative study of Msh2-Msh3 and Msh2-Msh6 for mispair binding, sliding clamp formation, and Mlh1-Pms1 recruitment. Msh2-Msh3 formed sliding clamps and recruited Mlh1-Pms1 on +1, +2, +3, and +4 insertion/deletions and CC, AA, and possibly GG mispairs, whereas Msh2-Msh6 formed mispair-dependent sliding clamps and recruited Mlh1-Pms1 on 7 of the 8 possible base:base mispairs, the +1 insertion/deletion mispair, and to a low level on the +2 but not the +3 or +4 insertion/deletion mispairs and not on the CC mispair. The mispair specificity of sliding clamp formation and Mlh1-Pms1 recruitment but not mispair binding alone correlated best with genetic data on the mispair specificity of Msh2-Msh3- and Msh2-Msh6-dependent mismatch repair in vivo. Analysis of an Msh2-Msh6/Msh3 chimeric protein and mutant Msh2-Msh3 complexes showed that the nucleotide binding domain and communicating regions but not the mispair binding domain of Msh2-Msh3 are responsible for the extremely rapid dissociation of Msh2-Msh3 sliding clamps from DNA relative to that seen for Msh2-Msh6, and that amino acid residues predicted to stabilize Msh2-Msh3 interactions with bent, strand-separated mispair-containing DNA are more critical for the recognition of small +1 insertion/deletions than larger +4 insertion/deletions.

  8. Distortion and Residual Stress Control in Integrally Stiffened Structure Produced by Direct Metal Deposition

    NASA Technical Reports Server (NTRS)

    Lin, Shih-Yung; Hoffman, Eric K.; Domack, Marcia S.

    2007-01-01

    2-D thermo-mechanical model developed to characterize distortion and residual stresses in integral structure produced by DMD. Demonstrated as a tool to guide experimental development of DMD fabrication process for aero structures. Distortion and residual stresses are local to deposit. Most distortion develops during deposition of the first few layers; Little change in distortion or residual stresses after fifth deposit layer Most of distortion is localized just beneath the build. Thicker build plates and the use of build lands results in greatest decrease in levels of distortion. Pre-straining shown to reduce distortion. Difficult to implement, particularly for complex stiffener arrays. Clamp position has complex effect on distortion and stresses. Overall distortion reduced with decreasing clamp clearance. Larger clamp clearances induce bending. Use of pre-heat and active cooling show minor influence on panel distortion. Generate changes in thermal gradients in the build plate.

  9. Use of Semi-Autonomous Tools for ISS Commanding and Monitoring

    NASA Technical Reports Server (NTRS)

    Brzezinski, Amy S.

    2014-01-01

    As the International Space Station (ISS) has moved into a utilization phase, operations have shifted to become more ground-based with fewer mission control personnel monitoring and commanding multiple ISS systems. This shift to fewer people monitoring more systems has prompted use of semi-autonomous console tools in the ISS Mission Control Center (MCC) to help flight controllers command and monitor the ISS. These console tools perform routine operational procedures while keeping the human operator "in the loop" to monitor and intervene when off-nominal events arise. Two such tools, the Pre-positioned Load (PPL) Loader and Automatic Operators Recorder Manager (AutoORM), are used by the ISS Communications RF Onboard Networks Utilization Specialist (CRONUS) flight control position. CRONUS is responsible for simultaneously commanding and monitoring the ISS Command & Data Handling (C&DH) and Communications and Tracking (C&T) systems. PPL Loader is used to uplink small pieces of frequently changed software data tables, called PPLs, to ISS computers to support different ISS operations. In order to uplink a PPL, a data load command must be built that contains multiple user-input fields. Next, a multiple step commanding and verification procedure must be performed to enable an onboard computer for software uplink, uplink the PPL, verify the PPL has incorporated correctly, and disable the computer for software uplink. PPL Loader provides different levels of automation in both building and uplinking these commands. In its manual mode, PPL Loader automatically builds the PPL data load commands but allows the flight controller to verify and save the commands for future uplink. In its auto mode, PPL Loader automatically builds the PPL data load commands for flight controller verification, but automatically performs the PPL uplink procedure by sending commands and performing verification checks while notifying CRONUS of procedure step completion. If an off-nominal condition occurs during procedure execution, PPL Loader notifies CRONUS through popup messages, allowing CRONUS to examine the situation and choose an option of how PPL loader should proceed with the procedure. The use of PPL Loader to perform frequent, routine PPL uplinks offloads CRONUS to better monitor two ISS systems. It also reduces procedure performance time and decreases risk of command errors. AutoORM identifies ISS communication outage periods and builds commands to lock, playback, and unlock ISS Operations Recorder files. Operation Recorder files are circular buffer files of continually recorded ISS telemetry data. Sections of these files can be locked from further writing, be played back to capture telemetry data that occurred during an ISS loss of signal (LOS) period, and then be unlocked for future recording use. Downlinked Operation Recorder files are used by mission support teams for data analysis, especially if failures occur during LOS. The commands to lock, playback, and unlock Operations Recorder files are encompassed in three different operational procedures and contain multiple user-input fields. AutoORM provides different levels of automation for building and uplinking the commands to lock, playback, and unlock Operations Recorder files. In its automatic mode, AutoORM automatically detects ISS LOS periods, then generates and uplinks the commands to lock, playback, and unlock Operations Recorder files when MCC regains signal with ISS. AutoORM also features semi-autonomous and manual modes which integrate CRONUS more into the command verification and uplink process. AutoORMs ability to automatically detect ISS LOS periods and build the necessary commands to preserve, playback, and release recorded telemetry data greatly offloads CRONUS to perform more high-level cognitive tasks, such as mission planning and anomaly troubleshooting. Additionally, since Operations Recorder commands contain numerical time input fields which are tedious for a human to manually build, AutoORM's ability to automatically build commands reduces operational command errors. PPL Loader and AutoORM demonstrate principles of semi-autonomous operational tools that will benefit future space mission operations. Both tools employ different levels of automation to perform simple and routine procedures, thereby offloading human operators to perform higher-level cognitive tasks. Because both tools provide procedure execution status and highlight off-nominal indications, the flight controller is able to intervene during procedure execution if needed. Semi-autonomous tools and systems that can perform routine procedures, yet keep human operators informed of execution, will be essential in future long-duration missions where the onboard crew will be solely responsible for spacecraft monitoring and control.

  10. Exposure to airborne organophosphates originating from hydraulic and turbine oils among aviation technicians and loaders.

    PubMed

    Solbu, Kasper; Daae, Hanne Line; Thorud, Syvert; Ellingsen, Dag Gunnar; Lundanes, Elsa; Molander, Paal

    2010-12-01

    This study describes the potential for occupational exposure to organophosphates (OPs) originating from turbine and hydraulic oils, among ground personnel within the aviation industry. The OPs tri-n-butyl phosphate (TnBP), dibutyl phenyl phosphate (DBPP), triphenyl phosphate (TPP) and tricresyl phosphate (TCP) have been emphasized due to their use in such oils. Oil aerosol/vapor and total volatile organic compounds (tVOCs) in air were also determined. In total, 228 and 182 OPs and oil aerosol/vapor samples from technician and loader work tasks during work on 42 and 21 aircrafts, respectively, were collected in pairs. In general, the measured exposure levels were below the limit of quantification (LOQ) for 84%/98% (oil aerosol) and 82%/90% (TCP) of the samples collected during technician/loader work tasks. The air concentration ranges for all samples related to technician work were

  11. Remote drill bit loader

    DOEpatents

    Dokos, J.A.

    1997-12-30

    A drill bit loader is described for loading a tapered shank of a drill bit into a similarly tapered recess in the end of a drill spindle. The spindle has a transverse slot at the inner end of the recess. The end of the tapered shank of the drill bit has a transverse tang adapted to engage in the slot so that the drill bit will be rotated by the spindle. The loader is in the form of a cylinder adapted to receive the drill bit with the shank projecting out of the outer end of the cylinder. Retainer pins prevent rotation of the drill bit in the cylinder. The spindle is lowered to extend the shank of the drill bit into the recess in the spindle and the spindle is rotated to align the slot in the spindle with the tang on the shank. A spring unit in the cylinder is compressed by the drill bit during its entry into the recess of the spindle and resiliently drives the tang into the slot in the spindle when the tang and slot are aligned. 5 figs.

  12. Remote drill bit loader

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

    Dokos, James A.

    A drill bit loader for loading a tapered shank of a drill bit into a similarly tapered recess in the end of a drill spindle. The spindle has a transverse slot at the inner end of the recess. The end of the tapered shank of the drill bit has a transverse tang adapted to engage in the slot so that the drill bit will be rotated by the spindle. The loader is in the form of a cylinder adapted to receive the drill bit with the shank projecting out of the outer end of the cylinder. Retainer pins prevent rotationmore » of the drill bit in the cylinder. The spindle is lowered to extend the shank of the drill bit into the recess in the spindle and the spindle is rotated to align the slot in the spindle with the tang on the shank. A spring unit in the cylinder is compressed by the drill bit during its entry into the recess of the spindle and resiliently drives the tang into the slot in the spindle when the tang and slot are aligned.« less

  13. Remote drill bit loader

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

    Dokos, J.A.

    A drill bit loader is described for loading a tapered shank of a drill bit into a similarly tapered recess in the end of a drill spindle. The spindle has a transverse slot at the inner end of the recess. The end of the tapered shank of the drill bit has a transverse tang adapted to engage in the slot so that the drill bit will be rotated by the spindle. The loader is in the form of a cylinder adapted to receive the drill bit with the shank projecting out of the outer end of the cylinder. Retainer pinsmore » prevent rotation of the drill bit in the cylinder. The spindle is lowered to extend the shank of the drill bit into the recess in the spindle and the spindle is rotated to align the slot in the spindle with the tang on the shank. A spring unit in the cylinder is compressed by the drill bit during its entry into the recess of the spindle and resiliently drives the tang into the slot in the spindle when the tang and slot are aligned. 5 figs.« less

  14. Prototyping Faithful Execution in a Java virtual machine.

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

    Tarman, Thomas David; Campbell, Philip LaRoche; Pierson, Lyndon George

    2003-09-01

    This report presents the implementation of a stateless scheme for Faithful Execution, the design for which is presented in a companion report, ''Principles of Faithful Execution in the Implementation of Trusted Objects'' (SAND 2003-2328). We added a simple cryptographic capability to an already simplified class loader and its associated Java Virtual Machine (JVM) to provide a byte-level implementation of Faithful Execution. The extended class loader and JVM we refer to collectively as the Sandia Faithfully Executing Java architecture (or JavaFE for short). This prototype is intended to enable exploration of more sophisticated techniques which we intend to implement in hardware.

  15. Evaluation of Grounding Impedance of a Complex Lightning Protective System Using Earth Ground Clamp Measurements and ATP Modeling

    NASA Technical Reports Server (NTRS)

    Mata, Carlos T.; Rakov, V. A.; Mata, Angel G.

    2010-01-01

    A new Lightning Protection System (LPS) was designed and built at Launch Complex 39B (LC39B), at the Kennedy Space Center (KSC), Florida, which consists of a catenary wire system (at a height of about 181 meters above ground level) supported by three insulators installed atop three towers in a triangular configuration. A total of nine downconductors (each about 250 meters long, on average) are connected to the catenary wire system. Each of the nine downconductors is connected to a 7.62-meter radius circular counterpoise conductor with six equally spaced 6-meter long vertical grounding rods. Grounding requirements at LC39B call for all underground and above ground metallic piping, enclosures, raceways, and cable trays, within 7.62 meters of the counterpoise, to be bounded to the counterpoise, which results in a complex interconnected grounding system, given the many metallic piping, raceways, and cable trays that run in multiple direction around LC39B. The complexity of this grounding system makes the fall of potential method, which uses multiple metallic rods or stakes, unsuitable for measuring the grounding impedances of the downconductors. To calculate the downconductors grounding impedance, an Earth Ground Clamp (a stakeless grounding resistance measuring device) and a LPS Alternative Transient Program (ATP) model are used. The Earth Ground Clamp is used to measure the loop impedance plus the grounding impedance of each downconductor and the ATP model is used to calculate the loop impedance of each downconductor circuit. The grounding impedance of the downconductors is then calculated by subtracting the ATP calculated loop impedances from the Earth Ground Clamp measurements.

  16. Establishment method of a mixture model and its practical application for transmission gears in an engineering vehicle

    NASA Astrophysics Data System (ADS)

    Wang, Jixin; Wang, Zhenyu; Yu, Xiangjun; Yao, Mingyao; Yao, Zongwei; Zhang, Erping

    2012-09-01

    Highly versatile machines, such as wheel loaders, forklifts, and mining haulers, are subject to many kinds of working conditions, as well as indefinite factors that lead to the complexity of the load. The load probability distribution function (PDF) of transmission gears has many distributions centers; thus, its PDF cannot be well represented by just a single-peak function. For the purpose of representing the distribution characteristics of the complicated phenomenon accurately, this paper proposes a novel method to establish a mixture model. Based on linear regression models and correlation coefficients, the proposed method can be used to automatically select the best-fitting function in the mixture model. Coefficient of determination, the mean square error, and the maximum deviation are chosen and then used as judging criteria to describe the fitting precision between the theoretical distribution and the corresponding histogram of the available load data. The applicability of this modeling method is illustrated by the field testing data of a wheel loader. Meanwhile, the load spectra based on the mixture model are compiled. The comparison results show that the mixture model is more suitable for the description of the load-distribution characteristics. The proposed research improves the flexibility and intelligence of modeling, reduces the statistical error and enhances the fitting accuracy, and the load spectra complied by this method can better reflect the actual load characteristic of the gear component.

  17. An analysis of injuries to front-end loader operators during ingress and egress.

    PubMed

    Nasarwanji, Mahiyar F; Pollard, Jonisha; Porter, William

    2018-05-01

    Slips, trips, and falls from mobile mining equipment have been documented for decades. However, little research has been conducted to determine the events precipitating these incidents during ingress or egress. This study examined slips, trips, and falls sustained during ingress or egress from front-end loaders to determine the frequencies of factors that may contribute to injuries. Non-fatal injuries, when getting on or off of front-end wheel loaders specifically, were identified, coded, and analyzed from the Mine Safety and Health Administration's accidents, injuries, and illnesses database. Overall trends, events that precipitated the injury, injuries sustained, contributing factors, location of the individual, and equipment characteristics were analyzed. More incidents occurred during egress (63%); and egress is believed to be more hazardous than ingress. Foot slips were the most common event that precipitated the incident and the leading cause of these was contaminants on the equipment. Misstep, loss of footing, and step on/in related incidents were more common during egress and are likely due to the operator's reduced visibility when descending a ladder facing the equipment, limiting their ability to detect hazards. Egress also makes an operator less capable of avoiding unsafe ground conditions as indicated by the significant number of step on/in injuries occurring on the ground during egress. Most of the front-end loaders associated with the incidents were found to have bottom rungs with flexible rails, which may also increase fall risk during egress due to inconsistent rung heights and lengthy transition areas from the ground, through the flexible-railed rungs, to the rungs with rigid rails. Recommendations are provided to reduce the risk for slips, trips, and falls from mobile mining equipment.

  18. Testing of New Materials and Computer Aided Optimization of Process Parameters and Clamping Device During Predevelopment of Laser Welding Processes

    NASA Astrophysics Data System (ADS)

    Weidinger, Peter; Günther, Kay; Fitzel, Martin; Logvinov, Ruslan; Ilin, Alexander; Ploshikhin, Vasily; Hugger, Florian; Mann, Vincent; Roth, Stephan; Schmidt, Michael

    The necessity for weight reduction in motor vehicles in order to save fuel consumption pushes automotive suppliers to use materials of higher strength. Due to their excellent crash behavior high strength steels are increasingly applied in various structures. In this paper some predevelopment steps for a material change from a micro alloyed to dual phase and complex phase steels of a T-joint assembly are displayed. Initially the general weldability of the materials regarding pore formation, hardening in the heat affected zone and hot cracking susceptibility is discussed. After this basic investigation, the computer aided design optimization of a clamping device is shown, in which influences of the clamping jaw, the welding position and the clamping forces upon weld quality are presented. Finally experimental results of the welding process are displayed, which validate the numerical simulation.

  19. Direct Observation of Markovian Behavior of the Mechanical Unfolding of Individual Proteins

    PubMed Central

    Cao, Yi; Kuske, Rachel; Li, Hongbin

    2008-01-01

    Single-molecule force-clamp spectroscopy is a valuable tool to analyze unfolding kinetics of proteins. Previous force-clamp spectroscopy experiments have demonstrated that the mechanical unfolding of ubiquitin deviates from the generally assumed Markovian behavior and involves the features of glassy dynamics. Here we use single molecule force-clamp spectroscopy to study the unfolding kinetics of a computationally designed fast-folding mutant of the small protein GB1, which shares a similar β-grasp fold as ubiquitin. By treating the mechanical unfolding of polyproteins as the superposition of multiple identical Poisson processes, we developed a simple stochastic analysis approach to analyze the dwell time distribution of individual unfolding events in polyprotein unfolding trajectories. Our results unambiguously demonstrate that the mechanical unfolding of NuG2 fulfills all criteria of a memoryless Markovian process. This result, in contrast with the complex mechanical unfolding behaviors observed for ubiquitin, serves as a direct experimental demonstration of the Markovian behavior for the mechanical unfolding of a protein and reveals the complexity of the unfolding dynamics among structurally similar proteins. Furthermore, we extended our method into a robust and efficient pseudo-dwell-time analysis method, which allows one to make full use of all the unfolding events obtained in force-clamp experiments without categorizing the unfolding events. This method enabled us to measure the key parameters characterizing the mechanical unfolding energy landscape of NuG2 with improved precision. We anticipate that the methods demonstrated here will find broad applications in single-molecule force-clamp spectroscopy studies for a wide range of proteins. PMID:18375518

  20. Modelling and study of active vibration control for off-road vehicle

    NASA Astrophysics Data System (ADS)

    Zhang, Junwei; Chen, Sizhong

    2014-05-01

    In view of special working characteristics and structure, engineering machineries do not have conventional suspension system typically. Consequently, operators have to endure severe vibrations which are detrimental both to their health and to the productivity of the loader. Based on displacement control, a kind of active damping method is developed for a skid-steer loader. In this paper, the whole hydraulic system for active damping method is modelled which include swash plate dynamics model, proportional valve model, piston accumulator model, pilot-operated check valve model, relief valve model, pump loss model, and cylinder model. A new road excitation model is developed for the skid-steer loader specially. The response of chassis vibration acceleration to road excitation is verified through simulation. The simulation result of passive accumulator damping is compared with measurements and the comparison shows that they are close. Based on this, parallel PID controller and track PID controller with acceleration feedback are brought into the simulation model, and the simulation results are compared with passive accumulator damping. It shows that the active damping methods with PID controllers are better in reducing chassis vibration acceleration and pitch movement. In the end, the test work for active damping method is proposed for the future work.

  1. Crystal structure of the DNA polymerase III β subunit (β-clamp) from the extremophile Deinococcus radiodurans.

    PubMed

    Niiranen, Laila; Lian, Kjersti; Johnson, Kenneth A; Moe, Elin

    2015-02-27

    Deinococcus radiodurans is an extremely radiation and desiccation resistant bacterium which can tolerate radiation doses up to 5,000 Grays without losing viability. We are studying the role of DNA repair and replication proteins for this unusual phenotype by a structural biology approach. The DNA polymerase III β subunit (β-clamp) acts as a sliding clamp on DNA, promoting the binding and processivity of many DNA-acting proteins, and here we report the crystal structure of D. radiodurans β-clamp (Drβ-clamp) at 2.0 Å resolution. The sequence verification process revealed that at the time of the study the gene encoding Drβ-clamp was wrongly annotated in the genome database, encoding a protein of 393 instead of 362 amino acids. The short protein was successfully expressed, purified and used for crystallisation purposes in complex with Cy5-labeled DNA. The structure, which was obtained from blue crystals, shows a typical ring-shaped bacterial β-clamp formed of two monomers, each with three domains of identical topology, but with no visible DNA in electron density. A visualisation of the electrostatic surface potential reveals a highly negatively charged outer surface while the inner surface and the dimer forming interface have a more even charge distribution. The structure of Drβ-clamp was determined to 2.0 Å resolution and shows an evenly distributed electrostatic surface charge on the DNA interacting side. We hypothesise that this charge distribution may facilitate efficient movement on encircled DNA and help ensure efficient DNA metabolism in D. radiodurans upon exposure to high doses of ionizing irradiation or desiccation.

  2. Screening of E. coli β-clamp Inhibitors Revealed that Few Inhibit Helicobacter pylori More Effectively: Structural and Functional Characterization.

    PubMed

    Pandey, Preeti; Verma, Vijay; Dhar, Suman Kumar; Gourinath, Samudrala

    2018-01-11

    The characteristic of interaction with various enzymes and processivity-promoting nature during DNA replication makes β-clamp an important drug target. Helicobacter pylori ( H. pylori ) have several unique features in DNA replication machinery that makes it different from other microorganisms. To find out whether difference in DNA replication proteins behavior accounts for any difference in drug response when compared to E. coli , in the present study, we have tested E. coli β-clamp inhibitor molecules against H. pylori β-clamp. Various approaches were used to test the binding of inhibitors to H. pylori β-clamp including docking, surface competition assay, complex structure determination, as well as antimicrobial assay. Out of five shortlisted inhibitor molecules on the basis of docking score, three molecules, 5-chloroisatin, carprofen, and 3,4-difluorobenzamide were co-crystallized with H. pylori β-clamp and the structures show that they bind at the protein-protein interaction site as expected. In vivo studies showed only two molecules, 5-chloroisatin, and 3,4-difluorobenzamide inhibited the growth of the pylori with MIC values in micro molar range, which is better than the inhibitory effect of the same drugs on E. coli . Therefore, the evaluation of such drugs against H. pylori may explore the possibility to use to generate species-specific pharmacophore for development of new drugs against H. pylori .

  3. Unusual planned complex suicide committed with a muzzle-loading pistol in combination with subsequent hanging.

    PubMed

    Ondruschka, Benjamin; Morgenthal, Sylvia; Dreβler, Jan; Bayer, Ronny

    2016-11-01

    In Germany, suicides by firearms are not very common in contrast to deaths by hanging and intoxications. The use of historical muzzle-loading firearms in the context of suicides is a rarity. Contact shots from muzzle loaders cause an unusual wound morphology with extensive soot soiling. We report the case of a 59-year-old man, who committed a planned complex suicide by shooting into his mouth with a replica percussion gun in combination with hanging. The gunshot injury showed strong explosive effects in the oral cavity with fractures of the facial bones and the skull associated with cerebral evisceration (so-called Krönlein shot). Due to the special constellation of the case with hanging immediately after the shot, external bleeding from the head injuries was only moderate. Therefore, the head injuries could be assessed and partially reconstructed already at the scene.

  4. Application of Quality Management Tools for Evaluating the Failure Frequency of Cutter-Loader and Plough Mining Systems

    NASA Astrophysics Data System (ADS)

    Biały, Witold

    2017-06-01

    Failure frequency in the mining process, with a focus on the mining machine, has been presented and illustrated by the example of two coal-mines. Two mining systems have been subjected to analysis: a cutter-loader and a plough system. In order to reduce costs generated by failures, maintenance teams should regularly make sure that the machines are used and operated in a rational and effective way. Such activities will allow downtimes to be reduced, and, in consequence, will increase the effectiveness of a mining plant. The evaluation of mining machines' failure frequency contained in this study has been based on one of the traditional quality management tools - the Pareto chart.

  5. US Army Armor Reference Data in Three Volumes. Volume II. Nondivisional Organizations.

    DTIC Science & Technology

    1981-01-01

    SQuat 1r) R 2 SGI IB20 (Team Ldr) R I SP4 19GIO (Loader/Gnr) P I SP5 19G20 (St Gunner) P 1 t,5 19G20 (Sr Gunner) P I SP4 IICII ()Carrrr Dv) P 2 SP4 11810...Sergeant 2 SP4 Mat Storage Sp 1 SIC Platoon SOT1 SPC Food Svc SGT 1 SP4 Supply Con & Acct Cik 1 SP4 Lt Veh DriverI SFC Motor Sergeant I SP4 Subsistence...PLATOON Ho-.-- Loader Scoop Type 2-1/2 CU YD .Platoon Leader Tank Fabric Fuel 10,000-Gal. 1 SPC Platoon SC? 1 PFlC Reports Clerk 0lRadio Set N/VRC-46

  6. Combined acute hyperglycemic and hyperinsulinemic clamp induced profibrotic and proinflammatory responses in the kidney

    PubMed Central

    DeSilva, Kristin; Sorice, Gian Pio; Muscogiuri, Giovanna; Jimenez, Fabio; Ahuja, Seema; Barnes, Jefferey L.; Choudhury, Goutam Ghosh; Musi, Nicolas; DeFronzo, Ralph; Kasinath, Balakuntalam S.

    2013-01-01

    Increase in matrix protein content in the kidney is a cardinal feature of diabetic kidney disease. While renal matrix protein content is increased by chronic hyperglycemia, whether it is regulated by acute elevation of glucose and insulin has not been addressed. In this study, we aimed to evaluate whether short duration of combined hyperglycemia and hyperinsulinemia, mimicking the metabolic environment of prediabetes and early type 2 diabetes, induces kidney injury. Normal rats were subjected to either saline infusion (control, n = 4) or 7 h of combined hyperglycemic-hyperinsulinemic clamp (HG+HI clamp; n = 6). During the clamp, plasma glucose and plasma insulin were maintained at about 350 mg/dl and 16 ng/ml, respectively. HG+HI clamp increased the expression of renal cortical transforming growth factor-β (TGF-β) and renal matrix proteins, laminin and fibronectin. This was associated with the activation of SMAD3, Akt, mammalian target of rapamycin (mTOR) complexes, and ERK signaling pathways and their downstream target events in the initiation and elongation phases of mRNA translation, an important step in protein synthesis. Additionally, HG+HI clamp provoked renal inflammation as shown by the activation of Toll-like receptor 4 (TLR4) and infiltration of CD68-positive monocytes. Urinary F2t isoprostane excretion, an index of renal oxidant stress, was increased in the HG+HI clamp rats. We conclude that even a short duration of hyperglycemia and hyperinsulinemia contributes to activation of pathways that regulate matrix protein synthesis, inflammation, and oxidative stress in the kidney. This finding could have implications for the control of short-term rises in blood glucose in diabetic individuals at risk of developing kidney disease. PMID:24108867

  7. Combined acute hyperglycemic and hyperinsulinemic clamp induced profibrotic and proinflammatory responses in the kidney.

    PubMed

    Mariappan, Meenalakshmi M; DeSilva, Kristin; Sorice, Gian Pio; Muscogiuri, Giovanna; Jimenez, Fabio; Ahuja, Seema; Barnes, Jefferey L; Choudhury, Goutam Ghosh; Musi, Nicolas; DeFronzo, Ralph; Kasinath, Balakuntalam S

    2014-02-01

    Increase in matrix protein content in the kidney is a cardinal feature of diabetic kidney disease. While renal matrix protein content is increased by chronic hyperglycemia, whether it is regulated by acute elevation of glucose and insulin has not been addressed. In this study, we aimed to evaluate whether short duration of combined hyperglycemia and hyperinsulinemia, mimicking the metabolic environment of prediabetes and early type 2 diabetes, induces kidney injury. Normal rats were subjected to either saline infusion (control, n = 4) or 7 h of combined hyperglycemic-hyperinsulinemic clamp (HG+HI clamp; n = 6). During the clamp, plasma glucose and plasma insulin were maintained at about 350 mg/dl and 16 ng/ml, respectively. HG+HI clamp increased the expression of renal cortical transforming growth factor-β (TGF-β) and renal matrix proteins, laminin and fibronectin. This was associated with the activation of SMAD3, Akt, mammalian target of rapamycin (mTOR) complexes, and ERK signaling pathways and their downstream target events in the initiation and elongation phases of mRNA translation, an important step in protein synthesis. Additionally, HG+HI clamp provoked renal inflammation as shown by the activation of Toll-like receptor 4 (TLR4) and infiltration of CD68-positive monocytes. Urinary F2t isoprostane excretion, an index of renal oxidant stress, was increased in the HG+HI clamp rats. We conclude that even a short duration of hyperglycemia and hyperinsulinemia contributes to activation of pathways that regulate matrix protein synthesis, inflammation, and oxidative stress in the kidney. This finding could have implications for the control of short-term rises in blood glucose in diabetic individuals at risk of developing kidney disease.

  8. π-Clamp-mediated cysteine conjugation

    NASA Astrophysics Data System (ADS)

    Zhang, Chi; Welborn, Matthew; Zhu, Tianyu; Yang, Nicole J.; Santos, Michael S.; van Voorhis, Troy; Pentelute, Bradley L.

    2016-02-01

    Site-selective functionalization of complex molecules is one of the most significant challenges in chemistry. Typically, protecting groups or catalysts must be used to enable the selective modification of one site among many that are similarly reactive, and general strategies that selectively tune the local chemical environment around a target site are rare. Here, we show a four-amino-acid sequence (Phe-Cys-Pro-Phe), which we call the ‘π-clamp’, that tunes the reactivity of its cysteine thiol for site-selective conjugation with perfluoroaromatic reagents. We use the π-clamp to selectively modify one cysteine site in proteins containing multiple endogenous cysteine residues. These examples include antibodies and cysteine-based enzymes that would be difficult to modify selectively using standard cysteine-based methods. Antibodies modified using the π-clamp retained binding affinity to their targets, enabling the synthesis of site-specific antibody-drug conjugates for selective killing of HER2-positive breast cancer cells. The π-clamp is an unexpected approach to mediate site-selective chemistry and provides new avenues to modify biomolecules for research and therapeutics.

  9. Temperature-Controlled Clamping and Releasing Mechanism

    NASA Technical Reports Server (NTRS)

    Rosing, David; Ford, Virginia

    2005-01-01

    A report describes the development of a mechanism that automatically clamps upon warming and releases upon cooling between temperature limits of approx. =180 K and approx. =293 K. The mechanism satisfied a need specific to a program that involved repeated excursions of a spectrometer between a room-temperature atmospheric environment and a cryogenic vacuum testing environment. The mechanism was also to be utilized in the intended application of the spectrometer, in which the spectrometer would be clamped for protection during launch of a spacecraft and released in the cold of outer space to allow it to assume its nominal configuration for scientific observations. The mechanism is passive in the sense that its operation does not depend on a control system and does not require any power other than that incidental to heating and cooling. The clamping and releasing action is effected by bolt-preloaded stacks of shape-memory-alloy (SMA) cylinders. In designing this mechanism, as in designing other, similar SMA mechanisms, it was necessary to account for the complex interplay among thermal expansion, elastic and inelastic deformation under load, and SMA thermomechanical properties.

  10. Invariants of electromechanical coupling coefficients in piezoceramics.

    PubMed

    Mezheritsky, Alex V

    2003-12-01

    The relationships between coefficients of electromechanical coupling (CEMC) of various types of piezoceramic resonator (PR) vibrations are considered. Being constant for a given piezoceramic state, the range of variation of piezoceramics dielectric permittivity from a mechanically "free" condition at relatively low frequencies up to an "overall clamped" condition at high frequencies is determined by a consecutive "clamping", caused by a complex of CEMCs of various particular vibrational modes peculiar to the resonator. As the difference between "free" and "overall clamped" permittivities is always determined by the maximal piezomaterial ki3 coupling coefficient, the difference does not depend on the path that was gone through the low-high frequency range, which includes all the vibrational modes possible for a particular PR. The influence of the piezoelectric and elastic anisotropy of lead-zirconate-titanate (PZT) piezoceramic materials on relative CEMC variations was experimentally investigated.

  11. In Vivo Quantification of Cell Coupling in Plants with Different Phloem-Loading Strategies[W][OA

    PubMed Central

    Liesche, Johannes; Schulz, Alexander

    2012-01-01

    Uptake of photoassimilates into the leaf phloem is the key step in carbon partitioning and phloem transport. Symplasmic and apoplasmic loading strategies have been defined in different plant taxa based on the abundance of plasmodesmata between mesophyll and phloem. For apoplasmic loading to occur, an absence of plasmodesmata is a sufficient but not a necessary criterion, as passage of molecules through plasmodesmata might well be blocked or restricted. Here, we present a noninvasive, whole-plant approach to test symplasmic coupling and quantify the intercellular flux of small molecules using photoactivation microscopy. Quantification of coupling between all cells along the prephloem pathways of the apoplasmic loader Vicia faba and Nicotiana tabacum showed, to our knowledge for the first time in vivo, that small solutes like sucrose can diffuse through plasmodesmata up to the phloem sieve element companion cell complex (SECCC). As expected, the SECCC was found to be symplasmically isolated for small solutes. In contrast, the prephloem pathway of the symplasmic loader Cucurbita maxima was found to be well coupled with the SECCC. Phloem loading in gymnosperms is not well understood, due to a profoundly different leaf anatomy and a scarcity of molecular data compared with angiosperms. A cell-coupling analysis for Pinus sylvestris showed high symplasmic coupling along the entire prephloem pathway, comprising at least seven cell border interfaces between mesophyll and sieve elements. Cell coupling together with measurements of leaf sap osmolality indicate a passive symplasmic loading type. Similarities and differences of this loading type with that of angiosperm trees are discussed. PMID:22422939

  12. 30 CFR 77.1109 - Quantity and location of firefighting equipment.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... construction. (c)(1) Mobile equipment, including trucks, front-end loaders, bulldozers, portable welding units...) Fire extinguishers shall be provided at permanent electrical installations commensurate with the...

  13. Osmotic mechanism of the loop extrusion process

    NASA Astrophysics Data System (ADS)

    Yamamoto, Tetsuya; Schiessel, Helmut

    2017-09-01

    The loop extrusion theory assumes that protein factors, such as cohesin rings, act as molecular motors that extrude chromatin loops. However, recent single molecule experiments have shown that cohesin does not show motor activity. To predict the physical mechanism involved in loop extrusion, we here theoretically analyze the dynamics of cohesin rings on a loop, where a cohesin loader is in the middle and unloaders at the ends. Cohesin monomers bind to the loader rather frequently and cohesin dimers bind to this site only occasionally. Our theory predicts that a cohesin dimer extrudes loops by the osmotic pressure of cohesin monomers on the chromatin fiber between the two connected rings. With this mechanism, the frequency of the interactions between chromatin segments depends on the loading and unloading rates of dimers at the corresponding sites.

  14. Role of the α clamp in the protein translocation mechanism of anthrax toxin

    PubMed Central

    Brown, Michael J.; Thoren, Katie L.; Krantz, Bryan A.

    2015-01-01

    Membrane-embedded molecular machines are utilized to move water-soluble proteins across these barriers. Anthrax toxin forms one such machine through the self-assembly of its three component proteins—protective antigen (PA), lethal factor (LF), and edema factor (EF). Upon endocytosis into host cells, acidification of the endosome induces PA to form a membrane-inserted channel, which unfolds LF and EF and translocates them into the host cytosol. Translocation is driven by the proton motive force, comprised of the chemical potential, the proton-gradient (ΔpH), and the membrane potential (ΔΨ). A crystal structure of the lethal toxin core complex revealed an “α clamp” structure that binds to substrate helices nonspecifically. Here we test the hypothesis that through the recognition of unfolding helical structure the α clamp can accelerate the rate of translocation. We produced a synthetic PA mutant in which an α helix was crosslinked into the α clamp to block its function. This synthetic construct impairs translocation by raising a yet uncharacterized translocation barrier shown to be much less force dependent than the known unfolding barrier. We also report that the α clamp more stably binds substrates that can form helices than those, such as polyproline, that cannot. Hence the α clamp recognizes substrates by a general shape-complementarity mechanism. Substrates that are incapable of forming compact secondary structure (due to the introduction of a polyproline track) are severely deficient for translocation. Therefore, the α clamp and its recognition of helical structure in the translocating substrate play key roles in the molecular mechanism of protein translocation. PMID:26344833

  15. Dendrimer-assisted patch-clamp sizing of nuclear pores

    PubMed Central

    Bustamante, J.O.; Michelette, E.R.F.; Geibel, J.P.; Hanover, J.A.; McDonnell, T.J.; Dean, D.A.

    2015-01-01

    Macromolecular translocation (MMT) across the nuclear envelope (NE) occurs exclusively through the nuclear pore complex (NPC). Therefore, the diameter of the NPC aqueous/electrolytic channel (NPCC) is important for cellular structure and function. The NPCC diameter was previously determined to be ≅10 nm with electron microscopy (EM) using the translocation of colloidal gold particles. Here we present patch-clamp and fluorescence microscopy data from adult cardiomyocyte nuclei that demonstrate the use of patch-clamp for assessing NPCC diameter. Fluorescence microscopy with B-phycoerythrin (BPE, 240 kDa) conjugated to a nuclear localization signal (NLS) demonstrated that these nuclei were competent for NPC-mediated MMT (NPC-MMT). Furthermore, when exposed to an appropriate cell lysate, the nuclei expressed enhanced green fluorescence protein (EGFP) after 5–10 h of incubation with the plasmid for this protein (pEGFP, 3.1 MDa). Nucleus-attached patch-clamp showed that colloidal gold particles were not useful probes; they modified NPCC gating. As a result of this finding, we searched for an inert class of particles that could be used without irreversibly affecting NPCC gating and found that fluorescently labeled Star-burst dendrimers, a distinct class of polymers, were useful. Our patch-clamp and fluorescence microscopy data with calibrated dendrimers indicate that the cardiomyocyte NPCC diameter varies between 8 and 9 nm. These studies open a new direction in the investigation of live, continuous NPC dynamics under physiological conditions. PMID:10784359

  16. Nocturnal Boundary-Layer Phenomena Observed at a Complex Site During the Perdigão Experiment

    NASA Astrophysics Data System (ADS)

    Bell, T.; Klein, P. M.; Smith, E.; Gebauer, J.; Turner, D. D.

    2017-12-01

    The Perdigão Field Experiment set out to study atmospheric flows in complex terrain and to collect a high-quality dataset for the validation of meso- and micro-scale models. An Intensive Observation Period (IOP) was conducted from May 1, 2017 through June 15, 2017 where a multitude of instruments were deployed in and around two nearly parallel ridges. The Collaborative Lower Atmospheric Mobile Profiling System (CLAMPS) was deployed and operated in the valley between the ridges. The CLAMPS facility, which was developed as a joint effort between the School of Meteorology at OU and NOAA's National Severe Storms Laboratory (NSSL), takes advantage of a microwave radiometer (MWR), an atmospheric emitted radiance interferometer (AERI), and a scanning doppler Lidar to profile the boundary layer with a high temporal and spatial resolution. Optimized Lidar scanning strategies and joint retrievals for the MWR and ARI data provide detailed information about the wind, turbulence and thermodynamic structure from the surface up to 1000 m AGL on most nights; sometimes the max height is even higher. Over the course of the IOP, CLAMPS observed many different phenomena. During some nights, when stronger background prevailed and was directed perpendicular to the valley, waves were observed at the ridges and in the valley. At the same time, radiational cooling led to drainage flows in the valley, particularly during nights when the mesoscale forcing was weak. At first, CLAMPS profile observations and data collected with radiosondes released at a near-by site are compared to assess the data quality. The radiosonde observations are also being used to document and classify the upper-level flow during the IOP. Additionally, CLAMPS data from a few selected nights will be presented and analyzed in terms of turbulence and its impact on mixing inside and above the valley. June 1-2 represents a good base-state case. Winds at ridge height were generally less than 5ms-1 after 0Z and valley flows were observed by CLAMPS. On May 15-16, a narrow 10ms-1 jet was present near ridge height and a wave formed in the valley overnight. On May 21-22, another 10ms-1 jet was observed, though flow in the valley was very different. Finally, the impacts of the different flow phenomena on the turbulence structure and atmospheric stability throughout the night will be discussed.

  17. Design and evaluation of a slave manipulator with roll-pitch-roll wrist and automatic tool loading mechanism in telerobotic surgery.

    PubMed

    Kim, Ki-Young; Lee, Jung-Ju

    2012-12-01

    As there is a shortage of scrub nurses in many hospitals, automatic surgical tool exchanging mechanism without human labour has been studied. Minimally invasive robotic surgeries (MIRS) also require scrub nurses. A surgical tool loading mechanism without a scrub nurse's assistance for MIRS is proposed. Many researchers have developed minimally invasive surgical instruments with a wrist joint that can be movable inside the abdomen. However, implementation of a distal rolling joint on a gripper is rare. To implement surgical tool exchanging without a scrub nurse's assistance, a slave manipulator and a tool loader were developed to load and unload a surgical tool unit. A surgical tool unit with a roll-pitch-roll wrist was developed. Several experiments were performed to validate the effectiveness of the slave manipulator and the surgical tool unit. The slave manipulator and the tool loader were able to successfully unload and load the surgical tool unit without human assistance. The total duration of unloading and loading the surgical tool unit was 97 s. Motion tracking experiments of the distal rolling joint were performed. The maximum positioning error of the step input response was 2°. The advantage of the proposed slave manipulator and tool loader is that other robotic systems or human labour are not needed for surgical tool loading. The feasibility of the distal rolling joint in MIS is verified. Copyright © 2012 John Wiley & Sons, Ltd.

  18. Effects of Imperfect Dynamic Clamp: Computational and Experimental Results

    PubMed Central

    Bettencourt, Jonathan C.; Lillis, Kyle P.; White, John A.

    2008-01-01

    In the dynamic clamp technique, a typically nonlinear feedback system delivers electrical current to an excitable cell that represents the actions of “virtual” ion channels (e.g., channels that are gated by local membrane potential or by electrical activity in neighboring biological or virtual neurons). Since the conception of this technique, there have been a number of different implementations of dynamic clamp systems, each with differing levels of flexibility and performance. Embedded hardware-based systems typically offer feedback that is very fast and precisely timed, but these systems are often expensive and sometimes inflexible. PC-based systems, on the other hand, allow the user to write software that defines an arbitrarily complex feedback system, but real-time performance in PC-based systems can be deteriorated by imperfect real-time performance. Here we systematically evaluate the performance requirements for artificial dynamic clamp knock-in of transient sodium and delayed rectifier potassium conductances. Specifically we examine the effects of controller time step duration, differential equation integration method, jitter (variability in time step), and latency (the time lag from reading inputs to updating outputs). Each of these control system flaws is artificially introduced in both simulated and real dynamic clamp experiments. We demonstrate that each of these errors affect dynamic clamp accuracy in a way that depends on the time constants and stiffness of the differential equations being solved. In simulations, time steps above 0.2 ms lead to catastrophic alteration of spike shape, but the frequency-vs.-current relationship is much more robust. Latency (the part of the time step that occurs between measuring membrane potential and injecting re-calculated membrane current) is a crucial factor as well. Experimental data are substantially more sensitive to inaccuracies than simulated data. PMID:18076999

  19. Hda Monomerization by ADP Binding Promotes Replicase Clamp-mediated DnaA-ATP Hydrolysis*S⃞

    PubMed Central

    Su'etsugu, Masayuki; Nakamura, Kenta; Keyamura, Kenji; Kudo, Yuka; Katayama, Tsutomu

    2008-01-01

    ATP-DnaA is the initiator of chromosomal replication in Escherichia coli, and the activity of DnaA is regulated by the regulatory inactivation of the DnaA (RIDA) system. In this system, the Hda protein promotes DnaA-ATP hydrolysis to produce inactive ADP-DnaA in a mechanism that is mediated by the DNA-loaded form of the replicase sliding clamp. In this study, we first revealed that hda translation uses an unusual initiation codon, CUG, located downstream of the annotated initiation codon. The CUG initiation codon could be used for restricting the Hda level, as this initiation codon has a low translation efficiency, and the cellular Hda level is only ∼100 molecules per cell. Hda translated using the correct reading frame was purified and found to have a high RIDA activity in vitro. Moreover, we found that Hda has a high affinity for ADP but not for other nucleotides, including ATP. ADP-Hda was active in the RIDA system in vitro and stable in a monomeric state, whereas apo-Hda formed inactive homomultimers. Both ADP-Hda and apo-Hda could form complexes with the DNA-loaded clamp; however, only ADP-Hda-DNA-clamp complexes were highly functional in the following interaction with DnaA. Formation of ADP-Hda was also observed in vivo, and mutant analysis suggested that ADP binding is crucial for cellular Hda activity. Thus, we propose that ADP is a crucial Hda ligand that promotes the activated conformation of the protein. ADP-dependent monomerization might enable the arginine finger of the Hda AAA+ domain to be accessible to ATP bound to the DnaA AAA+ domain. PMID:18977760

  20. Hda monomerization by ADP binding promotes replicase clamp-mediated DnaA-ATP hydrolysis.

    PubMed

    Su'etsugu, Masayuki; Nakamura, Kenta; Keyamura, Kenji; Kudo, Yuka; Katayama, Tsutomu

    2008-12-26

    ATP-DnaA is the initiator of chromosomal replication in Escherichia coli, and the activity of DnaA is regulated by the regulatory inactivation of the DnaA (RIDA) system. In this system, the Hda protein promotes DnaA-ATP hydrolysis to produce inactive ADP-DnaA in a mechanism that is mediated by the DNA-loaded form of the replicase sliding clamp. In this study, we first revealed that hda translation uses an unusual initiation codon, CUG, located downstream of the annotated initiation codon. The CUG initiation codon could be used for restricting the Hda level, as this initiation codon has a low translation efficiency, and the cellular Hda level is only approximately 100 molecules per cell. Hda translated using the correct reading frame was purified and found to have a high RIDA activity in vitro. Moreover, we found that Hda has a high affinity for ADP but not for other nucleotides, including ATP. ADP-Hda was active in the RIDA system in vitro and stable in a monomeric state, whereas apo-Hda formed inactive homomultimers. Both ADP-Hda and apo-Hda could form complexes with the DNA-loaded clamp; however, only ADP-Hda-DNA-clamp complexes were highly functional in the following interaction with DnaA. Formation of ADP-Hda was also observed in vivo, and mutant analysis suggested that ADP binding is crucial for cellular Hda activity. Thus, we propose that ADP is a crucial Hda ligand that promotes the activated conformation of the protein. ADP-dependent monomerization might enable the arginine finger of the Hda AAA+ domain to be accessible to ATP bound to the DnaA AAA+ domain.

  1. Rv0004 is a new essential member of the mycobacterial DNA replication machinery

    PubMed Central

    Hooppaw, Anna J.; Richardson, Kirill; Lee, Hark Joon; Kimmey, Jacqueline M.; Aldridge, Bree B.

    2017-01-01

    DNA replication is fundamental for life, yet a detailed understanding of bacterial DNA replication is limited outside the organisms Escherichia coli and Bacillus subtilis. Many bacteria, including mycobacteria, encode no identified homologs of helicase loaders or regulators of the initiator protein DnaA, despite these factors being essential for DNA replication in E. coli and B. subtilis. In this study we discover that a previously uncharacterized protein, Rv0004, from the human pathogen Mycobacterium tuberculosis is essential for bacterial viability and that depletion of Rv0004 leads to a block in cell cycle progression. Using a combination of genetic and biochemical approaches, we found that Rv0004 has a role in DNA replication, interacts with DNA and the replicative helicase DnaB, and affects DnaB-DnaA complex formation. We also identify a conserved domain in Rv0004 that is predicted to structurally resemble the N-terminal protein-protein interaction domain of DnaA. Mutation of a single conserved tryptophan within Rv0004’s DnaA N-terminal-like domain leads to phenotypes similar to those observed upon Rv0004 depletion and can affect the association of Rv0004 with DnaB. In addition, using live cell imaging during depletion of Rv0004, we have uncovered a previously unappreciated role for DNA replication in coordinating mycobacterial cell division and cell size. Together, our data support that Rv0004 encodes a homolog of the recently identified DciA family of proteins found in most bacteria that lack the DnaC-DnaI helicase loaders in E. coli and B. subtilis. Therefore, the mechanisms of Rv0004 elucidated here likely apply to other DciA homologs and reveal insight into the diversity of bacterial strategies in even the most conserved biological processes. PMID:29176877

  2. BioWarehouse: a bioinformatics database warehouse toolkit

    PubMed Central

    Lee, Thomas J; Pouliot, Yannick; Wagner, Valerie; Gupta, Priyanka; Stringer-Calvert, David WJ; Tenenbaum, Jessica D; Karp, Peter D

    2006-01-01

    Background This article addresses the problem of interoperation of heterogeneous bioinformatics databases. Results We introduce BioWarehouse, an open source toolkit for constructing bioinformatics database warehouses using the MySQL and Oracle relational database managers. BioWarehouse integrates its component databases into a common representational framework within a single database management system, thus enabling multi-database queries using the Structured Query Language (SQL) but also facilitating a variety of database integration tasks such as comparative analysis and data mining. BioWarehouse currently supports the integration of a pathway-centric set of databases including ENZYME, KEGG, and BioCyc, and in addition the UniProt, GenBank, NCBI Taxonomy, and CMR databases, and the Gene Ontology. Loader tools, written in the C and JAVA languages, parse and load these databases into a relational database schema. The loaders also apply a degree of semantic normalization to their respective source data, decreasing semantic heterogeneity. The schema supports the following bioinformatics datatypes: chemical compounds, biochemical reactions, metabolic pathways, proteins, genes, nucleic acid sequences, features on protein and nucleic-acid sequences, organisms, organism taxonomies, and controlled vocabularies. As an application example, we applied BioWarehouse to determine the fraction of biochemically characterized enzyme activities for which no sequences exist in the public sequence databases. The answer is that no sequence exists for 36% of enzyme activities for which EC numbers have been assigned. These gaps in sequence data significantly limit the accuracy of genome annotation and metabolic pathway prediction, and are a barrier for metabolic engineering. Complex queries of this type provide examples of the value of the data warehousing approach to bioinformatics research. Conclusion BioWarehouse embodies significant progress on the database integration problem for bioinformatics. PMID:16556315

  3. BioWarehouse: a bioinformatics database warehouse toolkit.

    PubMed

    Lee, Thomas J; Pouliot, Yannick; Wagner, Valerie; Gupta, Priyanka; Stringer-Calvert, David W J; Tenenbaum, Jessica D; Karp, Peter D

    2006-03-23

    This article addresses the problem of interoperation of heterogeneous bioinformatics databases. We introduce BioWarehouse, an open source toolkit for constructing bioinformatics database warehouses using the MySQL and Oracle relational database managers. BioWarehouse integrates its component databases into a common representational framework within a single database management system, thus enabling multi-database queries using the Structured Query Language (SQL) but also facilitating a variety of database integration tasks such as comparative analysis and data mining. BioWarehouse currently supports the integration of a pathway-centric set of databases including ENZYME, KEGG, and BioCyc, and in addition the UniProt, GenBank, NCBI Taxonomy, and CMR databases, and the Gene Ontology. Loader tools, written in the C and JAVA languages, parse and load these databases into a relational database schema. The loaders also apply a degree of semantic normalization to their respective source data, decreasing semantic heterogeneity. The schema supports the following bioinformatics datatypes: chemical compounds, biochemical reactions, metabolic pathways, proteins, genes, nucleic acid sequences, features on protein and nucleic-acid sequences, organisms, organism taxonomies, and controlled vocabularies. As an application example, we applied BioWarehouse to determine the fraction of biochemically characterized enzyme activities for which no sequences exist in the public sequence databases. The answer is that no sequence exists for 36% of enzyme activities for which EC numbers have been assigned. These gaps in sequence data significantly limit the accuracy of genome annotation and metabolic pathway prediction, and are a barrier for metabolic engineering. Complex queries of this type provide examples of the value of the data warehousing approach to bioinformatics research. BioWarehouse embodies significant progress on the database integration problem for bioinformatics.

  4. Rv0004 is a new essential member of the mycobacterial DNA replication machinery.

    PubMed

    Mann, Katherine M; Huang, Deborah L; Hooppaw, Anna J; Logsdon, Michelle M; Richardson, Kirill; Lee, Hark Joon; Kimmey, Jacqueline M; Aldridge, Bree B; Stallings, Christina L

    2017-11-01

    DNA replication is fundamental for life, yet a detailed understanding of bacterial DNA replication is limited outside the organisms Escherichia coli and Bacillus subtilis. Many bacteria, including mycobacteria, encode no identified homologs of helicase loaders or regulators of the initiator protein DnaA, despite these factors being essential for DNA replication in E. coli and B. subtilis. In this study we discover that a previously uncharacterized protein, Rv0004, from the human pathogen Mycobacterium tuberculosis is essential for bacterial viability and that depletion of Rv0004 leads to a block in cell cycle progression. Using a combination of genetic and biochemical approaches, we found that Rv0004 has a role in DNA replication, interacts with DNA and the replicative helicase DnaB, and affects DnaB-DnaA complex formation. We also identify a conserved domain in Rv0004 that is predicted to structurally resemble the N-terminal protein-protein interaction domain of DnaA. Mutation of a single conserved tryptophan within Rv0004's DnaA N-terminal-like domain leads to phenotypes similar to those observed upon Rv0004 depletion and can affect the association of Rv0004 with DnaB. In addition, using live cell imaging during depletion of Rv0004, we have uncovered a previously unappreciated role for DNA replication in coordinating mycobacterial cell division and cell size. Together, our data support that Rv0004 encodes a homolog of the recently identified DciA family of proteins found in most bacteria that lack the DnaC-DnaI helicase loaders in E. coli and B. subtilis. Therefore, the mechanisms of Rv0004 elucidated here likely apply to other DciA homologs and reveal insight into the diversity of bacterial strategies in even the most conserved biological processes.

  5. Cytology of the minor-vein phloem in 320 species from the subclass Asteridae suggests a high diversity of phloem-loading modes†

    PubMed Central

    Batashev, Denis R.; Pakhomova, Marina V.; Razumovskaya, Anna V.; Voitsekhovskaja, Olga V.; Gamalei, Yuri V.

    2013-01-01

    The discovery of abundant plasmodesmata at the bundle sheath/phloem interface in Oleaceae (Gamalei, 1974) and Cucurbitaceae (Turgeon et al., 1975) raised the questions as to whether these plasmodesmata are functional in phloem loading and how widespread symplasmic loading would be. Analysis of over 800 dicot species allowed the definition of “open” and “closed” types of the minor vein phloem depending on the abundance of plasmodesmata between companion cells and bundle sheath (Gamalei, 1989, 1990). These types corresponded to potential symplasmic and apoplasmic phloem loaders, respectively; however, this definition covered a spectrum of diverse structures of phloem endings. Here, a review of detailed cytological analyses of minor veins in 320 species from the subclass Asteridae is presented, including data on companion cell types and their combinations which have not been reported previously. The percentage of Asteridae species with “open” minor vein cytology which also contain sieve-element-companion cell complexes with “closed” cytology, i.e., that show specialization for both symplasmic and apoplasmic phloem loading, was determined. Along with recent data confirming the dissimilar functional specialization of structurally different parts of minor vein phloem in the stachyose-translocating species Alonsoa meridionalis (Voitsekhovskaja et al., 2009), these findings suggest that apoplasmic loading is indispensable in a large group of species previously classified as putative symplasmic loaders. Altogether, this study provides formal classifications of companion cells and of minor veins, respectively, in 24 families of the Asteridae based on their structural features, opening the way to a close investigation of the relationship between structure and function in phloem loading. PMID:23970890

  6. Structural basis for the unfolding of anthrax lethal factor by protective antigen oligomers

    PubMed Central

    Feld, Geoffrey K.; Thoren, Katie L.; Kintzer, Alexander F.; Sterling, Harry J.; Tang, Iok I.; Greenberg, Shoshana G.; Williams, Evan R.; Krantz, Bryan A.

    2011-01-01

    The protein transporter, anthrax lethal toxin, is comprised of protective antigen (PA), a transmembrane translocase, and lethal factor (LF), a cytotoxic enzyme. Following assembly into holotoxin complexes, PA forms an oligomeric channel that unfolds LF and translocates it into the host cell. We report the crystal structure of the core of a lethal toxin complex to 3.1-Å resolution; the structure contains a PA octamer bound to four LF PA-binding domains (LFN). The first α helix and β strand of each LFN unfold and dock into a deep amphipathic cleft on the surface of the PA octamer, which we call the α clamp. The α clamp possesses nonspecific polypeptide binding activity and is functionally relevant to efficient holotoxin assembly, PA octamer formation, and LF unfolding and translocation. This structure provides insight on the mechanism of translocation-coupled protein unfolding. PMID:21037566

  7. [Modulation of Kv4 channels by KChIPs clamping].

    PubMed

    Cui, Yuan-Yuan; Wang, Ke-Wei

    2009-01-01

    The rapidly inactivating (A-type) potassium channels regulate membrane excitability that defines the fundamental mechanism of neuronal functions such as pain signaling. Cytosolic Kv channel-interacting proteins KChIPs co-assemble with Kv4 (Shal) alpha subunits to form a native complex. The specific binding of auxiliary KChIPs to the Kv4 N-terminus results in modulation of gating properties, surface expression and subunit assembly of Kv4 channels. Based on recent structural efforts, here we attempt to emphasize the interaction between KChIPs and Kv4 channel complex in which a single KChIP1 molecule laterally clamps two neighboring Kv4.3 N-termini in a 4:4 manner. Greater insights into molecular mechanism between KChIPs and Kv4 interaction may provide therapeutic potentials by structure-based design of chemical compounds aimed at disrupting the protein-protein interaction for treatment of membrane excitability-related disorders.

  8. 29 CFR 1926.1001 - Minimum performance criteria for rollover protective structures for designated scrapers, loaders...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... of Steel Products (available at each Regional Office of the Occupational Safety and Health... Society of Automotive Engineers Recommended Practices: SAE J320a, Minimum Performance Criteria for Roll...

  9. 29 CFR 1926.1001 - Minimum performance criteria for rollover protective structures for designated scrapers, loaders...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... of Steel Products (available at each Regional Office of the Occupational Safety and Health... Society of Automotive Engineers Recommended Practices: SAE J320a, Minimum Performance Criteria for Roll...

  10. 29 CFR 1926.1001 - Minimum performance criteria for rollover protective structures for designated scrapers, loaders...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... of Steel Products (available at each Regional Office of the Occupational Safety and Health... Society of Automotive Engineers Recommended Practices: SAE J320a, Minimum Performance Criteria for Roll...

  11. 29 CFR 1926.1001 - Minimum performance criteria for rollover protective structures for designated scrapers, loaders...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... of Steel Products (available at each Regional Office of the Occupational Safety and Health... Society of Automotive Engineers Recommended Practices: SAE J320a, Minimum Performance Criteria for Roll...

  12. 29 CFR 1926.1001 - Minimum performance criteria for rollover protective structures for designated scrapers, loaders...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... of Steel Products (available at each Regional Office of the Occupational Safety and Health... Society of Automotive Engineers Recommended Practices: SAE J320a, Minimum Performance Criteria for Roll...

  13. Cognitive simulation of incident risks in the structure of loading and transport enterprise

    NASA Astrophysics Data System (ADS)

    Shishkina, S. V.; Pristupa, Yu D.; Pavlova, L. D.; Fryanov, V. N.

    2017-09-01

    Organizational and technical system of a manufacturing enterprise was identified, which includes three subsystems: main production, industrial and social infrastructure. Based on the results of cognitive modeling, significant system concepts were identified that reduce the risks of incidents. The internal control influences formed in accordance with level of competence of heads of services, departments, sections, dispatchers, acting on the basis of regulations, job profiles. The second concept influencing the enterprise management system is personnel, which is assessed by the compliance of competencies of crane operators, loader operators, slingers, loaders, and acceptance/delivery agents to job responsibilities and labor functions. At a low level of professional competencies, the personnel does not fully comply with job duties and labor functions, the risk of an incident is maximal. The application of cognitive modeling allows us to identify the essential elements that ensure stable functioning of the system as a whole.

  14. In-line drivetrain and four wheel drive work machine using same

    DOEpatents

    Hoff, Brian

    2008-08-05

    A four wheel drive articulated mine loader is powered by a fuel cell and propelled by a single electric motor. The drivetrain has the first axle, second axle, and motor arranged in series on the work machine chassis. Torque is carried from the electric motor to the back differential via a pinion meshed with the ring gear of the back differential. A second pinion oriented in an opposite direction away from the ring gear is coupled to a drive shaft to transfer torque from the ring gear to the differential of the front axle. Thus, the ring gear of the back differential acts both to receive torque from the motor and to transfer torque to the forward axle. The in-line drive configuration includes a single electric motor and a single reduction gear to power the four wheel drive mine loader.

  15. Is early cord clamping, delayed cord clamping or cord milking best?

    PubMed

    Vatansever, Binay; Demirel, Gamze; Ciler Eren, Elif; Erel, Ozcan; Neselioglu, Salim; Karavar, Hande Nur; Gundogdu, Semra; Ulfer, Gozde; Bahadir, Selcen; Tastekin, Ayhan

    2018-04-01

    To compare the antioxidant status of three cord clamping procedures (early clamping, delayed clamping and milking) by analyzing the thiol-disulfide balance. This randomized controlled study enrolled 189 term infants who were divided into three groups according to the cord clamping procedure: early clamping, delayed clamping and milking. Blood samples were collected from the umbilical arteries immediately after clamping, and the thiol/disulfide homeostasis was analyzed. The native and total thiol levels were significantly (p < .05) lower in the early cord clamping group compared with the other two groups. The disulfide/total thiol ratio was significantly (p = .026) lower in the delayed cord clamping and milking groups compared with the early clamping groups. Early cord clamping causes the production of more disulfide bonds and lower thiol levels, indicating that oxidation reactions are increased in the early cord clamping procedure compared with the delayed cord clamping and milking procedures. The oxidant capacity is greater with early cord clamping than with delayed clamping or cord milking. Delayed cord clamping or milking are beneficial in neonatal care, and we suggest that they be performed routinely in all deliveries.

  16. Remote drill bit loader

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

    Dokos, J.A.

    1996-12-31

    A drill bit loader is described for loading a tapered shank of a drill bit into a similarly tapered recess in the end of a drill spindle. The spindle has a transverse slot at the inner end of the recess. The end of the tapered shank of the drill bit has a transverse tang adapted to engage in the slot so that the drill bit will be rotated by the spindle. The loader is in the form of a cylinder adapted to receive the drill bit with the shank projecting out of the outer end of the cylinder. Retainer pinsmore » prevent rotation of the drill bit in the cylinder. The spindle is lowered to extend the shank of the drill bit into the recess in the spindle and the spindle is rotated to align the slot in the spindle with the tang on the shank. A spring unit in the cylinder is compressed by the drill bit during its entry into the recess of the spindle and resiliently drives the tang into the slot in the spindle when the tang and slot are aligned. In typical remote drilling operations, whether in hot cells or water pits, drill bits have been held using a collet or end mill type holder with set screws. In either case, to load or change a drill bit required the use master-slave manipulators to position the bits and tighten the collet or set screws. This requirement eliminated many otherwise useful work areas because they were not equipped with slaves, particularly in water pits.« less

  17. Extension modules for storage, visualization and querying of genomic, genetic and breeding data in Tripal databases

    PubMed Central

    Lee, Taein; Cheng, Chun-Huai; Ficklin, Stephen; Yu, Jing; Humann, Jodi; Main, Dorrie

    2017-01-01

    Abstract Tripal is an open-source database platform primarily used for development of genomic, genetic and breeding databases. We report here on the release of the Chado Loader, Chado Data Display and Chado Search modules to extend the functionality of the core Tripal modules. These new extension modules provide additional tools for (1) data loading, (2) customized visualization and (3) advanced search functions for supported data types such as organism, marker, QTL/Mendelian Trait Loci, germplasm, map, project, phenotype, genotype and their respective metadata. The Chado Loader module provides data collection templates in Excel with defined metadata and data loaders with front end forms. The Chado Data Display module contains tools to visualize each data type and the metadata which can be used as is or customized as desired. The Chado Search module provides search and download functionality for the supported data types. Also included are the tools to visualize map and species summary. The use of materialized views in the Chado Search module enables better performance as well as flexibility of data modeling in Chado, allowing existing Tripal databases with different metadata types to utilize the module. These Tripal Extension modules are implemented in the Genome Database for Rosaceae (rosaceae.org), CottonGen (cottongen.org), Citrus Genome Database (citrusgenomedb.org), Genome Database for Vaccinium (vaccinium.org) and the Cool Season Food Legume Database (coolseasonfoodlegume.org). Database URL: https://www.citrusgenomedb.org/, https://www.coolseasonfoodlegume.org/, https://www.cottongen.org/, https://www.rosaceae.org/, https://www.vaccinium.org/

  18. Assessment of respirable dust and its free silica contents in different Indian coalmines.

    PubMed

    Mukherjee, Ashit K; Bhattacharya, Sanat K; Saiyed, Habibullah N

    2005-04-01

    Assessment of respirable dust, personal exposures of miners and free silica contents in dust were undertaken to find out the associated risk of coal workers' pneumoconiosis in 9 coal mines of Eastern India during 1988-91. Mine Research Establishment (MRE), 113A Gravimetric Dust Sampler (GDS) and personal samplers (AFC 123), Cassella, London, approved by Director General of Mines Safety (DGMS) were used respectively for monitoring of mine air dust and personal exposures of miners. Fourier Transform Infra-red (FTIR) Spectroscopy determined free silica in respirable dusts. Thermal Conditions like Wet Bulb Globe Temperature (WBGT) index, humidity and wind velocity were also recorded during monitoring. The dust levels in the face return air of both, Board & Pillar (B&P) and Long Wall (LW) mining were found above the permissible level recommended by DGMS, Govt. of India. The drilling, blasting and loading are the major dusty operations in B&P method. Exposures of driller and loader were varied between, 0.81-9.48 mg/m3 and 0.05-9.84 mg/m3 respectively in B&P mining, whereas exposures of DOSCO loader, Shearer operator and Power Support Face Worker were varied between 2.65-9.11 mg/m3, 0.22-10.00 mg/m3 and 0.12-9.32 mg/m3 respectively in LW mining. In open cast mining, compressor and driller operators are the major exposed groups. The percentage silica in respirable dusts found below 5% in all most all the workers except among query loaders and drillers of open cast mines.

  19. A Dynamic Clamp on Every Rig

    PubMed Central

    2017-01-01

    Abstract The dynamic clamp should be a standard part of every cellular electrophysiologist’s toolbox. That it is not, even 25 years after its introduction, comes down to three issues: money, the disruption that adding dynamic clamp to an existing electrophysiology rig entails, and the technical prowess required of experimenters. These have been valid and limiting issues in the past, but no longer. Technological advances associated with the so-called maker movement render them moot. We demonstrate this by implementing a fast (∼100 kHz) dynamic clamp system using an inexpensive microcontroller (Teensy 3.6). The overall cost of the system is less than USD$100, and assembling it requires no prior electronics experience. Modifying it—for example, to add Hodgkin–Huxley-style conductances—requires no prior programming experience. The system works together with existing electrophysiology data acquisition systems (for Macintosh, Windows, and Linux); it does not attempt to supplant them. Moreover, the process of assembling, modifying, and using the system constitutes a useful pedagogical exercise for students and researchers with no background but an interest in electronics and programming. We demonstrate the system’s utility by implementing conductances as fast as a transient sodium conductance and as complex as the Ornstein–Uhlenbeck conductances of the “point conductance” model of synaptic background activity. PMID:29085905

  20. A Structural Basis for the Regulatory Inactivation of DnaA

    PubMed Central

    Xu, Qingping; McMullan, Daniel; Abdubek, Polat; Astakhova, Tamara; Carlton, Dennis; Chen, Connie; Chiu, Hsiu-Ju; Clayton, Thomas; Das, Debanu; Deller, Marc C.; Duan, Lian; Elsliger, Marc-Andre; Feuerhelm, Julie; Hale, Joanna; Han, Gye Won; Jaroszewski, Lukasz; Jin, Kevin K.; Johnson, Hope A.; Klock, Heath E.; Knuth, Mark W.; Kozbial, Piotr; Krishna, S. Sri; Kumar, Abhinav; Marciano, David; Miller, Mitchell D.; Morse, Andrew T.; Nigoghossian, Edward; Nopakun, Amanda; Okach, Linda; Oommachen, Silvya; Paulsen, Jessica; Puckett, Christina; Reyes, Ron; Rife, Christopher L.; Sefcovic, Natasha; Trame, Christine; van den Bedem, Henry; Weekes, Dana; Hodgson, Keith O.; Wooley, John; Deacon, Ashley M.; Godzik, Adam; Lesley, Scott A.; Wilson, Ian A.

    2009-01-01

    Summary Regulatory inactivation of DnaA is dependent on Hda, a protein homologous to the AAA+ ATPase region of the replication initiator DnaA. When bound to the sliding clamp loaded onto duplex DNA, Hda can stimulate the transformation of active DnaA-ATP into inactive DnaA-ADP. The crystal structure of Hda from Shewanella amazonensis SB2B at 1.75 Å resolution reveals that Hda resembles typical AAA+ ATPases. The arrangement of the two subdomains in Hda (residues 1-174, 175-241) differs dramatically from that of DnaA. A CDP molecule anchors the Hda domains in a conformation which promotes dimer formation. The Hda dimer adopts a novel oligomeric assembly for AAA+ proteins in which the arginine finger, crucial for ATP hydrolysis, is fully exposed and available to hydrolyze DnaA-ATP through a typical AAA+ type mechanism. The sliding clamp binding motifs at the N-terminus of each Hda monomer are partially buried and combine to form an antiparallel β-sheet at the dimer interface. The inaccessibility of the clamp binding motifs in the CDP bound structure of Hda suggests that conformational changes are required for Hda to form a functional complex with the clamp. Thus, the CDP-bound Hda dimer likely represents an inactive form of Hda. PMID:19000695

  1. --No Title--

    Science.gov Websites

    ;background:url(/technical-assistance/assets/images/ajax-loader.gif) center center no-repeat}.btn-long{font-size :0 0 10px}.btn-long{height:80px}}@media only screen and (min-width :1200px){.btn-long{height:60px

  2. PRN 2009-1: Establishment of Antimicrobial Exposure Assessment Task Force II

    EPA Pesticide Factsheets

    This PR notice announces the establishment of the Antimicrobial Exposure Assessment Task Force II, an industry-wide task force to develop mixer, loader, applicator and post-application exposure data for antimicrobial pesticides used in various settings.

  3. 14 CFR 158.3 - Definitions.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Definitions. 158.3 Section 158.3 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) AIRPORTS... airport to support aeronautical operations and related activities. Baggage tugs, belt loaders, cargo...

  4. 14 CFR 158.3 - Definitions.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Definitions. 158.3 Section 158.3 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) AIRPORTS... airport to support aeronautical operations and related activities. Baggage tugs, belt loaders, cargo...

  5. Influence of clamping plate permeability and metal screen structures on three-dimensional magnetic field and eddy current loss in end region of a turbo-generator by numerical analysis

    NASA Astrophysics Data System (ADS)

    Likun, Wang; Weili, Li; Yi, Xue; Chunwei, Guan

    2013-11-01

    A significant problem of turbogenerators on complex end structures is overheating of local parts caused by end losses in the end region. Therefore, it is important to investigate the 3-D magnetic field and eddy current loss in the end. In end region of operating large turbogenerator at thermal power plants, magnetic leakage field distribution is complex. In this paper, a 3-D mathematical model used for the calculation of the electromagnetic field in the end region of large turbo-generators is given. The influence of spatial locations of end structures, the actual shape and material of end windings, clamping plate, and copper screen are considered. Adopting the time-step finite element (FE) method and taking the nonlinear characteristics of the core into consideration, a 3-D transient magnetic field is calculated. The objective of this paper is to investigate the influence of clamping plate permeability and metal screen structures on 3-D electromagnetic field distribution and eddy current loss in end region of a turbo-generator. To reduce the temperature of copper screen, a hollow metal screen is proposed. The eddy current loss, which is gained from the 3D transient magnetic field, is used as heat source for the thermal field of end region. The calculated temperatures are compared with test data.

  6. Mutant DnaAs of Escherichia coli that are refractory to negative control

    PubMed Central

    Chodavarapu, Sundari; Felczak, Magdalena M.; Simmons, Lyle A.; Murillo, Alec; Kaguni, Jon M.

    2013-01-01

    DnaA is the initiator of DNA replication in bacteria. A mutant DnaA named DnaAcos is unusual because it is refractory to negative regulation. We developed a genetic method to isolate other mutant DnaAs that circumvent regulation to extend our understanding of mechanisms that control replication initiation. Like DnaAcos, one mutant bearing a tyrosine substitution for histidine 202 (H202Y) withstands the regulation exerted by datA, hda and dnaN (β clamp), and both DnaAcos and H202Y resist inhibition by the Hda-β clamp complex in vitro. Other mutant DnaAs carrying G79D, E244K, V303M or E445K substitutions are either only partially sensitive or refractory to inhibition by the Hda-β clamp complex in vitro but are responsive to hda expression in vivo. All mutant DnaAs remain able to interact directly with Hda. Of interest, both DnaAcos and DnaAE244K bind more avidly to Hda. These mutants, by sequestrating Hda, may limit its availability to regulate other DnaA molecules, which remain active to induce extra rounds of DNA replication. Other evidence suggests that a mutant bearing a V292M substitution hyperinitiates by escaping the effect of an unknown regulatory factor. Together, our results provide new insight into the mechanisms that regulate replication initiation in Escherichia coli. PMID:23990329

  7. Mutant DnaAs of Escherichia coli that are refractory to negative control.

    PubMed

    Chodavarapu, Sundari; Felczak, Magdalena M; Simmons, Lyle A; Murillo, Alec; Kaguni, Jon M

    2013-12-01

    DnaA is the initiator of DNA replication in bacteria. A mutant DnaA named DnaAcos is unusual because it is refractory to negative regulation. We developed a genetic method to isolate other mutant DnaAs that circumvent regulation to extend our understanding of mechanisms that control replication initiation. Like DnaAcos, one mutant bearing a tyrosine substitution for histidine 202 (H202Y) withstands the regulation exerted by datA, hda and dnaN (β clamp), and both DnaAcos and H202Y resist inhibition by the Hda-β clamp complex in vitro. Other mutant DnaAs carrying G79D, E244K, V303M or E445K substitutions are either only partially sensitive or refractory to inhibition by the Hda-β clamp complex in vitro but are responsive to hda expression in vivo. All mutant DnaAs remain able to interact directly with Hda. Of interest, both DnaAcos and DnaAE244K bind more avidly to Hda. These mutants, by sequestrating Hda, may limit its availability to regulate other DnaA molecules, which remain active to induce extra rounds of DNA replication. Other evidence suggests that a mutant bearing a V292M substitution hyperinitiates by escaping the effect of an unknown regulatory factor. Together, our results provide new insight into the mechanisms that regulate replication initiation in Escherichia coli.

  8. PRN 2007-3: The Agricultural Handlers Exposure Task Force, L.L.C

    EPA Pesticide Factsheets

    This PR Notice discusses the Agricultural Handlers Exposure Task Force, an industry-wide task force formed to develop mixer, loader, and applicator exposure data for pesticides used in agricultural settings. It includes contacts for more information.

  9. Triple bar, high efficiency mechanical sealer

    DOEpatents

    Pak, Donald J.; Hawkins, Samantha A.; Young, John E.

    2013-03-19

    A clamp with a bottom clamp bar that has a planar upper surface is provided. The clamp may also include a top clamp bar connected to the bottom clamp bar, and a pressure distribution bar between the top clamp bar and the bottom clamp bar. The pressure distribution bar may have a planar lower surface in facing relation to the upper surface of the bottom clamp bar. An object is capable of being disposed in a clamping region between the upper surface and the lower surface. The width of the planar lower surface may be less than the width of the upper surface within the clamping region. Also, the pressure distribution bar may be capable of being urged away from the top clamp bar and towards the bottom clamp bar.

  10. Drosophila model of Meier-Gorlin syndrome based on the mutation in a conserved C-Terminal domain of Orc6.

    PubMed

    Balasov, Maxim; Akhmetova, Katarina; Chesnokov, Igor

    2015-11-01

    Meier-Gorlin syndrome (MGS) is an autosomal recessive disorder characterized by microtia, primordial dwarfism, small ears, and skeletal abnormalities. Patients with MGS often carry mutations in the genes encoding the components of the pre-replicative complex such as Origin Recognition Complex (ORC) subunits Orc1, Orc4, Orc6, and helicase loaders Cdt1 and Cdc6. Orc6 is an important component of ORC and has functions in both DNA replication and cytokinesis. Mutation in conserved C-terminal motif of Orc6 associated with MGS impedes the interaction of Orc6 with core ORC. In order to study the effects of MGS mutation in an animal model system we introduced MGS mutation in Orc6 and established Drosophila model of MGS. Mutant flies die at third instar larval stage with abnormal chromosomes and DNA replication defects. The lethality can be rescued by elevated expression of mutant Orc6 protein. Rescued MGS flies are unable to fly and display multiple planar cell polarity defects. © 2015 Wiley Periodicals, Inc. © 2015 Wiley Periodicals, Inc.

  11. Post clamp

    NASA Technical Reports Server (NTRS)

    Ramsey, John K. (Inventor); Meyn, Erwin H. (Inventor)

    1990-01-01

    A pair of spaced collars are mounted at right angles on a clamp body by retaining rings which enable the collars to rotate with respect to the clamp body. Mounting posts extend through aligned holes in the collars and clamp body. Each collar can be clamped onto the inserted post while the clamp body remains free to rotate about the post and collar. The clamp body is selectively clamped onto each post.

  12. Complex Regional Pain Syndrome

    MedlinePlus

    ... an anesthetic (pain reliever) into certain nerves. This blocks the pain signals. If the injection relieves the pain, it may be repeated. It is not a cure. Sympathectomy of the injured nerve. A surgeon will cut or clamp the nerve chain. This has been reported to improve pain caused ...

  13. Modulation of the Pyrococcus abyssi NucS Endonuclease Activity by Replication Clamp at Functional and Structural Levels*

    PubMed Central

    Creze, Christophe; Ligabue, Alessio; Laurent, Sébastien; Lestini, Roxane; Laptenok, Sergey P.; Khun, Joelle; Vos, Marten H.; Czjzek, Mirjam; Myllykallio, Hannu; Flament, Didier

    2012-01-01

    Pyrococcus abyssi NucS is the founding member of a new family of structure-specific DNA endonucleases that interact with the replication clamp proliferating cell nuclear antigen (PCNA). Using a combination of small angle x-ray scattering and surface plasmon resonance analyses, we demonstrate the formation of a stable complex in solution, in which one molecule of the PabNucS homodimer binds to the outside surface of the PabPCNA homotrimer. Using fluorescent labels, PCNA is shown to increase the binding affinity of NucS toward single-strand/double-strand junctions on 5′ and 3′ flaps, as well as to modulate the cleavage specificity on the branched DNA structures. Our results indicate that the presence of a single major contact between the PabNucS and PabPCNA proteins, together with the complex-induced DNA bending, facilitate conformational flexibility required for specific cleavage at the single-strand/double-strand DNA junction. PMID:22431731

  14. Modulation of the Pyrococcus abyssi NucS endonuclease activity by replication clamp at functional and structural levels.

    PubMed

    Creze, Christophe; Ligabue, Alessio; Laurent, Sébastien; Lestini, Roxane; Laptenok, Sergey P; Khun, Joelle; Vos, Marten H; Czjzek, Mirjam; Myllykallio, Hannu; Flament, Didier

    2012-05-04

    Pyrococcus abyssi NucS is the founding member of a new family of structure-specific DNA endonucleases that interact with the replication clamp proliferating cell nuclear antigen (PCNA). Using a combination of small angle x-ray scattering and surface plasmon resonance analyses, we demonstrate the formation of a stable complex in solution, in which one molecule of the PabNucS homodimer binds to the outside surface of the PabPCNA homotrimer. Using fluorescent labels, PCNA is shown to increase the binding affinity of NucS toward single-strand/double-strand junctions on 5' and 3' flaps, as well as to modulate the cleavage specificity on the branched DNA structures. Our results indicate that the presence of a single major contact between the PabNucS and PabPCNA proteins, together with the complex-induced DNA bending, facilitate conformational flexibility required for specific cleavage at the single-strand/double-strand DNA junction.

  15. Numerical assessment of fore-and-aft suspension performance to reduce whole-body vibration of wheel loader drivers

    NASA Astrophysics Data System (ADS)

    Fleury, Gérard; Mistrot, Pierre

    2006-12-01

    While driving off-road vehicles, operators are exposed to whole-body vibration acting in the fore-and-aft direction. Seat manufacturers supply products equipped with fore-and-aft suspension but only a few studies report on their performance. This work proposes a computational approach to design fore-and-aft suspensions for wheel loader seats. Field tests were conducted in a quarry to analyse the nature of vibration to which the driver was exposed. Typical input signals were recorded to be reproduced in the laboratory. Technical specifications are defined for the suspension. In order to evaluate the suspension vibration attenuation performance, a model of a sitting human body was developed and coupled to a seat model. The seat model combines the models of each suspension component. A linear two-degree-of-freedom model is used to describe the dynamic behaviour of the sitting driver. Model parameters are identified by fitting the computed apparent mass frequency response functions to the measured values. Model extensions are proposed to investigate postural effects involving variations in hands and feet positions and interaction of the driver's back with the backrest. Suspension design parameters are firstly optimized by computing the seat/man model response to sinusoidal acceleration. Four criteria including transmissibility, interaction force between the driver's back and the backrest and relative maximal displacement of the suspension are computed. A new suspension design with optimized features is proposed. Its performance is checked from calculations of the response of the seat/man model subjected to acceleration measured on the wheel loader during real work conditions. On the basis of the computed values of the SEAT factors, it is found possible to design a suspension that would increase the attenuation provided by the seat by a factor of two.

  16. Is increased joint loading detrimental to obese patients with knee osteoarthritis? A secondary data analysis from a randomized trial.

    PubMed

    Henriksen, M; Hunter, D J; Dam, E B; Messier, S P; Andriacchi, T P; Lohmander, L S; Aaboe, J; Boesen, M; Gudbergsen, H; Bliddal, H; Christensen, R

    2013-12-01

    To investigate whether increased knee joint loading due to improved ambulatory function and walking speed following weight loss achieved over 16 weeks accelerates symptomatic and structural disease progression over a subsequent 1 year weight maintenance period in an obese population with knee osteoarthritis (OA). Data from a prospective study of weight loss in obese patients with knee OA (the CARtilage in obese knee OsteoarThritis (CAROT) study) were used to determine changes in knee joint compressive loadings (model estimated) during walking after a successful 16 week weight loss intervention. The participants were divided into 'Unloaders' (participants that reduced joint loads) and 'Loaders' (participants that increased joint loads). The primary symptomatic outcome was changes in knee symptoms, measured with the Knee injury and Osteoarthritis Outcome Score (KOOS) questionnaire, during a subsequent 52 weeks weight maintenance period. The primary structural outcome was changes in tibiofemoral cartilage loss assessed semi-quantitatively (Boston Leeds Knee Osteoarthritis Score (BLOKS) from MRI after the 52 weight maintenance period. 157 participants (82% of the CAROT cohort) with medial and/or lateral knee OA were classified as Unloaders (n = 100) or Loaders (n = 57). The groups showed similar significant changes in symptoms (group difference: -2.4 KOOS points [95% CI -6.8:1.9]) and cartilage loss (group difference: -0.06 BLOKS points [95% CI -0.22:0.11) after 1 year, with no statistically significant differences between Loaders and Unloaders. For obese patients undergoing a significant weight loss, increased knee joint loading for 1 year was not associated with accelerated symptomatic and structural disease progression compared to a similar weight loss group that had reduced ambulatory compressive knee joint loads. NCT00655941. Copyright © 2013 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

  17. Use of the Satinsky clamp for hilar clamping during robotic partial nephrectomy: indications, technique, and multi-center outcomes.

    PubMed

    Abdullah, Newaj; Rahbar, Haider; Barod, Ravi; Dalela, Deepansh; Larson, Jeff; Johnson, Michael; Mass, Alon; Zargar, Homayoun; Kaouk, Jihad; Allaf, Mohamad; Bhayani, Sam; Stifelman, Michael; Rogers, Craig

    2017-03-01

    A Satinsky clamp may be a backup option for hilar clamping during robotic partial nephrectomy (RPN) if there are challenges with application of bulldog clamps, but there are potential safety concerns. We evaluate outcomes of RPN using Satinsky vs. bulldog clamps, and provide tips for safe use of the Satinsky as a backup option. Using a multi-center database, we identified 1073 patients who underwent RPN between 2006 and 2013, and had information available about method of hilar clamping (bulldog clamp vs. Satinsky clamp). Patient baseline characteristics, tumor features, and perioperative outcomes were compared between the Satinsky and bulldog clamp groups. A Satinsky clamp was used for hilar clamping in 94 (8.8 %) RPN cases, and bulldog clamps were used in 979 (91.2 %) cases. The use of a Satinsky clamp was associated with greater operative time (198 vs. 175 min, p < 0.001), estimated blood loss (EBL, 200 vs. 100 ml, p < 0.001), warm ischemia time (WIT, 20 vs. 19 min, p = 0.036), transfusion rate (12.8 vs. 4.8 %, p = 0.001), and hospital stay (3 vs. 2 days, p < 0.001). Tumor characteristics and number of renal vessels were similar between groups. There were six intraoperative complications in the Satinsky clamp group, but none were directly related to the Satinsky clamp. On multivariable analysis, the use of the Satinsky clamp was not associated with increase in intraoperative or Clavien ≥3 postoperative complications, positive surgical margin rate or percentage change in estimated glomerular filtration rate. A Satinsky clamp can be a backup option for hilar clamping during challenging RPN cases, but requires careful technique, and was rarely necessary.

  18. Effect of sterilization on stiffness and dimensional stability of rubber-dam clamps.

    PubMed

    Giebink, D L; Mathieu, G P; Hondrum, S O

    1996-01-01

    Simulated clinical conditions were used to test the effect of sterilization on rubber-dam clamp stiffness and dimension. Sixty Hygienic and Ivory W7 clamps were either steam or dry heat sterilized and compared to controls. Stiffness and dimensional change between Ivory clamp groups was significant (p<.0001); the sterilized clamps showed less change than the controls. Hygienic groups showed a significant different between the control and dry heat groups (p<.05); the sterilized clamps showed less change than the controls. The change in stiffness and interjaw width for all Ivory clamps compared to all Hygienic clamps was significant (p<.0001). The Hygienic clamps changes less than the Ivory clamps. The results indicate that steam and dry heat sterilization do not affect retention of rubber-dam clamps.

  19. A structural basis for the regulatory inactivation of DnaA.

    PubMed

    Xu, Qingping; McMullan, Daniel; Abdubek, Polat; Astakhova, Tamara; Carlton, Dennis; Chen, Connie; Chiu, Hsiu-Ju; Clayton, Thomas; Das, Debanu; Deller, Marc C; Duan, Lian; Elsliger, Marc-Andre; Feuerhelm, Julie; Hale, Joanna; Han, Gye Won; Jaroszewski, Lukasz; Jin, Kevin K; Johnson, Hope A; Klock, Heath E; Knuth, Mark W; Kozbial, Piotr; Sri Krishna, S; Kumar, Abhinav; Marciano, David; Miller, Mitchell D; Morse, Andrew T; Nigoghossian, Edward; Nopakun, Amanda; Okach, Linda; Oommachen, Silvya; Paulsen, Jessica; Puckett, Christina; Reyes, Ron; Rife, Christopher L; Sefcovic, Natasha; Trame, Christine; van den Bedem, Henry; Weekes, Dana; Hodgson, Keith O; Wooley, John; Deacon, Ashley M; Godzik, Adam; Lesley, Scott A; Wilson, Ian A

    2009-01-16

    Regulatory inactivation of DnaA is dependent on Hda (homologous to DnaA), a protein homologous to the AAA+ (ATPases associated with diverse cellular activities) ATPase region of the replication initiator DnaA. When bound to the sliding clamp loaded onto duplex DNA, Hda can stimulate the transformation of active DnaA-ATP into inactive DnaA-ADP. The crystal structure of Hda from Shewanella amazonensis SB2B at 1.75 A resolution reveals that Hda resembles typical AAA+ ATPases. The arrangement of the two subdomains in Hda (residues 1-174 and 175-241) differs dramatically from that of DnaA. A CDP molecule anchors the Hda domains in a conformation that promotes dimer formation. The Hda dimer adopts a novel oligomeric assembly for AAA+ proteins in which the arginine finger, crucial for ATP hydrolysis, is fully exposed and available to hydrolyze DnaA-ATP through a typical AAA+ type of mechanism. The sliding clamp binding motifs at the N-terminus of each Hda monomer are partially buried and combine to form an antiparallel beta-sheet at the dimer interface. The inaccessibility of the clamp binding motifs in the CDP-bound structure of Hda suggests that conformational changes are required for Hda to form a functional complex with the clamp. Thus, the CDP-bound Hda dimer likely represents an inactive form of Hda.

  20. FPGA in-the-loop simulations of cardiac excitation model under voltage clamp conditions

    NASA Astrophysics Data System (ADS)

    Othman, Norliza; Adon, Nur Atiqah; Mahmud, Farhanahani

    2017-01-01

    Voltage clamp technique allows the detection of single channel currents in biological membranes in identifying variety of electrophysiological problems in the cellular level. In this paper, a simulation study of the voltage clamp technique has been presented to analyse current-voltage (I-V) characteristics of ion currents based on Luo-Rudy Phase-I (LR-I) cardiac model by using a Field Programmable Gate Array (FPGA). Nowadays, cardiac models are becoming increasingly complex which can cause a vast amount of time to run the simulation. Thus, a real-time hardware implementation using FPGA could be one of the best solutions for high-performance real-time systems as it provides high configurability and performance, and able to executes in parallel mode operation. For shorter time development while retaining high confidence results, FPGA-based rapid prototyping through HDL Coder from MATLAB software has been used to construct the algorithm for the simulation system. Basically, the HDL Coder is capable to convert the designed MATLAB Simulink blocks into hardware description language (HDL) for the FPGA implementation. As a result, the voltage-clamp fixed-point design of LR-I model has been successfully conducted in MATLAB Simulink and the simulation of the I-V characteristics of the ionic currents has been verified on Xilinx FPGA Virtex-6 XC6VLX240T development board through an FPGA-in-the-loop (FIL) simulation.

  1. Real world duty cycles and utilization for construction equipment in North Carolina.

    DOT National Transportation Integrated Search

    2008-01-04

    Field data for in-use fuel consumption and emission rates were collected for 15 nonroad vehicles using a portable emission : measurement system (PEMS). Each vehicle, including 5 backhoes, 4 front end loaders, and 6 motor graders, were tested : once o...

  2. Comparison of simulator fidelity model predictions with in-simulator evaluation data

    NASA Technical Reports Server (NTRS)

    Parrish, R. V.; Mckissick, B. T.; Ashworth, B. R.

    1983-01-01

    A full factorial in simulator experiment of a single axis, multiloop, compensatory pitch tracking task is described. The experiment was conducted to provide data to validate extensions to an analytic, closed loop model of a real time digital simulation facility. The results of the experiment encompassing various simulation fidelity factors, such as visual delay, digital integration algorithms, computer iteration rates, control loading bandwidths and proprioceptive cues, and g-seat kinesthetic cues, are compared with predictions obtained from the analytic model incorporating an optimal control model of the human pilot. The in-simulator results demonstrate more sensitivity to the g-seat and to the control loader conditions than were predicted by the model. However, the model predictions are generally upheld, although the predicted magnitudes of the states and of the error terms are sometimes off considerably. Of particular concern is the large sensitivity difference for one control loader condition, as well as the model/in-simulator mismatch in the magnitude of the plant states when the other states match.

  3. Volvo CE to expand driveline components and introduce new loader concept at bauma

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

    NONE

    2007-04-15

    This suppliers news report includes a description of Volvo CE's Graphic wheel loader which uses an electric hybrid power sources and is almost emissions free. Simulator training technology is proving its usefulness in establishing higher safety levels in the mining industry. Immersive Technologies Advanced Equipment training simulators are being used to train students at the Academy for Mine Training and Energy Technology in West Virginia. Geo-Explorer core drills manufactured by Multi-Power Products Ltd. of Canada have been purchased by the Pan African Mining Corp. One is to be assigned to the company's coal program where an extensive exploration program ismore » underway in the Sakoa area in Madagascar. A joint venture agreement has been announced between iPackets International and China's Henan YongAn Investment Guarantee Co. and China Coal Information Institute. Under the name GuoAn Mine Safety Communication System, iPacket's wireless communications software and equipment will be deployed in China's mining industry. 3 photos.« less

  4. Assessment of whole-body vibration exposures and influencing factors for quarry haul truck drivers and loader operators

    PubMed Central

    Mayton, Alan G.; Jobes, Christopher C.; Gallagher, Sean

    2015-01-01

    To further assess vibration exposure on haul trucks (HTs) and front-end wheel loaders (FELs), follow-up investigations were conducted at two US crushed stone operations. The purpose was to: 1) evaluate factors such as load/no-load conditions, speed, load capacity, vehicle age, and seat transmissibility relative to vibration exposure; 2) compare exposure levels with existing ISO/ANSI and EUGPG guidelines. Increasing HT speed increased recorded vibration at the chassis and seat as expected. Neither vehicle load nor vehicle speed increased transmissibility. Increasing HT size and age did show transmissibility decreasing. HT dominant-axis wRMS levels (most often the y-axis, lateral or side-to-side direction) were predominantly within the health guidance caution zone (HGCZ). However, several instances showed vibration dose value (VDV) above the exposure limit value (ELV) for the ISO/ANSI guidelines. VDV levels (all dominant x-axis or fore-aft) were within and above the HGCZ for the EUGPG and above the HGCZ for ISO/ANSI guidelines. PMID:26361493

  5. Design and performance tests of a distributed power-driven wheel loader

    NASA Astrophysics Data System (ADS)

    Jin, Xiaolin; Shi, Laide; Bian, Yongming

    2010-03-01

    An improved ZLM15B distributed power-driven wheel loader was designed, whose travel and brake system was accomplished by two permanent magnet synchronous motorized-wheels instead of traditional mechanical components, and whose hydraulic systems such as the working device system and steering system were both actuated by an induction motor. All above systems were flexibly coupled with 3-phase 380VAC electric power with which the diesel engine power is replaced. On the level cement road, traveling, braking, traction and steering tests were carried out separately under non-load and heavy-load conditions. Data show that machine speed is 5 km/h around and travel efficiency of motorized-wheels is above 95%; that machine braking deceleration is between 0.5 and 0.64 m/s2 but efficiency of motorized-wheels is less than 10%; that maximum machine traction is above 2t while efficiency of motorized-wheels is more than 90% and that adaptive differential steering can be smoothly achieved by motorized-wheels.

  6. Design and performance tests of a distributed power-driven wheel loader

    NASA Astrophysics Data System (ADS)

    Jin, Xiaolin; Shi, Laide; Bian, Yongming

    2009-12-01

    An improved ZLM15B distributed power-driven wheel loader was designed, whose travel and brake system was accomplished by two permanent magnet synchronous motorized-wheels instead of traditional mechanical components, and whose hydraulic systems such as the working device system and steering system were both actuated by an induction motor. All above systems were flexibly coupled with 3-phase 380VAC electric power with which the diesel engine power is replaced. On the level cement road, traveling, braking, traction and steering tests were carried out separately under non-load and heavy-load conditions. Data show that machine speed is 5 km/h around and travel efficiency of motorized-wheels is above 95%; that machine braking deceleration is between 0.5 and 0.64 m/s2 but efficiency of motorized-wheels is less than 10%; that maximum machine traction is above 2t while efficiency of motorized-wheels is more than 90% and that adaptive differential steering can be smoothly achieved by motorized-wheels.

  7. Clamping characteristics study on different types of clamping unit

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

    Jiao, Zhiwei; Liu, Haichao; Xie, Pengcheng

    2015-05-22

    Plastic products are becoming more and more widely used in aerospace, IT, digital electronics and many other fields. With the development of technology, the requirement of product precision is getting higher and higher. However, type and working performance of clamping unit play a decisive role in product precision. Clamping characteristics of different types of clamping unit are discussed in this article, which use finite element numerical analysis method through the software ABAQUS to study the clamping uniformity, and detect the clamping force repeatability precision. The result shows that compared with toggled three-platen clamping unit, clamping characteristics of internal circulation two-platenmore » clamping unit are better, for instance, its mold cavity deformation and force that bars and mold parting surface suffered are more uniform, and its clamping uniformity and repeatability precision is also better.« less

  8. An optogenetics- and imaging-assisted simultaneous multiple patch-clamp recording system for decoding complex neural circuits

    PubMed Central

    Wang, Guangfu; Wyskiel, Daniel R; Yang, Weiguo; Wang, Yiqing; Milbern, Lana C; Lalanne, Txomin; Jiang, Xiaolong; Shen, Ying; Sun, Qian-Quan; Zhu, J Julius

    2015-01-01

    Deciphering neuronal circuitry is central to understanding brain function and dysfunction, yet it remains a daunting task. To facilitate the dissection of neuronal circuits, a process requiring functional analysis of synaptic connections and morphological identification of interconnected neurons, we present here a method for stable simultaneous octuple patch-clamp recordings. This method allows physiological analysis of synaptic interconnections among 4–8 simultaneously recorded neurons and/or 10–30 sequentially recorded neurons, and it allows anatomical identification of >85% of recorded interneurons and >99% of recorded principal neurons. We describe how to apply the method to rodent tissue slices; however, it can be used on other model organisms. We also describe the latest refinements and optimizations of mechanics, electronics, optics and software programs that are central to the realization of a combined single- and two-photon microscopy–based, optogenetics- and imaging-assisted, stable, simultaneous quadruple–viguple patch-clamp recording system. Setting up the system, from the beginning of instrument assembly and software installation to full operation, can be completed in 3–4 d. PMID:25654757

  9. Vps39 Interacts with Tom40 to Establish One of Two Functionally Distinct Vacuole-Mitochondria Contact Sites.

    PubMed

    González Montoro, Ayelén; Auffarth, Kathrin; Hönscher, Carina; Bohnert, Maria; Becker, Thomas; Warscheid, Bettina; Reggiori, Fulvio; van der Laan, Martin; Fröhlich, Florian; Ungermann, Christian

    2018-06-04

    The extensive subcellular network of membrane contact sites plays central roles in organelle biogenesis and communication, yet the precise contributions of the involved machineries remain largely enigmatic. The yeast vacuole forms a membrane contact site with mitochondria, called vacuolar and mitochondrial patch (vCLAMP). Formation of vCLAMPs involves the vacuolar Rab GTPase Ypt7 and the Ypt7-interacting Vps39 subunit of the HOPS tethering complex. Here, we uncover the general preprotein translocase of the outer membrane (TOM) subunit Tom40 as the direct binding partner of Vps39 on mitochondria. We identify Vps39 mutants defective in TOM binding, but functional for HOPS. Cells that cannot form vCLAMPs show reduced growth under stress conditions and impaired survival upon starvation. Unexpectedly, our mutant analysis revealed the existence of two functionally independent vacuole-mitochondria MCSs: one formed by the Ypt7-Vps39-Tom40 tether and a second one by Vps13-Mcp1, which is redundant with ER-mitochondrial contacts formed by ERMES. Copyright © 2018 Elsevier Inc. All rights reserved.

  10. Excavating and loading equipment for peat mining

    NASA Astrophysics Data System (ADS)

    Mikhailov, A. V.; Zhigulskaya, A. I.; Yakonovskaya, T. B.

    2017-10-01

    Recently, the issues of sustainable development of Russian regions, related to ensuring energy security, are more urgent than ever. To achieve sustainable development, an integrated approach to the use of local natural resources is needed. Practically in all north regions of the Russian Federation, peat as a local natural resource is widespread, which has a practical application in the area of housing services. The paper presents the evaluation of technologies for open-pit peat mining, as well as analysis of technological equipment for peat production. Special attention is paid to a question of peat materials excavating and loading. The problem of equipment selection in a peat surface mine is complex. Many features, restrictions and criteria need to be considered. Use of low and ultra-low ground pressure excavators and low ground pressure front-end loaders with full-range tires to provide the necessary floatation in the peat bog environment is offered.

  11. Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system

    PubMed Central

    Fujiwara, Kei; Katayama, Tsutomu; Nomura, Shin-ichiro M.

    2013-01-01

    Replication of all living cells relies on the multirounds flow of the central dogma. Especially, expression of DNA replication proteins is a key step to circulate the processes of the central dogma. Here we achieved the entire sequential transcription–translation–replication process by autonomous expression of chromosomal DNA replication machineries from a reconstituted transcription–translation system (PURE system). We found that low temperature is essential to express a complex protein, DNA polymerase III, in a single tube using the PURE system. Addition of the 13 genes, encoding initiator, DNA helicase, helicase loader, RNA primase and DNA polymerase III to the PURE system gave rise to a DNA replication system by a coupling manner. An artificial genetic circuit demonstrated that the DNA produced as a result of the replication is able to provide genetic information for proteins, indicating the in vitro central dogma can sequentially undergo two rounds. PMID:23737447

  12. Atomistic Simulations of Complex DNA DSBs and the Interactions with Ku70/80 Heterodimer

    NASA Technical Reports Server (NTRS)

    Hu, Shaowen; Cucinotta, Francis A.

    2011-01-01

    Compared to DNA with simple DSBs, the complex lesions can enhance the hydrogen bonds opening rate at the DNA terminus, and increase the mobility of the whole duplex. Binding of Ku drastically reduces the structural disruption and flexibility caused by the complex lesions. In all complex DSBs systems, the binding of DSB terminus with Ku70 is softened while the binding of the middle duplex with Ku80 is tightened. Binding of Ku promotes the rigidity of DNA duplexes, due to the clamp structure of the inner surface of the rings of Ku70/80.

  13. The 1990-1991 project summaries

    NASA Technical Reports Server (NTRS)

    1991-01-01

    Project summaries for 1990-91 at the Georgia Institute of Technology are presented. The following research projects were studied: a lunar surface vehicle model; lunar loader/transporter; trenching and cable-laying device for the lunar surface; a lunar vehicle system for habitat transport and placement; and lunar storage facility.

  14. Non-accident release of hazmat from railroad tank cars : training issues

    DOT National Transportation Integrated Search

    1999-07-01

    This report presents the results of a study to determine the extent to which written materials used in training railroad tank car HAZMAT loaders are consistent with the reading skills of the trainees. Data from four case study sites was used to condu...

  15. 30 CFR 56.14106 - Falling object protection.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 30 Mineral Resources 1 2014-07-01 2014-07-01 false Falling object protection. 56.14106 Section 56... Equipment Safety Devices and Maintenance Requirements § 56.14106 Falling object protection. (a) Fork-lift trucks, front-end loaders, and bulldozers shall be provided with falling object protective structures if...

  16. 30 CFR 57.14106 - Falling object protection.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 30 Mineral Resources 1 2013-07-01 2013-07-01 false Falling object protection. 57.14106 Section 57... Equipment Safety Devices and Maintenance Requirements § 57.14106 Falling object protection. (a) Fork-lift trucks, front-end loaders, and bulldozers shall be provided with falling object protective structures if...

  17. 30 CFR 56.14106 - Falling object protection.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 30 Mineral Resources 1 2013-07-01 2013-07-01 false Falling object protection. 56.14106 Section 56... Equipment Safety Devices and Maintenance Requirements § 56.14106 Falling object protection. (a) Fork-lift trucks, front-end loaders, and bulldozers shall be provided with falling object protective structures if...

  18. 30 CFR 57.14106 - Falling object protection.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 30 Mineral Resources 1 2011-07-01 2011-07-01 false Falling object protection. 57.14106 Section 57... Equipment Safety Devices and Maintenance Requirements § 57.14106 Falling object protection. (a) Fork-lift trucks, front-end loaders, and bulldozers shall be provided with falling object protective structures if...

  19. 30 CFR 56.14106 - Falling object protection.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 30 Mineral Resources 1 2012-07-01 2012-07-01 false Falling object protection. 56.14106 Section 56... Equipment Safety Devices and Maintenance Requirements § 56.14106 Falling object protection. (a) Fork-lift trucks, front-end loaders, and bulldozers shall be provided with falling object protective structures if...

  20. 30 CFR 57.14106 - Falling object protection.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Falling object protection. 57.14106 Section 57... Equipment Safety Devices and Maintenance Requirements § 57.14106 Falling object protection. (a) Fork-lift trucks, front-end loaders, and bulldozers shall be provided with falling object protective structures if...

  1. 30 CFR 57.14106 - Falling object protection.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 30 Mineral Resources 1 2012-07-01 2012-07-01 false Falling object protection. 57.14106 Section 57... Equipment Safety Devices and Maintenance Requirements § 57.14106 Falling object protection. (a) Fork-lift trucks, front-end loaders, and bulldozers shall be provided with falling object protective structures if...

  2. 30 CFR 57.14106 - Falling object protection.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 30 Mineral Resources 1 2014-07-01 2014-07-01 false Falling object protection. 57.14106 Section 57... Equipment Safety Devices and Maintenance Requirements § 57.14106 Falling object protection. (a) Fork-lift trucks, front-end loaders, and bulldozers shall be provided with falling object protective structures if...

  3. 30 CFR 56.14106 - Falling object protection.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 30 Mineral Resources 1 2011-07-01 2011-07-01 false Falling object protection. 56.14106 Section 56... Equipment Safety Devices and Maintenance Requirements § 56.14106 Falling object protection. (a) Fork-lift trucks, front-end loaders, and bulldozers shall be provided with falling object protective structures if...

  4. 30 CFR 56.14106 - Falling object protection.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Falling object protection. 56.14106 Section 56... Equipment Safety Devices and Maintenance Requirements § 56.14106 Falling object protection. (a) Fork-lift trucks, front-end loaders, and bulldozers shall be provided with falling object protective structures if...

  5. Radiological tele-immersion for next generation networks.

    PubMed

    Ai, Z; Dech, F; Rasmussen, M; Silverstein, J C

    2000-01-01

    Since the acquisition of high-resolution three-dimensional patient images has become widespread, medical volumetric datasets (CT or MR) larger than 100 MB and encompassing more than 250 slices are common. It is important to make this patient-specific data quickly available and usable to many specialists at different geographical sites. Web-based systems have been developed to provide volume or surface rendering of medical data over networks with low fidelity, but these cannot adequately handle stereoscopic visualization or huge datasets. State-of-the-art virtual reality techniques and high speed networks have made it possible to create an environment for clinicians geographically distributed to immersively share these massive datasets in real-time. An object-oriented method for instantaneously importing medical volumetric data into Tele-Immersive environments has been developed at the Virtual Reality in Medicine Laboratory (VRMedLab) at the University of Illinois at Chicago (UIC). This networked-VR setup is based on LIMBO, an application framework or template that provides the basic capabilities of Tele-Immersion. We have developed a modular general purpose Tele-Immersion program that automatically combines 3D medical data with the methods for handling the data. For this purpose a DICOM loader for IRIS Performer has been developed. The loader was designed for SGI machines as a shared object, which is executed at LIMBO's runtime. The loader loads not only the selected DICOM dataset, but also methods for rendering, handling, and interacting with the data, bringing networked, real-time, stereoscopic interaction with radiological data to reality. Collaborative, interactive methods currently implemented in the loader include cutting planes and windowing. The Tele-Immersive environment has been tested on the UIC campus over an ATM network. We tested the environment with 3 nodes; one ImmersaDesk at the VRMedLab, one CAVE at the Electronic Visualization Laboratory (EVL) on east campus, and a CT scan machine in UIC Hospital. CT data was pulled directly from the scan machine to the Tele-Immersion server in our Laboratory, and then the data was synchronously distributed by our Onyx2 Rack server to all the VR setups. Instead of permitting medical volume visualization at one VR device, by combining teleconferencing, tele-presence, and virtual reality, the Tele-Immersive environment will enable geographically distributed clinicians to intuitively interact with the same medical volumetric models, point, gesture, converse, and see each other. This environment will bring together clinicians at different geographic locations to participate in Tele-Immersive consultation and collaboration.

  6. 30 CFR 46.5 - New miner training.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... the recognition and avoidance of electrical hazards and other hazards present at the mine, such as traffic patterns and control, mobile equipment (e.g., haul trucks and front-end loaders), and loose or... aspects of an assigned task in paragraph (b)(4) of this section, if hazard recognition training specific...

  7. 30 CFR 46.5 - New miner training.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... the recognition and avoidance of electrical hazards and other hazards present at the mine, such as traffic patterns and control, mobile equipment (e.g., haul trucks and front-end loaders), and loose or... aspects of an assigned task in paragraph (b)(4) of this section, if hazard recognition training specific...

  8. Hydrographic Service Royal Australian Navy

    DTIC Science & Technology

    1993-01-01

    relocation of the Hydrographic Office to Wollongong, NSW; "* A marked decline in chart production caused by workforce changes as a result of the impending...amalgamate the plans of Clinton Coal Loader Wharf and Auckland Point to Barney Point in one plan at scale 1:10 000. The chart now provides continuous

  9. Index to Benet Weapons Laboratory (LCWSL) Technical Reports - 1979

    DTIC Science & Technology

    1980-09-01

    Breech Machanisms Breech Seals Breeches, Cannon Cadmium Sulfide Calibration Cannon Auto Loader Cannon Tube Chemical Vapor Deposition REPORT NUMBER...PAGE(Trh«n Data SntaracQ (Con1t from block #20) Three basic specimen geometries were studied: 1. A symetric specimen; 2. A specimen with +5% wall

  10. Federal Workplace Literacy Project. Internal Evaluation Report.

    ERIC Educational Resources Information Center

    Matuszak, David J.

    This report describes the following components of the Nestle Workplace Literacy Project: six job task analyses, curricula for six workplace basic skills training programs, delivery of courses using these curricula, and evaluation of the process. These six job categories were targeted for training: forklift loader/checker, BB's processing systems…

  11. Standard Electronic Format Specification for Tank Characterization Data Loader Version 3.5

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

    ADAMS, M.R.

    2001-01-31

    The purpose of this document is to describe the standard electronic format for data files that will be sent for entry into the Tank Characterization Database (TCD). There are 2 different file types needed for each data load: (1) Analytical Results and (2) Sample Descriptions.

  12. Low-Capital Systems for Thinning Pine Plantations

    Treesearch

    John Wilhoit; Qingyue Ling; Robert Rummer

    1999-01-01

    Highly mechanized systems utilizing rubber-tired skidders, feller-bunchers, and knuckleboom loaders are the predominant type of timber harvesting operation in the southern United States. These systems, which handle the wood in tree-length form, are highly productive and very efficient, especially for large tracts of timber. Thinnings constitute an increasing proportion...

  13. 29 CFR 1926.1400 - Scope.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... loaders. This machinery is also excluded when used with chains, slings or other rigging to lift suspended... that deliver material to a construction site when the crane is used to transfer building supply sheet goods or building supply packaged materials from the truck crane onto a structure, using a fork/cradle...

  14. 77 FR 38087 - Agency Information Collection Activities; Submission for OMB Review; Comment Request...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-26

    ... protective equipment to protect miners from harmful levels of noise that can result in hearing loss. However..., relies on drills, crushers, compressors, conveyors, trucks, loaders, and other heavy-duty equipment for the excavation, haulage, and processing of material. This equipment creates high sound levels...

  15. 77 FR 16865 - Proposed Extension of Existing Information Collection; Occupational Noise Exposure

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-22

    ..., administrative controls, and personal protective equipment to protect miners from harmful levels of noise that..., compressors, conveyors, trucks, loaders, and other heavy-duty equipment for the excavation, haulage, and processing of material. This equipment creates high sound levels, exposing machine operators as well as...

  16. RNA polymerase gate loop guides the nontemplate DNA strand in transcription complexes.

    PubMed

    NandyMazumdar, Monali; Nedialkov, Yuri; Svetlov, Dmitri; Sevostyanova, Anastasia; Belogurov, Georgiy A; Artsimovitch, Irina

    2016-12-27

    Upon RNA polymerase (RNAP) binding to a promoter, the σ factor initiates DNA strand separation and captures the melted nontemplate DNA, whereas the core enzyme establishes interactions with the duplex DNA in front of the active site that stabilize initiation complexes and persist throughout elongation. Among many core RNAP elements that participate in these interactions, the β' clamp domain plays the most prominent role. In this work, we investigate the role of the β gate loop, a conserved and essential structural element that lies across the DNA channel from the clamp, in transcription regulation. The gate loop was proposed to control DNA loading during initiation and to interact with NusG-like proteins to lock RNAP in a closed, processive state during elongation. We show that the removal of the gate loop has large effects on promoter complexes, trapping an unstable intermediate in which the RNAP contacts with the nontemplate strand discriminator region and the downstream duplex DNA are not yet fully established. We find that although RNAP lacking the gate loop displays moderate defects in pausing, transcript cleavage, and termination, it is fully responsive to the transcription elongation factor NusG. Together with the structural data, our results support a model in which the gate loop, acting in concert with initiation or elongation factors, guides the nontemplate DNA in transcription complexes, thereby modulating their regulatory properties.

  17. Imaging viscoelastic properties of live cells by AFM: power-law rheology on the nanoscale.

    PubMed

    Hecht, Fabian M; Rheinlaender, Johannes; Schierbaum, Nicolas; Goldmann, Wolfgang H; Fabry, Ben; Schäffer, Tilman E

    2015-06-21

    We developed force clamp force mapping (FCFM), an atomic force microscopy (AFM) technique for measuring the viscoelastic creep behavior of live cells with sub-micrometer spatial resolution. FCFM combines force-distance curves with an added force clamp phase during tip-sample contact. From the creep behavior measured during the force clamp phase, quantitative viscoelastic sample properties are extracted. We validate FCFM on soft polyacrylamide gels. We find that the creep behavior of living cells conforms to a power-law material model. By recording short (50-60 ms) force clamp measurements in rapid succession, we generate, for the first time, two-dimensional maps of power-law exponent and modulus scaling parameter. Although these maps reveal large spatial variations of both parameters across the cell surface, we obtain robust mean values from the several hundreds of measurements performed on each cell. Measurements on mouse embryonic fibroblasts show that the mean power-law exponents and the mean modulus scaling parameters differ greatly among individual cells, but both parameters are highly correlated: stiffer cells consistently show a smaller power-law exponent. This correlation allows us to distinguish between wild-type cells and cells that lack vinculin, a dominant protein of the focal adhesion complex, even though the mean values of viscoelastic properties between wildtype and knockout cells did not differ significantly. Therefore, FCFM spatially resolves viscoelastic sample properties and can uncover subtle mechanical signatures of proteins in living cells.

  18. Structural insight into recruitment of translesion DNA polymerase Dpo4 to sliding clamp PCNA

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

    Xing, G.; Kirouac, K.; Shin, Y.J.

    2009-09-16

    DNA polymerases are co-ordinated by sliding clamps (PCNA/{beta}-clamp) in translesion synthesis. It is unclear how these enzymes assemble on PCNA with geometric and functional compatibility. We report the crystal structure of a full-length Y-family polymerase, Dpo4, in complex with heterodimeric PCNA1-PCNA2 at 2.05 {angstrom} resolution. Dpo4 exhibits an extended conformation that differs from the Dpo4 structures in apo- or DNA-bound form. Two hinges have been identified in Dpo4, which render the multidomain polymerase flexible conformations and orientations relative to PCNA. Dpo4 binds specifically to PCNA1 on the conserved ligand binding site. The C-terminal peptide of Dpo4 becomes structured with amore » 3{sub 10} helix and dominates the specific binding. The Y-family polymerase also contacts PCNA1 with its finger, thumb and little finger domains, which are conformation-dependent protein-protein interactions that diversify the binding mode of Dpo4 on PCNA. The structure reveals a molecular model in which substrate/partner binding-coupled multiple conformations of a Y-family polymerase facilitate its recruitment and co-ordination on the sliding clamp. The conformational flexibility would turn the error-prone Y-family polymerase off when more efficient high-fidelity DNA polymerases work on undamaged DNA and turn it onto DNA templates to perform translesion synthesis when replication forks are stalled by DNA lesions.« less

  19. Impact of Renal Hilar Control on Outcomes of Robotic Partial Nephrectomy: Systematic Review and Cumulative Meta-analysis.

    PubMed

    Cacciamani, Giovanni E; Medina, Luis G; Gill, Tania S; Mendelsohn, Alec; Husain, Fatima; Bhardwaj, Lokesh; Artibani, Walter; Sotelo, Renè; Gill, Inderbir S

    2018-02-05

    During robotic partial nephrectomy (RPN), various techniques of hilar control have been described, including on-clamp, early unclamping, selective/super-selective clamping, and completely-unclamped RPN. To evaluate the impact of various hilar control techniques on perioperative, functional, and oncological outcomes of RPN for tumors. We conducted a systematic literature review and meta-analysis of all comparative studies on various hilar control techniques during RPN using PubMed, Scopus, and Web of Science according to the Preferred Reporting Items for Systematic Review and Meta-analysis statement, and Methods and Guide for Effectiveness and Comparative Effectiveness Review of the Agency for Healthcare Research and Quality. Cumulative meta-analysis of comparative studies was conducted using Review Manager 5.3. Of 987 RPN publications in the literature, 19 qualified for this analysis. Comparison of off-clamp versus on-clamp RPN (n=9), selective clamping versus on-clamp RPN (n=3), super selective clamping versus on-clamp RPN (n=5), and early unclamped versus on-clamp (n=3) were reported. Patients undergoing RPN using off-clamp, selective/super selective, or early unclamp techniques had higher estimated blood loss compared with on-clamp RPN (weight mean difference [WMD]: 47.83, p=0.000, WMD: 41.06, p=0.02, and WMD: 37.50, p=0.47); however, this did not seem clinically relevant, since transfusion rates were similar (odds ratio [OR]: 0.98, p=0.95, OR: 0.72, p=0.7, and OR: 1.36, p=0.33, respectively). All groups appeared similar with regards to hospital stay, transfusions, overall and major complications, and positive cancer margin rates. Short- and long-term renal functional outcomes appeared superior in the off-clamp and super selective clamp groups compared with the on-clamp RPN cohort. Off-clamp, selective/super selective clamp, and early unclamp hilar control techniques are safe and feasible approaches for RPN surgery, with similar perioperative and oncological outcomes compared with on-clamp RPN. Minimizing global renal ischemia may provide superior renal function preservation. However, higher quality data are necessary for definitive conclusions in this regard. The objective of partial nephrectomy is to treat the cancer while maximizing renal function preservation. Clamping the main vessels is done primarily to reduce the blood loss during partial nephrectomy; however, vascular clamping can compromise kidney function. In order to avoid clamping, various techniques have been described. Our analysis showed that techniques that avoid main renal artery clamping during RPN are associated with better renal function preservation, yet deliver non-inferior perioperative and oncological outcomes as compared with robotic partial nephrectomy procedures that clamp the main vessels. Copyright © 2018 European Association of Urology. Published by Elsevier B.V. All rights reserved.

  20. Can the learning curve of totally endoscopic robotic mitral valve repair be short-circuited?

    PubMed

    Yaffee, David W; Loulmet, Didier F; Kelly, Lauren A; Ward, Alison F; Ursomanno, Patricia A; Rabinovich, Annette E; Neuburger, Peter J; Krishnan, Sandeep; Hill, Frederick T; Grossi, Eugene A

    2014-01-01

    A concern with the initiation of totally endoscopic robotic mitral valve repair (TERMR) programs has been the risk for the learning curve. To minimize this risk, we initiated a TERMR program with a defined team and structured learning approach before clinical implementation. A dedicated team (two surgeons, one cardiac anesthesiologist, one perfusionist, and two nurses) was trained with clinical scenarios, simulations, wet laboratories, and "expert" observation for 3 months. This team then performed a series of TERMRs of varying complexity. Thirty-two isolated TERMRs were performed during the first programmatic year. All operations included mitral valve repair, left atrial appendage exclusion, and annuloplasty device implantation. Additional procedures included leaflet resection, neochordae insertion, atrial ablation, and papillary muscle shortening. Longer clamp times were associated with number of neochordae (P < 0.01), papillary muscle procedures (P < 0.01), and leaflet resection (P = 0.06). Sequential case number had no impact on cross-clamp time (P = 0.3). Analysis of nonclamp time demonstrated a 71.3% learning percentage (P < 0.01; ie, 28.7% reduction in nonclamp time with each doubling of case number). There were no hospital deaths or incidences of stroke, myocardial infarction, unplanned reoperation, respiratory failure, or renal failure. Median length of stay was 4 days. All patients were discharged home. Totally endoscopic robotic mitral valve repair can be safely performed after a pretraining regimen with emphasis on experts' current practice and team training. After a pretraining regimen, cross-clamp times were not subject to learning curve phenomena but were dependent on procedural complexity. Nonclamp times were associated with a short learning curve.

  1. Modulation by clamping: Kv4 and KChIP interactions.

    PubMed

    Wang, Kewei

    2008-10-01

    The rapidly inactivating (A-type) potassium channels regulate membrane excitability that defines the fundamental mechanism of neuronal functions such as pain signaling. Cytosolic Kv channel-interacting proteins KChIPs that belong to neuronal calcium sensor (NCS) family of calcium binding EF-hand proteins co-assemble with Kv4 (Shal) alpha subunits to form a native complex that encodes major components of neuronal somatodendritic A-type K+ current, I(SA), in neurons and transient outward current, I(TO), in cardiac myocytes. The specific binding of auxiliary KChIPs to the Kv4 N-terminus results in modulation of gating properties, surface expression and subunit assembly of Kv4 channels. Here, I attempt to emphasize the interaction between KChIPs and Kv4 based on recent progress made in understanding the structure complex in which a single KChIP1 molecule laterally clamps two neighboring Kv4.3 N-termini in a 4:4 manner. Greater insights into molecular mechanism between KChIPs and Kv4 interaction may provide therapeutic potentials of designing compounds aimed at disrupting the protein-protein interaction for treatment of membrane excitability-related disorders.

  2. Clamp usable as jig and lifting clamp

    DOEpatents

    Tsuyama, Yoshizo

    1976-01-01

    There is provided a clamp which is well suited for use as a lifting clamp for lifting and moving materials of assembly in a shipyard, etc. and as a pulling jig in welding and other operations. The clamp comprises a clamp body including a shackle for engagement with a pulling device and a slot for receiving an article, and a pair of jaws provided on the leg portions of the clamp body on the opposite sides of the slot to grip the article in the slot, one of said jaws consisting of a screw rod and the other jaw consisting of a swivel jaw with a spherical surface, whereby when the article clamped in the slot by the pair of jaws tends to slide in any direction with respect to the clamp body, the article is more positively gripped by the pair of jaws.

  3. Cable clamp bolt fixture facilitates assembly in close quarters

    NASA Technical Reports Server (NTRS)

    Sunderland, G. H.

    1967-01-01

    Cable clamp bolt holding fixture facilitates forming of electrical cable runs in limited equipment space. The fixture engages the threads of the short clamp bolt through the clamp and maintains tension against clamp tendency to open while the operator installs the nut without difficulty.

  4. Split-tapered joint clamping device

    DOEpatents

    Olsen, Max J.; Schwartz, Jr., John F.

    1988-01-01

    This invention relates to a clamping device for removably attaching a tool element to a bracket element wherein a bracket element is disposed in a groove in the tool and a clamping member is disposed in said groove and in engagement with a clamping face of the bracket and a wall of the groove and with the clamping member having pivot means engaging the bracket and about which the clamping member rotates.

  5. Research on mechanical and sensoric set-up for high strain rate testing of high performance fibers

    NASA Astrophysics Data System (ADS)

    Unger, R.; Schegner, P.; Nocke, A.; Cherif, C.

    2017-10-01

    Within this research project, the tensile behavior of high performance fibers, such as carbon fibers, is investigated under high velocity loads. This contribution (paper) focuses on the clamp set-up of two testing machines. Based on a kinematic model, weight optimized clamps are designed and evaluated. By analyzing the complex dynamic behavior of conventional high velocity testing machines, it has been shown that the impact typically exhibits an elastic characteristic. This leads to barely predictable breaking speeds and will not work at higher speeds when acceleration force exceeds material specifications. Therefore, a plastic impact behavior has to be achieved, even at lower testing speeds. This type of impact behavior at lower speeds can be realized by means of some minor test set-up adaptions.

  6. Effect of complete hilar versus only renal artery clamping on renal histomorphology following ischemia/reperfusion injury in an experimental model.

    PubMed

    Umul, M; Cal, A C; Turna, B; Oktem, G; Aydın, H H

    2016-01-01

    To evaluate the effect of temporary complete hilar versus only renal artery clamping with different duration of warm ischemia on renal functions, and possibly identify a "safe" clamping type and duration of renal ischemia. Fifty male rabbits have been incorporated to study. Rabbits were subjected to ischemia/reperfusion injury by temporary vascular clamping. Reagents were randomized to 3 experimental groups (only renal artery clamping, complete hilar clamping, sham surgery) and sub-groups were determined according to different clamping times (30 and 60 minutes). Median laparotomy and left renal hilus dissection were performed to sham group. Only artery or complete hilar clamping was performed for 30 or 60 minutes by microvascular bulldog clamps to other reagents. Rabbits were sacrificed 10 days after primary surgery and left nephrectomy performed. Nephrectomy materials were evaluated for the level of nitric-oxide synthase (NOS) immunoreactivity, malondialdehyde (MDA) level and superoxide dismutase (SOD) activity and an electron microscopic examination was performed. NOS immunoreactivity was correlated with the temporary clamping time. We also observed that complete hilar vascular clamping entails an increase on NOS immunoreactivity. MDA levels were similar for all experimental surgery groups (p = 0.42). The SOD activity was decreased among all subgroups compared with sham surgery. But the significant decrease occurred in 30 minutes only artery and 30 minutes complete hilar clamping groups in proportion to sham surgery (p = 0.026 and p = 0.019, respectively). This current study suggested that only renal artery clamping under 30 minutes is more appropriate during renal surgical procedures requiring temporary vascular clamping.

  7. Lifting clamp positively grips structural shapes

    NASA Technical Reports Server (NTRS)

    Reinhardt, E. C.

    1966-01-01

    Welded steel clamps securely grip structural shapes of various sizes for crane operations. The clamp has adjustable clamping jaws and screw-operated internal v-jaws and provides greater safety than hoisting slings presently used. The structural member can be rotated in any manner, angle, or direction without being released by the clamp.

  8. The β2 clamp in the Mycobacterium tuberculosis DNA polymerase III αβ2ε replicase promotes polymerization and reduces exonuclease activity

    PubMed Central

    Gu, Shoujin; Li, Wenjuan; Zhang, Hongtai; Fleming, Joy; Yang, Weiqiang; Wang, Shihua; Wei, Wenjing; Zhou, Jie; Zhu, Guofeng; Deng, Jiaoyu; Hou, Jian; Zhou, Ying; Lin, Shiqiang; Zhang, Xian-En; Bi, Lijun

    2016-01-01

    DNA polymerase III (DNA pol III) is a multi-subunit replication machine responsible for the accurate and rapid replication of bacterial genomes, however, how it functions in Mycobacterium tuberculosis (Mtb) requires further investigation. We have reconstituted the leading-strand replication process of the Mtb DNA pol III holoenzyme in vitro, and investigated the physical and functional relationships between its key components. We verify the presence of an αβ2ε polymerase-clamp-exonuclease replicase complex by biochemical methods and protein-protein interaction assays in vitro and in vivo and confirm that, in addition to the polymerase activity of its α subunit, Mtb DNA pol III has two potential proofreading subunits; the α and ε subunits. During DNA replication, the presence of the β2 clamp strongly promotes the polymerization of the αβ2ε replicase and reduces its exonuclease activity. Our work provides a foundation for further research on the mechanism by which the replication machinery switches between replication and proofreading and provides an experimental platform for the selection of antimicrobials targeting DNA replication in Mtb. PMID:26822057

  9. NUMERICAL SIMULATION OF NANOINDENTATION AND PATCH CLAMP EXPERIMENTS ON MECHANOSENSITIVE CHANNELS OF LARGE CONDUCTANCE IN ESCHERICHIA COLI

    PubMed Central

    Tang, Yuye; Chen, Xi; Yoo, Jejoong; Yethiraj, Arun; Cui, Qiang

    2010-01-01

    A hierarchical simulation framework that integrates information from all-atom simulations into a finite element model at the continuum level is established to study the mechanical response of a mechanosensitive channel of large conductance (MscL) in bacteria Escherichia Coli (E.coli) embedded in a vesicle formed by the dipalmitoylphosphatidycholine (DPPC) lipid bilayer. Sufficient structural details of the protein are built into the continuum model, with key parameters and material properties derived from molecular mechanics simulations. The multi-scale framework is used to analyze the gating of MscL when the lipid vesicle is subjective to nanoindentation and patch clamp experiments, and the detailed structural transitions of the protein are obtained explicitly as a function of external load; it is currently impossible to derive such information based solely on all-atom simulations. The gating pathways of E.coli-MscL qualitatively agree with results from previous patch clamp experiments. The gating mechanisms under complex indentation-induced deformation are also predicted. This versatile hierarchical multi-scale framework may be further extended to study the mechanical behaviors of cells and biomolecules, as well as to guide and stimulate biomechanics experiments. PMID:21874098

  10. Digital force-feedback for protein unfolding experiments using atomic force microscopy

    NASA Astrophysics Data System (ADS)

    Bippes, Christian A.; Janovjak, Harald; Kedrov, Alexej; Muller, Daniel J.

    2007-01-01

    Since its invention in the 1990s single-molecule force spectroscopy has been increasingly applied to study protein (un-)folding, cell adhesion, and ligand-receptor interactions. In most force spectroscopy studies, the cantilever of an atomic force microscope (AFM) is separated from a surface at a constant velocity, thus applying an increasing force to folded bio-molecules or bio-molecular bonds. Recently, Fernandez and co-workers introduced the so-called force-clamp technique. Single proteins were subjected to a defined constant force allowing their life times and life time distributions to be directly measured. Up to now, the force-clamping was performed by analogue PID controllers, which require complex additional hardware and might make it difficult to combine the force-feedback with other modes such as constant velocity. These points may be limiting the applicability and versatility of this technique. Here we present a simple, fast, and all-digital (software-based) PID controller that yields response times of a few milliseconds in combination with a commercial AFM. We demonstrate the performance of our feedback loop by force-clamp unfolding of single Ig27 domains of titin and the membrane proteins bacteriorhodopsin (BR) and the sodium/proton antiporter NhaA.

  11. 30 CFR Appendix I to Subpart M of... - National Consensus Standards

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... loaders and bulldozers Society of Automotive Engineers (SAE) minimum performance criteria for falling object protective structures (FOPS) SAE J231—January, 1981. Fork-lift trucks American National Standards Institute (ANSI) safety standard for low lift and high lift trucks, B 56.1, section 7.27—1983; or, American...

  12. 30 CFR Appendix I to Subpart M of... - National Consensus Standards

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... loaders and bulldozers Society of Automotive Engineers (SAE) minimum performance criteria for falling object protective structures (FOPS) SAE J231—January, 1981. Fork-lift trucks American National Standards Institute (ANSI) safety standard for low lift and high lift trucks, B 56.1, section 7.27—1983; or, American...

  13. 30 CFR Appendix I to Subpart M of... - National Consensus Standards

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... loaders and bulldozers Society of Automotive Engineers (SAE) minimum performance criteria for falling object protective structures (FOPS) SAE J231—January, 1981. Fork-lift trucks American National Standards Institute (ANSI) safety standard for low lift and high lift trucks, B 56.1, section 7.27—1983; or, American...

  14. 30 CFR Appendix I to Subpart M of... - National Consensus Standards

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... loaders and bulldozers Society of Automotive Engineers (SAE) minimum performance criteria for falling object protective structures (FOPS) SAE J231—January, 1981. Fork-lift trucks American National Standards Institute (ANSI) safety standard for low lift and high lift trucks, B 56.1, section 7.27—1983; or American...

  15. 30 CFR Appendix I to Subpart M of... - National Consensus Standards

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... loaders and bulldozers Society of Automotive Engineers (SAE) minimum performance criteria for falling object protective structures (FOPS) SAE J231—January, 1981. Fork-lift trucks American National Standards Institute (ANSI) safety standard for low lift and high lift trucks, B 56.1, section 7.27—1983; or American...

  16. 30 CFR Appendix I to Subpart M of... - National Consensus Standards

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... loaders and bulldozers Society of Automotive Engineers (SAE) minimum performance criteria for falling object protective structures (FOPS) SAE J231—January, 1981. Fork-lift trucks American National Standards Institute (ANSI) safety standard for low lift and high lift trucks, B 56.1, section 7.27—1983; or American...

  17. 30 CFR 46.6 - Newly hired experienced miner training.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...) Instruction on the recognition and avoidance of electrical hazards and other hazards present at the mine, such as traffic patterns and control, mobile equipment (e.g., haul trucks and front-end loaders), and... health and safety aspects of an assigned task in paragraph (b)(4) of this section, if hazard recognition...

  18. 30 CFR 46.6 - Newly hired experienced miner training.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...) Instruction on the recognition and avoidance of electrical hazards and other hazards present at the mine, such as traffic patterns and control, mobile equipment (e.g., haul trucks and front-end loaders), and... health and safety aspects of an assigned task in paragraph (b)(4) of this section, if hazard recognition...

  19. 76 FR 36548 - Circulatory System Devices Panel of the Medical Devices Advisory Committee; Notice of Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-22

    ...] Circulatory System Devices Panel of the Medical Devices Advisory Committee; Notice of Meeting AGENCY: Food and... of Committee: Circulatory System Devices Panel of the Medical Devices Advisory Committee. General... also comes with a sheath, introducer, loader, dilator, balloon (used to pre-dilate the native annulus...

  20. 77 FR 25183 - Circulatory System Devices Panel of the Medical Devices Advisory Committee; Notice of Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-27

    ...] Circulatory System Devices Panel of the Medical Devices Advisory Committee; Notice of Meeting AGENCY: Food and... of Committee: Circulatory System Devices Panel of the Medical Devices Advisory Committee. General..., introducer, loader, dilator, balloon (used to pre-dilate the native annulus) and a crimper. FDA intends to...

  1. 40 CFR Table 9 to Subpart Wwww of... - Initial Compliance With Work Practice Standards

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... compression/injection molding uncover, unwrap or expose only one charge per mold cycle per compression/injection molding machine. For machines with multiple molds, one charge means sufficient material to fill... cycle per compression/injection molding machine, or prior to the loader, hoppers are closed except when...

  2. --No Title--

    Science.gov Websites

    background-color: #e5e5e5; } h3 { line-height: 20px; padding-bottom: 10px; } #appheader { margin ; position: relative; text-align: center; font-size: 16pt; /*font-weight: bold;*/ /*color: rgb(165,180,212 );*/ color: rgb(62, 96, 163); background: url('../images/ajax-loader.gif') 50% 50% no-repeat; } .container

  3. The Impact of Wireless Technology on Loading Trucks at an Auto Parts Distribution Center

    ERIC Educational Resources Information Center

    Goomas, David T.

    2012-01-01

    An intervention was introduced for truck loaders that used wireless vehicle mount computers that included auditory, visual feedback, and immediate data delivery. The implementation reliably reduced pallets from being loaded out of sequence for all outbound trucks in multistop routes. The role of the organizational behavior management (OBM)…

  4. Gamma-ray exposure hazard due to stowage of M-774 APFSDS (Armour-piercing fin-stabilized discarding-sabot) rounds in a Leopard C-1 main battle tank. Technical note

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

    Robitaille, H.A.

    1983-09-01

    The M-744 APFSDS anti-tank round contains a depleted-uranium penetrator of 3.4 kgram mass. Since depleted uranium is naturally radioactive, the storage of a substantial number of these rounds inside a Leopard C-1 main battle tank creates a gamma radiation field within the vehicle. Gamma-ray exposure rates have been measured using a sodium-iodide spectrometer at each of the four crew-member locations and for two turret orientations, with 59 rounds stowed in the vehicle. In all cases the measured gamma-ray dose rates were less than a maximum of 0.17 millirad per hour observed at the loader's position. Assuming the loader spent anmore » entire week (168 hours) at his station, his integrated dose would amount to 29 mRad - approximately a factor of 4 lower than the maximum currently allowed by Canadian Forces regulations. It is therefore concluded that the M774 round does not represent a significant gamma radiation hazard to Leopard C1 crewmembers.« less

  5. Whole-Cell Electrical Activity Under Direct Mechanical Stimulus by AFM Cantilever Using Planar Patch Clamp Chip Approach

    PubMed Central

    Upadhye, Kalpesh V.; Candiello, Joseph E.; Davidson, Lance A.; Lin, Hai

    2011-01-01

    Patch clamp is a powerful tool for studying the properties of ion-channels and cellular membrane. In recent years, planar patch clamp chips have been fabricated from various materials including glass, quartz, silicon, silicon nitride, polydimethyl-siloxane (PDMS), and silicon dioxide. Planar patch clamps have made automation of patch clamp recordings possible. However, most planar patch clamp chips have limitations when used in combination with other techniques. Furthermore, the fabrication methods used are often expensive and require specialized equipments. An improved design as well as fabrication and characterization of a silicon-based planar patch clamp chip are described in this report. Fabrication involves true batch fabrication processes that can be performed in most common microfabrication facilities using well established MEMS techniques. Our planar patch clamp chips can form giga-ohm seals with the cell plasma membrane with success rate comparable to existing patch clamp techniques. The chip permits whole-cell voltage clamp recordings on variety of cell types including Chinese Hamster Ovary (CHO) cells and pheochromocytoma (PC12) cells, for times longer than most available patch clamp chips. When combined with a custom microfluidics chamber, we demonstrate that it is possible to perfuse the extra-cellular as well as intra-cellular buffers. The chamber design allows integration of planar patch clamp with atomic force microscope (AFM). Using our planar patch clamp chip and microfluidics chamber, we have recorded whole-cell mechanosensitive (MS) currents produced by directly stimulating human keratinocyte (HaCaT) cells using an AFM cantilever. Our results reveal the spatial distribution of MS ion channels and temporal details of the responses from MS channels. The results show that planar patch clamp chips have great potential for multi-parametric high throughput studies of ion channel proteins. PMID:22174731

  6. Measuring beta-cell function relative to insulin sensitivity in youth: Does the hyperglycemic clamp suffice?

    USDA-ARS?s Scientific Manuscript database

    To compare beta-cell function relative to insulin sensitivity, disposition index (DI), calculated from two clamps (2cDI, insulin sensitivity from the hyperinsulinemic-euglycemic clamp and first-phase insulin from the hyperglycemic clamp) with the DI calculated from the hyperglycemic clamp alone (hcD...

  7. 21 CFR 882.4460 - Neurosurgical head holder (skull clamp).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Neurosurgical head holder (skull clamp). 882.4460... holder (skull clamp). (a) Identification. A neurosurgical head holder (skull clamp) is a device used to clamp the patient's skull to hold head and neck in a particular position during surgical procedures. (b...

  8. Close relationship of tissue plasminogen activator-plasminogen activator inhibitor-1 complex with multiple organ dysfunction syndrome investigated by means of the artificial pancreas

    PubMed Central

    Hoshino, Masami; Haraguchi, Yoshikura; Hirasawa, Hiroyuki; Sakai, Motohiro; Saegusa, Hiroshi; Hayashi, Kazushiro; Horita, Naoki; Ohsawa, Hiroyuki

    2001-01-01

    Background: Glucose tolerance (GT) has not been taken into consideration in investigations concerning relationships between coagulopathy and multiple organ dysfunction syndrome (MODS), and endothelial cell activation/endothelial cell injury (ECA/ECI) in septic patients, although coagulopathy is known to be influenced by blood glucose level. We investigated those relationships under strict blood glucose control and evaluation of GT with the glucose clamp method by means of the artificial pancreas in nine septic patients with glucose intolerance. The relationships between GT and blood stress related hormone levels (SRH) were also investigated. Methods: The amount of metabolized glucose (M value), as the parameter of GT, was measured by the euglycemic hyperinsulinemic glucose clamp method, in which the blood glucose level was clamped at 80 mg/dl under a continuous insulin infusion rate of 1.12 mU/kg per min, using the artificial pancreas, STG-22. Multiple organ failure (MOF) score was calculated using the MOF criteria of Japanese Association for Critical Care Medicine. Regarding coagulopathy, the following parameters were used: disseminated intravascular coagulation (DIC) score (calculated from the DIC criteria of the Ministry of Health and Welfare of Japan) and the parameters used for calculating DIC score, protein-C, protein-S, plasminogen, antithrombin III (AT-III), plasminogen activator inhibitor-1 (PAI-1), and tissue plasminogen activator-PAI-1 (tPA-PAI-1) complex. Thrombomodulin (TM) was measured as the indicator of ECI. Results: There were no significant correlations between M value and SRH, parameters indicating coagulopathy and the MOF score. The MOF score and blood TM levels were positively correlated with DIC score, thrombin-AT-III complex and tPA-PAI-1 complex, and negatively correlated with blood platelet count. Conclusions: GT was not significantly related to SRH, coagulopathy and MODS under strict blood glucose control. Hypercoagulability was closely related to MODS and ECI. Among the parameters indicating coagulopathy, tPA-PAI-1 complex, which is considered to originate from ECA, seemed to be a sensitive parameter of MODS and ECI, and might be a predictive marker of MODS. The treatment for reducing hypercoagulability and ECA/ECI were thought to be justified as one of the therapies for acutely ill septic patients. PMID:11299067

  9. Microchip amplifier for in vitro, in vivo, and automated whole cell patch-clamp recording

    PubMed Central

    Kolb, Ilya; Kodandaramaiah, Suhasa B.; Chubykin, Alexander A.; Yang, Aimei; Bear, Mark F.; Boyden, Edward S.; Forest, Craig R.

    2014-01-01

    Patch clamping is a gold-standard electrophysiology technique that has the temporal resolution and signal-to-noise ratio capable of reporting single ion channel currents, as well as electrical activity of excitable single cells. Despite its usefulness and decades of development, the amplifiers required for patch clamping are expensive and bulky. This has limited the scalability and throughput of patch clamping for single-ion channel and single-cell analyses. In this work, we have developed a custom patch-clamp amplifier microchip that can be fabricated using standard commercial silicon processes capable of performing both voltage- and current-clamp measurements. A key innovation is the use of nonlinear feedback elements in the voltage-clamp amplifier circuit to convert measured currents into logarithmically encoded voltages, thereby eliminating the need for large high-valued resistors, a factor that has limited previous attempts at integration. Benchtop characterization of the chip shows low levels of current noise [1.1 pA root mean square (rms) over 5 kHz] during voltage-clamp measurements and low levels of voltage noise (8.2 μV rms over 10 kHz) during current-clamp measurements. We demonstrate the ability of the chip to perform both current- and voltage-clamp measurement in vitro in HEK293FT cells and cultured neurons. We also demonstrate its ability to perform in vivo recordings as part of a robotic patch-clamping system. The performance of the patch-clamp amplifier microchip compares favorably with much larger commercial instrumentation, enabling benchtop commoditization, miniaturization, and scalable patch-clamp instrumentation. PMID:25429119

  10. Quantification of atmospheric emissions and energy metrics from simulated clamp kiln technology in the clay brick industry.

    PubMed

    Akinshipe, Oladapo; Kornelius, Gerrit

    2018-05-01

    The quantification of atmospheric emissions from clamp kilns in the clay brick industry has met with limited success globally. The complex configuration of clamp kilns using coal or other carbonaceous fuels and uncertainties regarding kiln combustion conditions, has proven to be a hurdle in measurement of emissions and standardization of process and energy metrics. To enable quantification of these metrics, a model kiln was designed to simulate operating conditions and configuration similar to transverse slice of a typical full-scale clamp kiln, but with lower capacity (20,000 to 35,000 bricks per cycle). Hourly measurements of flue gas at extraction duct were recorded for thirteen firing cycles obtained from various source factories, each lasting 8-14 days, for SO 2 , NO 2 , NO, PM, CO and CO 2 emissions in the extraction stack. A statistical mean efficiency for model kiln emissions capturing and channelling capacity was calculated from sulfur mass balance results of batches that lie within 95% confidence interval of assumed true mean (100%) to give 84.2%. Final emission factors (mean ± standard deviation) were quantified as 22.5 ± 18.8 g/brick for CO, 0.14 ± 0.1 g/brick for NO, 0.0 g/brick for NO 2 , 0.14 ± 0.1 g/brick for NO x , 1.07 ± 0.7 g/brick for SO 2 , 378 ± 223 g/brick for CO 2 ; 0.96 ± 0.5 g/brick for PM 10 ; as well as 1.53 g/brick for hydrocarbons. Energy analysis indicate that a significant reduction of 0.9 MJ/kg (36%) in energy use could be achieved by clamp kiln operators, thereby reducing input costs, and significantly reducing atmospheric emissions. Copyright © 2018 Elsevier Ltd. All rights reserved.

  11. Photovoltaic panel clamp

    DOEpatents

    Mittan, Margaret Birmingham [Oakland, CA; Miros, Robert H. J. [Fairfax, CA; Brown, Malcolm P [San Francisco, CA; Stancel, Robert [Loss Altos Hills, CA

    2012-06-05

    A photovoltaic panel clamp includes an upper and lower section. The interface between the assembled clamp halves and the module edge is filled by a flexible gasket material, such as EPDM rubber. The gasket preferably has small, finger like protrusions that allow for easy insertion onto the module edge while being reversed makes it more difficult to remove them from the module once installed. The clamp includes mounting posts or an integral axle to engage a bracket. The clamp also may include a locking tongue to secure the clamp to a bracket.

  12. Photovoltaic panel clamp

    DOEpatents

    Brown, Malcolm P.; Mittan, Margaret Birmingham; Miros, Robert H. J.; Stancel, Robert

    2013-03-19

    A photovoltaic panel clamp includes an upper and lower section. The interface between the assembled clamp halves and the module edge is filled by a flexible gasket material, such as EPDM rubber. The gasket preferably has small, finger like protrusions that allow for easy insertion onto the module edge while being reversed makes it more difficult to remove them from the module once installed. The clamp includes mounting posts or an integral axle to engage a bracket. The clamp also may include a locking tongue to secure the clamp to a bracket.

  13. Near-infrared imaging of face transplants: are both pedicles necessary?

    PubMed

    Nguyen, John T; Ashitate, Yoshitomo; Venugopal, Vivek; Neacsu, Florin; Kettenring, Frank; Frangioni, John V; Gioux, Sylvain; Lee, Bernard T

    2013-09-01

    Facial transplantation is a complex procedure that corrects severe facial defects due to traumas, burns, and congenital disorders. Although face transplantation has been successfully performed clinically, potential risks include tissue ischemia and necrosis. The vascular supply is typically based on the bilateral neck vessels. As it remains unclear whether perfusion can be based off a single pedicle, this study was designed to assess perfusion patterns of facial transplant allografts using near-infrared (NIR) fluorescence imaging. Upper facial composite tissue allotransplants were created using both carotid artery and external jugular vein pedicles in Yorkshire pigs. A flap validation model was created in n = 2 pigs and a clamp occlusion model was performed in n = 3 pigs. In the clamp occlusion models, sequential clamping of the vessels was performed to assess perfusion. Animals were injected with indocyanine green and imaged with NIR fluorescence. Quantitative metrics were assessed based on fluorescence intensity. With NIR imaging, arterial perforators emitted fluorescence indicating perfusion along the surface of the skin. Isolated clamping of one vascular pedicle showed successful perfusion across the midline based on NIR fluorescence imaging. This perfusion extended into the facial allograft within 60 s and perfused the entire contralateral side within 5 min. Determination of vascular perfusion is important in microsurgical constructs as complications can lead to flap loss. It is still unclear if facial transplants require both pedicles. This initial pilot study using intraoperative NIR fluorescence imaging suggests that facial flap models can be adequately perfused from a single pedicle. Copyright © 2013 Elsevier Inc. All rights reserved.

  14. Saddle clamp assembly

    NASA Technical Reports Server (NTRS)

    Belrose, Charles R. (Inventor)

    1994-01-01

    A saddle clamp assembly is presented. The assembly is comprised of a hollow cylindrical body centered about a longitudinal axis and being diametrically split into semicircular top and bottom sections. Each section has a pair of connection flanges, at opposite ends, that project radially outward. A pair of bolts are retained on the top section flanges and are threadable into nuts retained on the bottom section flanges. A base member is anchored to a central underside portion of the bottom clamp body section and has a pair of connection tabs positioned beneath the bottom clamp body section connection flanges on opposite sides of the clamp axis. A pair of bolts are retained on the base member connection tabs and are threadable into a pair of nuts retainable on a support structure. The connection tab and connection flanges on each side of the clamp body are axially offset in a manner permitting downward installation/removable tool access to the lower bolts past the connection flanges. An elongated retention tether is used to connect the top clamp body section to the balance of the clamp assembly. This prevents loss of the top clamp body section when it is removed from the bottom clamp body section.

  15. Analysis of failed and nickel-coated 3093 beam clamp components at the East Tennessee Technology Park (ETTP).

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

    Singh, D.; Pappacena, K.; Gaviria, J.

    2010-10-11

    The U.S. Department of Energy and its contractor, Bechtel Jacobs Company (BJC), are undertaking a major effort to clean up the former gaseous diffusion facility (K-25) located in Oak Ridge, TN. The decontamination and decommissioning activities require systematic removal of contaminated equipment and machinery followed by demolition of the buildings. As part of the cleanup activities, a beam clamp, used for horizontal life lines (HLLs) for fall protection, was discovered to be fractured during routine inspection. The beam clamp (yoke and D-ring) was a component in the HLL system purchased from Reliance Industries LLC. Specifically, the U-shaped stainless steel yokemore » of the beam clamp failed in a brittle mode at under less than 10% of the rated design capacity of 14,500 lb. The beam clamp had been in service for approximately 16 months. Bechtel Jacobs approached Argonne National Laboratory to assist in identifying the root cause of the failure of the beam clamp. The objectives of this study were to (1) review the prior reports and documents on the subject, (2) understand the possible failure mechanism(s) that resulted in the failed beam clamp components, (3) recommend approaches to mitigate the failure mechanism(s), and (4) evaluate the modified beam clamp assemblies. Energy dispersive x-ray analysis and chemical analysis of the corrosion products on the failed yoke and white residue on an in-service yoke indicated the presence of zinc, sulfur, and calcium. Analysis of rainwater in the complex, as conducted by BJC, indicated the presence of sulfur and calcium. It was concluded that, as a result of galvanic corrosion, zinc from the galvanized components of the beam clamp assembly (D-ring) migrated to the corroded region in the presence of the rainwater. Under mechanical stress, the corrosion process would have accelerated, resulting in the catastrophic failure of the yoke. As suggested by Bechtel Jacobs personnel, hydrogen embrittlement as a consequence of corrosion was also explored as a failure mechanism. Corroded and failed yoke samples had hydrogen concentrations of 20-60 ppm. However, the hydrogen content reduced to 4-11 ppm (similar to baseline as-received yoke samples) when the corrosion products were polished off. The hydrogen content in the scraped off corrosion product powders was >7000 ppm. These results indicate that hydrogen is primarily present in the corrosion products and not in the underlying steel. Rockwell hardness values on the corroded yoke and D-rings were R{sub c} {approx} 41-46. It was recommended to the beam clamp manufacturer that the beam clamp components be annealed to reduce the hardness values so that they are less susceptible to brittle failure. Upon annealing, hardness values of the beam clamp components reduced to R{sub c} {approx} 25. Several strategies were recommended and put in place to mitigate failure of the beam clamp components: (a) maintain hardness levels of both yokes and D-rings at R{sub c} < 35, (b) coat the yoke and D-rings with a dual coating of nickel (with 10% phosphorus) to delay corrosion and aluminum to prevent galvanic corrosion since it is more anodic to zinc, and (c) optimize coating thicknesses for nickel and aluminum while maintaining the physical integrity of the coatings. Evaluation of the Al- and Ni-coated yoke and D-ring specimens indicated they appear to have met the recommendations. Average hardness values of the dual-coated yokes were R{sub c} {approx} 25-35. Hardness values of dual-coated D-ring were R{sub c} {approx} 32. Measured average coating thicknesses for the aluminum and nickel coatings for yoke samples were 22 {micro}m (0.9 mils) and 80 {micro}m (3 mils), respectively. The D-rings also showed similar coating thicknesses. Microscopic examination showed that the aluminum coating was well bonded to the underlying nickel coating. Some observed damage was believed to be an artifact of the cutting-and-polishing steps during sample preparation for microscopy.« less

  16. 46 CFR 128.420 - Keel cooler installations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...-metallic hose-clamps may be used at machinery connections if— (1) The clamps are of a corrosion-resistant material; (2) The clamps do not depend on spring tension for their holding power; and (3) Two of the clamps...

  17. High throughput ion-channel pharmacology: planar-array-based voltage clamp.

    PubMed

    Kiss, Laszlo; Bennett, Paul B; Uebele, Victor N; Koblan, Kenneth S; Kane, Stefanie A; Neagle, Brad; Schroeder, Kirk

    2003-02-01

    Technological advances often drive major breakthroughs in biology. Examples include PCR, automated DNA sequencing, confocal/single photon microscopy, AFM, and voltage/patch-clamp methods. The patch-clamp method, first described nearly 30 years ago, was a major technical achievement that permitted voltage-clamp analysis (membrane potential control) of ion channels in most cells and revealed a role for channels in unimagined areas. Because of the high information content, voltage clamp is the best way to study ion-channel function; however, throughput is too low for drug screening. Here we describe a novel breakthrough planar-array-based HT patch-clamp technology developed by Essen Instruments capable of voltage-clamping thousands of cells per day. This technology provides greater than two orders of magnitude increase in throughput compared with the traditional voltage-clamp techniques. We have applied this method to study the hERG K(+) channel and to determine the pharmacological profile of QT prolonging drugs.

  18. Inventing a co-axial atomic resolution patch clamp to study a single resonating protein complex and ultra-low power communication deep inside a living neuron cell.

    PubMed

    Ghosh, Subrata; Sahu, Satyajit; Agrawal, Lokesh; Shiga, Takashi; Bandyopadhyay, Anirban

    2016-12-01

    To read the signals of single molecules in vitro on a surface, or inside a living cell or organ, we introduce a coaxial atom tip (coat) and a coaxial atomic patch clamp (COAPAP). The metal-insulator-metal cavity of these probes extends to the atomic scale (0.1[Formula: see text]nm), it eliminates the cellular or environmental noise with a S/N ratio 10 5 . Five ac signals are simultaneously applied during a measurement by COAT and COAPAP to shield a true signal under environmental noise in five unique ways. The electromagnetic drive in the triaxial atomic tips is specifically designed to sense anharmonic vibrational and transmission signals for any system between 0.1[Formula: see text]nm and 50[Formula: see text]nm where the smallest nanopatch clamp cannot reach. COAT and COAPAP reliably pick up the atomic scale vibrations under the extreme noise of a living cell. Each protein's distinct electromagnetic, mechanical, electrical and ionic vibrational signature studied in vitro in a protected environment is found to match with the ones studied inside a live neuron. Thus, we could confirm that by using our probe blindly we could hold on to a single molecule or its complex in the invisible domain of a living cell. Our decade long investigations on perfecting the tools to measure bio-resonance of all forms and simultaneously in all frequency domains are summarized. It shows that the ratio of emission to absorption resonance frequencies of a biomaterial is around [Formula: see text], only a few in the entire em spectrum are active that regulates all other resonances, like mechanical, ionic, etc.

  19. M60 Tank Personnel Heater Comparison Test, Fort Carson, Colorado

    DTIC Science & Technology

    1983-05-01

    A study should be made considering the logisites and cost implications of developing a kit to upgrade the Model "C" heater to include self ...l f L Summary of Questionnaire Respose -for BLU (Model "A" with Engine Filter/Heat Tape) System Driver Loader Gunner TC’* " # % # % * % # 9 12. Heat

  20. 29 CFR 782.8 - Special classes of carriers.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ..., and ice) to railroad, docks, etc., for use of trains and steamships is not such transportation as is... cases cited in § 782.7(c), and see Mitchell v. Independent Ice Co., 294 F. 2d 186 (C.A. 5), certiorari..., driver's helpers, loaders, and mechanics employed by companies engaged in such activities are covered by...

  1. 29 CFR 782.8 - Special classes of carriers.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ..., and ice) to railroad, docks, etc., for use of trains and steamships is not such transportation as is... cases cited in § 782.7(c), and see Mitchell v. Independent Ice Co., 294 F. 2d 186 (C.A. 5), certiorari..., driver's helpers, loaders, and mechanics employed by companies engaged in such activities are covered by...

  2. 29 CFR 782.8 - Special classes of carriers.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., and ice) to railroad, docks, etc., for use of trains and steamships is not such transportation as is... cases cited in § 782.7(c), and see Mitchell v. Independent Ice Co., 294 F. 2d 186 (C.A. 5), certiorari..., driver's helpers, loaders, and mechanics employed by companies engaged in such activities are covered by...

  3. Chapter 5: Loosening compacted soils on mined lands

    Treesearch

    B. Strahm; R. Sweigard; J. Burger; D. Graves; C. Zipper; C. Barton; J. Skousen; P. Angel

    2017-01-01

    Successful surface coal mining businesses must move earth materials efficiently, so mining operations today depend on large and heavy equipment (Fig. 5-1). Track dozers and haul trucks used for mining can weigh more than 100 tons each. Wheel loaders and loaded haul trucks often exceed 200 tons. The mining industry has learned that successful reforestation of reclaimed...

  4. ARC-2008-ACD08-0217-001

    NASA Image and Video Library

    2008-09-24

    Aeronautics Technical Seminar series: Pilot Force Measurement with Inertia and Gravity Compensation by Rodger A. Mueller (offering an interesting behind-the-scenes look at some of the research that goes into creating high-fedelity, pilot-control loader simulation experiences for pilots and astronauts using the world renowned NASA Ames Vertical Motion Simulator) Series audio on file in Ames Library

  5. ARC-2008-ACD08-0217-002

    NASA Image and Video Library

    2008-09-24

    Aeroanutics Technical Seminar series: Pilot Force Measurement with Inertia and Gravity Compensation by Rodger A. Mueller (offering an interesting behind-the-scenes look at some of the research that goes into creating high-fedelity, pilot-control loader simulation experiences for pilots and astronauts using the world renowned NASA Ames Vertical Motion Simulator) Series audio on file in Ames Library

  6. ARC-2008-ACD08-0217-003

    NASA Image and Video Library

    2008-09-24

    Aeroanutics Technical Seminar series: Pilot Force Measurement with Inertia and Gravity Compensation by Rodger A. Mueller (offering an interesting behind-the-scenes look at some of the research that goes into creating high-fedelity, pilot-control loader simulation experiences for pilots and astronauts using the world renowned NASA Ames Vertical Motion Simulator) Series audio on file in Ames Library

  7. 76 FR 37795 - Notice of Availability of Government-Owned Inventions; Available for Licensing

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-28

    ... DEPARTMENT OF DEFENSE Department of the Navy Notice of Availability of Government-Owned Inventions....S. Patent No. 7,316,194: Rudders for High-Speed Ships//U.S. Patent No. 7,322,786: Mobile Loader for Transfer of Containers Between Delivery Vehicles and Marine Terminal Cranes//U.S. Patent No. 7,324,016...

  8. Department of Defense In-House RDT&E Activities

    DTIC Science & Technology

    1981-10-30

    DISEASES*.’......*......... 43 MISSILE LABORATORY ................................................. 44 MOBILITY EQUIPMENT R&D COMMAND...HELICOPTER INFLIGHT MONITORING SYS WITH INSTRUMENTED RANGE; ANECHOIC AND REVERBERATION CHAMBERS; MOBILE -’ ACCOUSTICS LAB; EYE MOVEMENT MEASURING...IN- CLUDES: LIGHTWEIGIHT OPTICAL TRACKING SYSTEM, 25-TON CRANE, 2 20-TON MOBILE CRANES, 5-TON WRECKER, 40,000 LB ACFT LOADER, THREE T-28B ACFT, 1 UH

  9. 77 FR 14495 - Certain New Pneumatic Off-the-Road Tires From the People's Republic of China: Final Results of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-12

    ... counterbalanced lift truck is a rigid framed, engine- powered machine with lift arms that has additional weight...; \\7\\ (2) construction vehicles and equipment, including earthmover articulated dump products, rigid frame haul trucks,\\8\\ front end loaders,\\9\\ dozers,\\10\\ lift trucks, straddle carriers,\\11\\ graders,\\12...

  10. Detachable clamps for minimal access surgery.

    PubMed

    Frank, T; Willetts, G J; Cuschieri, A

    1995-01-01

    A detachable clamp and applicator have been developed for use in minimal access surgical operations involving hollow visceral transection and anastomosis. The clamp has parallel jaws which ensure uniform distribution of the occlusive force. Following application on the bowel, the clamp is released from the applicator, thus freeing the access port. On completion of the anastomosis, the clamp is docked to the applicator, its jaws opened for release from the bowel and then closed prior to removal. The jaws of the clamp are kept closed by a pseudoelastic nickel-titanium (NiTi) alloy spring which imparts advantageous force characteristics when compared to stainless steel. The excellent holding and atraumatic characteristics of the detachable clamp have been confirmed by use in laparoscopic and thoracoscopic surgery on the gastrointestinal tract.

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

    Morris, John; Gilchrist, Phillip Charles

    Processes, systems, devices, and articles of manufacture are provided. Each may include adapting micro-inverters initially configured for frame-mounting to mounting on a frameless solar panel. This securement may include using an adaptive clamp or several adaptive clamps secured to a micro-inverter or its components, and using compressive forces applied directly to the solar panel to secure the adaptive clamp and the components to the solar panel. The clamps can also include compressive spacers and safeties for managing the compressive forces exerted on the solar panels. Friction zones may also be used for managing slipping between the clamp and the solarmore » panel during or after installation. Adjustments to the clamps may be carried out through various means and by changing the physical size of the clamps themselves.« less

  12. A novel high performance ESD power clamp circuit with a small area

    NASA Astrophysics Data System (ADS)

    Zhaonian, Yang; Hongxia, Liu; Li, Li; Qingqing, Zhuo

    2012-09-01

    A MOSFET-based electrostatic discharge (ESD) power clamp circuit with only a 10 ns RC time constant for a 0.18-μm process is proposed. A diode-connected NMOSFET is used to maintain a long delay time and save area. The special structure overcomes other shortcomings in this clamp circuit. Under fast power-up events, the gate voltage of the clamp MOSFET does not rise as quickly as under ESD events, the special structure can keep the clamp MOSFET thoroughly off. Under a falsely triggered event, the special structure can turn off the clamp MOSFET in a short time. The clamp circuit can also reject the power supply noise effectively. Simulation results show that the clamp circuit avoids fast false triggering events such as a 30 ns/1.8 V power-up, maintains a 1.2 μs delay time and a 2.14 μs turn-off time, and reduces to about 70% of the RC time constant. It is believed that the proposed clamp circuit can be widely used in high-speed integrated circuits.

  13. AstroML: Python-powered Machine Learning for Astronomy

    NASA Astrophysics Data System (ADS)

    Vander Plas, Jake; Connolly, A. J.; Ivezic, Z.

    2014-01-01

    As astronomical data sets grow in size and complexity, automated machine learning and data mining methods are becoming an increasingly fundamental component of research in the field. The astroML project (http://astroML.org) provides a common repository for practical examples of the data mining and machine learning tools used and developed by astronomical researchers, written in Python. The astroML module contains a host of general-purpose data analysis and machine learning routines, loaders for openly-available astronomical datasets, and fast implementations of specific computational methods often used in astronomy and astrophysics. The associated website features hundreds of examples of these routines being used for analysis of real astronomical datasets, while the associated textbook provides a curriculum resource for graduate-level courses focusing on practical statistics, machine learning, and data mining approaches within Astronomical research. This poster will highlight several of the more powerful and unique examples of analysis performed with astroML, all of which can be reproduced in their entirety on any computer with the proper packages installed.

  14. Drum ring removal/installation tool

    DOEpatents

    Andrade, William Andrew [Livermore, CA

    2006-11-14

    A handheld tool, or a pair of such tools, such as for use in removing/installing a bolt-type clamping ring on a container barrel/drum, where the clamping ring has a pair of clamping ends each with a throughbore. Each tool has an elongated handle and an elongated lever arm transversely connected to one end of the handle. The lever arm is capable of being inserted into the throughbore of a selected clamping end and leveraged with the handle to exert a first moment on the selected clamping end. Each tool also has a second lever arm, such as a socket with an open-ended slot, which is suspended alongside the first lever arm. The second lever arm is capable of engaging the selected clamping end and being leveraged with the handle to exert a second moment which is orthogonal to the first moment. In this manner, the first and second moments operate to hold the selected clamping end fixed relative to the tool so that the selected clamping end may be controlled with the handle. The pair of clamping ends may also be simultaneously and independently controlled with the use of two handles/tools so as to contort the geometry of the drum clamping ring and enable its removal/installation.

  15. Acute phase response in lactating dairy cows during hyperinsulinemic hypoglycaemic and hyperinsulinemic euglycaemic clamps and after intramammary LPS challenge.

    PubMed

    De Matteis, L; Bertoni, G; Lombardelli, R; Wellnitz, O; Van Dorland, H A; Vernay, M C M B; Bruckmaier, R M; Trevisi, E

    2017-06-01

    The link between energy availability, turnover of energy substrates and the onset of inflammation in dairy cows is complex and poorly investigated. To clarify this, plasma inflammatory variables were measured in mid-lactating dairy cows allocated to three groups: hyperinsulinemic hypoglycaemic clamp, induced by insulin infusion (HypoG, n = 5); hyperinsulinemic euglycaemic clamp, induced by insulin and glucose infusion (EuG; n = 6); control, receiving a saline solution infusion (NaCl; n = 6). At 48 h after the start of i.v. infusions, two udder quarters per cow were challenged with 200 μg of E. coli lipopolysaccharide (LPS). Individual blood samples were taken before clamps, before LPS challenge (i.e. 48 h after clamps) and 6.5 h after. At 48 h, positive acute phase proteins (posAPP) did not differ among groups, whereas albumin and cholesterol (index of lipoproteins), negative APP (negAPP), were lower (p < 0.05) in EuG compared to NaCl and HypoG. The concentration of IL-6 was greater in EuG (p < 0.05) but only vs. HypoG. At 6.5 h following LPS challenge, IL-6 increased in the NaCl and EuG clamps (p < 0.05), while TNF-α increased (p < 0.05) in the EuG only. Among the posAPP, haptoglobin markedly increased in EuG (p < 0.05), but not in NaCl (p = 0.76) and in HypoG; ceruloplasmin tended to decline during LPS challenge, the reduction was significant when all animals were considered (p < 0.05). Conversely, all the negAPP showed a marked reduction 6.5 h after LPS challenge in the three groups. In conclusion, EuG caused an inflammatory status after 48-h infusion (i.e. decrease of negAPP) and induced a quicker acute phase response (e.g. marked rise of TNF-α, IL-6) after the intramammary LPS challenge. These data suggest that the simultaneous high availability of glucose and insulin at the tissue-level makes dairy cows more susceptible to inflammatory events. In contrast, HypoG seems to attenuate the inflammatory response. Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.

  16. Welding fixture for joining bar-wound stator conductors

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

    De Souza, Urban J.; Rhoads, Frederick W.; Hanson, Justin

    A fixture assembly for welding a plurality of stator wire end pairs may include an anvil, a movable clamp configured to translate between an unclamped state and a clamped state, a first grounding electrode, and a second grounding electrode. The movable clamp may be configured to urge the plurality of stator wire ends against the anvil when in the clamped state. The moveable clamp includes a separator feature that generally extends toward the anvil. Each of the first grounding electrode and second grounding electrodes may be configured to translate between a clamped state and an unclamped state. When in themore » clamped state, each of the first and second grounding electrodes is configured to urge a pair of the plurality of stator wire end pairs against the separator feature.« less

  17. Physics-Based Design of Micro Air Vehicles

    DTIC Science & Technology

    2012-04-01

    7   Figure 5. Comparison of an insect wing and a manufactured wing for a flapping MAV. .............. 8...topologies for a flapping-wing compliant actuation mechanism. Hatched areas are clamped. Cases 1-3 have fixed supports; cases 4 and 5 have variable...world by flying insects , birds, and mammals. However, an inadequate understanding of the complex, nonlinear, and multidisciplinary physics that

  18. Real-Time linux dynamic clamp: a fast and flexible way to construct virtual ion channels in living cells.

    PubMed

    Dorval, A D; Christini, D J; White, J A

    2001-10-01

    We describe a system for real-time control of biological and other experiments. This device, based around the Real-Time Linux operating system, was tested specifically in the context of dynamic clamping, a demanding real-time task in which a computational system mimics the effects of nonlinear membrane conductances in living cells. The system is fast enough to represent dozens of nonlinear conductances in real time at clock rates well above 10 kHz. Conductances can be represented in deterministic form, or more accurately as discrete collections of stochastically gating ion channels. Tests were performed using a variety of complex models of nonlinear membrane mechanisms in excitable cells, including simulations of spatially extended excitable structures, and multiple interacting cells. Only in extreme cases does the computational load interfere with high-speed "hard" real-time processing (i.e., real-time processing that never falters). Freely available on the worldwide web, this experimental control system combines good performance. immense flexibility, low cost, and reasonable ease of use. It is easily adapted to any task involving real-time control, and excels in particular for applications requiring complex control algorithms that must operate at speeds over 1 kHz.

  19. Renal Vascular Clamp Placement: A Potential Cause of Incomplete Hilar Control during Partial Nephrectomy.

    PubMed

    Tryon, David; Myklak, Kristene; Alsyouf, Muhannad; Conceicao, Carol; Peplinski, Brandon; Arenas, Javier L; Faaborg, Daniel; Ruckle, Herbert C; Baldwin, D Duane

    2016-03-01

    Previous benchtop studies have shown that robotic bulldog clamps provide incomplete vascular control of a Penrose drain. We determined the efficacy of robotic and laparoscopic bulldog clamps to ensure hemostasis on the human renal artery. The effect of clamp position on vascular control was also examined. Fresh human cadaveric renal arteries were used to determine the leak point pressure of 7 bulldog clamps from a total of 3 manufacturers. Five trials were performed per clamp at 4 locations, including the fulcrum, proximal, middle and distal positions. Comparison was done using the Kruskal-Wallis test with p <0.05 considered significant. None of the bulldog clamps leaked at a pressure less than 215 mm Hg when applied at the proximal, middle or distal position. In general leak point pressure decreased as the artery was positioned more distal along the clamp. The exception was when the vessel was placed at the fulcrum position. At that position 80% to 100% of trials with the Klein laparoscopic, 100% with the Klein robotic (Klein Robotic, San Antonio, Texas) and 60% to 80% with the Scanlan robotic (Scanlan International, Saint Paul, Minnesota) clamp leaked at pressure below 215 mm Hg. Each vascular clamp adequately occluded flow at physiological pressure when placed at the proximal, middle or distal position. Furthermore, these results demonstrate that there is leakage at physiological pressure when the artery is placed at the fulcrum of certain clamp types. These results suggest that applying a bulldog clamp at the fulcrum could potentially lead to inadequate vessel occlusion and intraoperative bleeding. Copyright © 2016 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

  20. A comparison between the minimal model and the glucose clamp in the assessment of insulin sensitivity across the spectrum of glucose tolerance. Insulin Resistance Atherosclerosis Study.

    PubMed

    Saad, M F; Anderson, R L; Laws, A; Watanabe, R M; Kades, W W; Chen, Y D; Sands, R E; Pei, D; Savage, P J; Bergman, R N

    1994-09-01

    An insulin-modified frequently sampled intravenous glucose tolerance test (FSIGTT) with minimal model analysis was compared with the glucose clamp in 11 subjects with normal glucose tolerance (NGT), 20 with impaired glucose tolerance (IGT), and 24 with non-insulin-dependent diabetes mellitus (NIDDM). The insulin sensitivity index (SI) was calculated from FSIGTT using 22- and 12-sample protocols (SI(22) and SI(12), respectively). Insulin sensitivity from the clamp was expressed as SI(clamp) and SIP(clamp). Minimal model parameters were similar when calculated with SI(22) and SI(12). SI could not be distinguished from 0 in approximately 50% of diabetic patients with either protocol. SI(22) correlated significantly with SI(clamp) in the whole group (r = 0.62), and in the NGT (r = 0.53), IGT (r = 0.48), and NIDDM (r = 0.41) groups (P < 0.05 for each). SI(12) correlated significantly with SI(clamp) in the whole group (r = 0.55, P < 0.001) and in the NGT (r = 0.53, P = 0.046) and IGT (r = 0.58, P = 0.008) but not NIDDM (r = 0.30, P = 0.085) groups. When SI(22), SI(clamp), and SIP(clamp) were expressed in the same units, SI(22) was 66 +/- 5% (mean +/- SE) and 50 +/- 8% lower than SI(clamp) and SIP(clamp), respectively. Thus, minimal model analysis of the insulin-modified FSIGTT provides estimates of insulin sensitivity that correlate significantly with those from the glucose clamp. The correlation was weaker, however, in NIDDM. The insulin-modified FSIGTT can be used as a simple test for assessment of insulin sensitivity in population studies involving nondiabetic subjects. Additional studies are needed before using this test routinely in patients with NIDDM.

  1. Arterial waves in humans during peripheral vascular surgery.

    PubMed

    Khir, A W; Henein, M Y; Koh, T; Das, S K; Parker, K H; Gibson, D G

    2001-12-01

    The purpose of this study was to investigate the effect of aortic clamping on arterial waves during peripheral vascular surgery. We measured pressure and velocity simultaneously in the ascending aorta, in ten patients (70+/-5 years) with aortic-iliac disease intra-operatively. Pressure was measured using a catheter tip manometer, and velocity was measured using Doppler ultrasound. Data were collected before aortic clamping, during aortic clamping and after unclamping. Hydraulic work in the aortic root was calculated from the measured data, the reflected waves were determined by wave-intensity analysis and wave speed was determined by the PU-loop (pressure-velocity-loop) method; a new technique based on the 'water-hammer' equation. The wave speed is approx. 32% (P<0.05) higher during clamping than before clamping. Although the peak intensity of the reflected wave does not alter with clamping, it arrives 30 ms (P<0.05) earlier and its duration is 25% (P<0.05) longer than before clamping. During clamping, left ventricule (LV) hydraulic systolic work and the energy carried by the reflected wave increased by 27% (P<0.05) and 20% (P<0.05) respectively, compared with before clamping. The higher wave speed during clamping explains the earlier arrival of the reflected waves suggesting an increase in the afterload, since the LV has to overcome earlier reflected compression waves. The longer duration of the reflected wave during clamping is associated with an increase in the total energy carried by the wave, which causes an increase in hydraulic work. Increased hydraulic work during clamping may increase LV oxygen consumption, provoke myocardial ischaemia and hence contribute to the intra-operative impairment of LV function known in patients with peripheral vascular disease.

  2. [Microsurgical 2nd toe transfer for catastrophic hand reconstruction].

    PubMed

    Placer, A; Lozano, Ja

    2007-01-01

    The correct reconstruction of the catastrophic hand requires complex surgical techniques. The microsurgical transference of a toe is indicated when all other reconstructive options are shown to be useless for the reconstruction of the required clamp function. In this clinical note we set out the case of a 32 year old man, who came to our accident and emergency department after suffering a traffic accident. After exploration the diagnosis was that of catastrophic left hand, among other policontusions. Urgent surgery was carried out, saving the maximum possible viable structures. The immediate result of this surgery was a hand with 1st, 4th and 5th functional fingers. As the essential clamp function between the 1st and 4th or 5th fingers was not totally satisfactory, we decided to reconstruct the 3rd finger of his hand with his ipsilateral 2nd toe. All pertinent studies to determine vascularisation of the flap were carried out in planning the surgery, and the microsurgical transfer was then realized, which was successful. Today, after a suitable rehabilitation, the patient has recovered a satisfactory function of heavy and fine clamp in the operated hand. Toe to hand transfer is a good option for finger reconstruction and its function. Rehabilitation is the key to functional recovery.

  3. Obtaining spheroplasts of armored dinoflagellates and first single-channel recordings of their ion channels using patch-clamping.

    PubMed

    Pozdnyakov, Ilya; Matantseva, Olga; Negulyaev, Yuri; Skarlato, Sergei

    2014-09-05

    Ion channels are tightly involved in various aspects of cell physiology, including cell signaling, proliferation, motility, endo- and exo-cytosis. They may be involved in toxin production and release by marine dinoflagellates, as well as harmful algal bloom proliferation. So far, the patch-clamp technique, which is the most powerful method to study the activity of ion channels, has not been applied to dinoflagellate cells, due to their complex cellulose-containing cell coverings. In this paper, we describe a new approach to overcome this problem, based on the preparation of spheroplasts from armored bloom-forming dinoflagellate Prorocentrum minimum. We treated the cells of P. minimum with a cellulose synthesis inhibitor, 2,6-dichlorobenzonitrile (DCB), and found out that it could also induce ecdysis and arrest cell shape maintenance in these microalgae. Treatment with 100-250 µM DCB led to an acceptable 10% yield of P. minimum spheroplasts and was independent of the incubation time in the range of 1-5 days. We show that such spheroplasts are suitable for patch-clamping in the cell-attached mode and can form 1-10 GOhm patch contact with a glass micropipette, allowing recording of ion channel activity. The first single-channel recordings of dinoflagellate ion channels are presented.

  4. Obtaining Spheroplasts of Armored Dinoflagellates and First Single-Channel Recordings of Their Ion Channels Using Patch-Clamping

    PubMed Central

    Pozdnyakov, Ilya; Matantseva, Olga; Negulyaev, Yuri; Skarlato, Sergei

    2014-01-01

    Ion channels are tightly involved in various aspects of cell physiology, including cell signaling, proliferation, motility, endo- and exo-cytosis. They may be involved in toxin production and release by marine dinoflagellates, as well as harmful algal bloom proliferation. So far, the patch-clamp technique, which is the most powerful method to study the activity of ion channels, has not been applied to dinoflagellate cells, due to their complex cellulose-containing cell coverings. In this paper, we describe a new approach to overcome this problem, based on the preparation of spheroplasts from armored bloom-forming dinoflagellate Prorocentrum minimum. We treated the cells of P. minimum with a cellulose synthesis inhibitor, 2,6-dichlorobenzonitrile (DCB), and found out that it could also induce ecdysis and arrest cell shape maintenance in these microalgae. Treatment with 100–250 µM DCB led to an acceptable 10% yield of P. minimum spheroplasts and was independent of the incubation time in the range of 1–5 days. We show that such spheroplasts are suitable for patch-clamping in the cell-attached mode and can form 1–10 GOhm patch contact with a glass micropipette, allowing recording of ion channel activity. The first single-channel recordings of dinoflagellate ion channels are presented. PMID:25199048

  5. Clamp-mount device

    NASA Technical Reports Server (NTRS)

    Clark, K. H. (Inventor)

    1983-01-01

    A clamp-mount device is disclosed for mounting equipment to an associated I-beam and the like structural member of the type having oppositely extending flanges wherein the device comprises a base and a pair of oppositely facing clamping members carried diagonally on the base clamping flanges therebetween and having flange receiving openings facing one another. Lock means are carried diagonally by the base opposite the clamping members locking the flanges in the clamping members. A resilient hub is carried centrally of the base engaging and biasing a back side of the flanges maintaining tightly clamped and facilitating use on vertical as well as horizontal members. The base turns about the hub to receive the flanges within the clamping members. Equipment may be secured to the base by any suitable means such as bolts in openings. Slidable gate latches secure the hinged locks in an upright locking position. The resilient hub includes a recess opening formed in the base and a rubber-like pad carried in this opening being depressably and rotatably carried therein.

  6. Evaluation of Introduction of a Delayed Cord Clamping Protocol for Premature Neonates in a High-Volume Maternity Center.

    PubMed

    Liu, Lilly Y; Feinglass, Joe M; Khan, Janine Y; Gerber, Susan E; Grobman, William A; Yee, Lynn M

    2017-05-01

    To evaluate adherence to a delayed cord clamping protocol for preterm births in the first 2 years after its introduction, perform a quality improvement assessment, and determine neonatal outcomes associated with protocol implementation and adherence. This is a retrospective cohort study of women delivering singleton neonates at 23-32 weeks of gestation in the 2 years before (preprotocol) and 2 years after (postprotocol) introduction of a 30-second delayed cord clamping protocol at a large-volume academic center. This policy was communicated to obstetric and pediatric health care providers and nurses and reinforced with intermittent educational reviews. Barriers to receiving delayed cord clamping were assessed using χ tests and multivariable logistic regression. Neonatal outcomes then were compared between all neonates in the preprotocol period and all neonates in the postprotocol period and between all neonates in the preprotocol period and neonates receiving delayed cord clamping in the postprotocol period using multivariable linear and logistic regression analyses. Of the 427 eligible neonates, 187 were born postprotocol. Of these, 53.5% (n=100) neonates received delayed cord clamping according to the protocol. The rate of delayed cord clamping preprotocol was 0%. Protocol uptake and frequency of delayed cord clamping increased over the 2 years after its introduction. In the postprotocol period, cesarean delivery was the only factor independently associated with failing to receive delayed cord clamping (adjusted odds ratio [OR] 0.49, 95% confidence interval [CI] 0.25-0.96). In comparison with the preprotocol period, those who received delayed cord clamping in the postprotocol period had significantly higher birth hematocrit (β=2.46, P=.007) and fewer blood transfusions in the first week of life (adjusted OR 0.49, 95% CI 0.25-0.96). After introduction of an institutional delayed cord clamping protocol followed by continued health care provider education and quality feedback, the frequency of delayed cord clamping progressively increased. Compared with historical controls, performing delayed cord clamping in eligible preterm neonates was associated with improved neonatal hematologic indices, demonstrating the effectiveness of delayed cord clamping in a large-volume maternity unit.

  7. A clamped rectangular plate containing a crack

    NASA Technical Reports Server (NTRS)

    Tang, R.; Erdogan, F.

    1985-01-01

    The general problem of a rectangular plate clamped along two parallel sides and containing a crack parallel to the clamps is considered. The problem is formulated in terms of a system of singular integral equations and the asymptotic behavior of the stress state near the corners is investigated. Numerical examples are considered for a clamped plate without a crack and with a centrally located crack, and the stress intensity factors and the stresses along the clamps are calculated.

  8. Force-Measuring Clamp

    NASA Technical Reports Server (NTRS)

    Nunnelee, Mark (Inventor)

    2004-01-01

    A precision clamp that accurately measures force over a wide range of conditions is described. Using a full bridge or other strain gage configuration. the elastic deformation of the clamp is measured or detected by the strain gages. Thc strain gages transmit a signal that corresponds to the degree of stress upon the clamp. Thc strain gage signal is converted to a numeric display. Calibration is achieved by ero and span potentiometers which enable accurate measurements by the force-measuring clamp.

  9. Delayed clamping of the umbilical cord after delivery and implications for public cord blood banking.

    PubMed

    Allan, David S; Scrivens, Nicholas; Lawless, Tiffany; Mostert, Karen; Oppenheimer, Lawrence; Walker, Mark; Petraszko, Tanya; Elmoazzen, Heidi

    2016-03-01

    Public banking of umbilical cord blood units (CBUs) containing higher numbers of cells ensures timely engraftment after transplantation for increasing numbers of patients. Delayed clamping of the umbilical cord after birth may benefit some infants by preventing iron deficiency. Implications of delayed cord clamping for public cord blood banking remains unclear. CBUs collected by Canadian Blood Services at one collection site between November 1, 2014, and March 17, 2015, were analyzed. The delay in cord clamping after birth was timed and classified as "no delay," 20 to 60 seconds, more than 60 seconds, or more than 120 seconds. Of 367 collections, 100 reported no delay in clamping while clamping was delayed by 20 to 60 seconds (n = 69), more than 60 seconds (n = 98), or more than 120 seconds (n = 100) in the remaining cases. The mean volume and total nucleated cells (TNCs) in units with no delay in clamping were significantly greater than mean volumes for all categories of delayed clamping (Tukey's test, p < 0.05 for each comparison). The proportion of units with more than 1.5 × 10(9) TNCs was significantly reduced when clamping was delayed (p = 5.5 × 10(-8) ). The difference was most marked for cords that were clamped more than 120 seconds after delivery (6.2% compared with 39%). Delayed cord clamping greatly diminishes the volume and TNC count of units collected for a public cord blood bank. Creating an inventory of CBUs with high TNC content may take more time than expected. © 2015 AABB.

  10. 30 CFR 75.605 - Clamping of trailing cables to equipment.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Clamping of trailing cables to equipment. 75... MINE SAFETY AND HEALTH MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Trailing Cables § 75.605 Clamping of trailing cables to equipment. [Statutory Provisions] Trailing cables shall be clamped to...

  11. 76 FR 62356 - Certain New Pneumatic Off-the-Road Tires From the People's Republic of China: Preliminary Results...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-07

    ...-steer loaders are four-wheel drive vehicles with the left-side drive wheels independent of the right... exporter can demonstrate that it is sufficiently independent so as to be entitled to a separate rate.\\28... separate rate analysis is not necessary to determine whether it is independent from government control.\\29...

  12. Environmental Assessment: Anti-Terrorism/Force Protection McConnell Air Force Base, Kansas

    DTIC Science & Technology

    2003-09-01

    handled, stored, transported, disposed, or recycled in accordance with these regulations. The potential for hazardous waste generation from gate...Loader (rubber tire) Concrete Truck Concrete Finisher Crane Asphalt Spreader Roller Flat Bed Truck (18 wheel) Scraper Trenching Machine 1...plastics, and lumber. These materials would be placed in the appropriate construction materials landfill or recycled when possible. These wastes

  13. The Weak Link HP-41C hand-held calculator program

    Treesearch

    Ross A. Phillips; Penn A. Peters; Gary D. Falk

    1982-01-01

    The Weak Link hand-held calculator program (HP-41C) quickly analyzes a system for logging production and costs. The production equations model conventional chain saw, skidder, loader, and tandemaxle truck operations in eastern mountain areas. Production of each function of the logging system may be determined so that the system may be balanced for minimum cost. The...

  14. Comparison of two thinning systems. Part 1. Stand and site impacts

    Treesearch

    Bobby L. Lanford; Bryce J. Stokes

    1995-01-01

    During the winter of 1991, a side-by-side comparison was made between two popular thinning systems:a feller-buncher, grapple skidder, loader/slasher system and a harvester, forwarder system. A first commercial thinning was conducted in an 18-year-old loblolly pine stand Test areas were cruised prior to thinning and remeasured after operations were completed The target...

  15. Development and testing of a computer assisted remote-control system for the compact loader-trammer. Report of Investigations/1992

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

    Ruff, T.M.

    1992-01-01

    A prototype mucking machine designed to operate in narrow vein stopes was developed by Foster-Miller, Inc., Waltham, MA, under contract with the U.S. Bureau of Mines. The machine, called a compact loader/trammer, or minimucker, was designed to replace slusher muckers in narrow-vein underground mines. The minimucker is a six-wheel-drive, skid-steered, load-haul-dump machine that loads muck at the front with a novel slide-bucket system and ejects it out the rear so that the machine does not have to be turned around. To correct deficiencies of the tether remote control system, a computer-based, radio remote control was retrofitted to the minimucker. Initialmore » tests indicated a need to assist the operator in guiding the machine in narrow stopes and an automatic guidance system that used ultrasonic ranging sensors and a wall-following algorithm was installed. Additional tests in a simulated test stope showed that these changes improved the operation of the minimucker. The design and functions of the minimucker and its computer-based, remote control system are reviewed, and an ultrasonic, sensor-based guidance system is described.« less

  16. Development of the Los Alamos National Laboratory Cryogenic Pressure Loader

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

    Ebey, Peter S.; Dole, James M.; Hoffer, James K.

    2003-05-15

    Targets for inertial fusion research and ignition at OMEGA, the National Ignition Facility, LMJ, and future facilities rely on beta-radiation-driven layering of spherical cryogenic DT ice layers contained within plastic or metal shells. Plastic shells will be permeation filled at room temperature then cooled to cryogenic temperatures before removal of the overpressure. The cryogenic pressure loader (CPL) was recently developed at Los Alamos National Laboratory as a testbed for studying the filling and layering of plastic target shells with DT. A technical description of the CPL is provided. The CPL consists of a cryostat, which contains a high-pressure permeation cell,more » and has optical access for investigating beta layering. The cryostat is housed within a tritium glovebox that contains manifolds for supplying high-pressure DT. The CPL shares some design elements with the cryogenic target handling system at the OMEGA facility to allow testing of tritium issues related to that system. The CPL has the capability to fill plastic targets by permeation to pressures up to 100 MPa and to cool them to 15 K. The CPL will accommodate a range of targets and may be modified for future experiments.« less

  17. A practical tool for monitoring the performance of measuring systems in a laboratory network: report of an ACB Working Group.

    PubMed

    Ayling, Pete; Hill, Robert; Jassam, Nuthar; Kallner, Anders; Khatami, Zahra

    2017-11-01

    Background A logical consequence of the introduction of robotics and high-capacity analysers has seen a consolidation to larger units. This requires new structures and quality systems to ensure that laboratories deliver consistent and comparable results. Methods A spreadsheet program was designed to accommodate results from up to 12 different instruments/laboratories and present IQC data, i.e. Levey-Jennings and Youden plots and comprehensive numerical tables of the performance of each item. Input of data was made possible by a 'data loader' by which IQC data from the individual instruments could be transferred to the spreadsheet program on line. Results A set of real data from laboratories is used to populate the data loader and the networking software program. Examples are present from the analysis of variance components, the Levey-Jennings and Youden plots. Conclusions This report presents a software package that allows the simultaneous management and detailed monitoring of the performance of up to 12 different instruments/laboratories in a fully interactive mode. The system allows a quality manager of networked laboratories to have a continuous updated overview of the performance. This software package has been made available at the ACB website.

  18. Sliding Clamp of DNA Polymerase III as a Drug Target for TB Therapy: Comprehensive Conformational and Binding Analysis from Molecular Dynamic Simulations.

    PubMed

    Machaba, Kgothatso E; Cele, Favorite N; Mhlongo, Ndumiso N; Soliman, Mahmoud E S

    2016-12-01

    Tuberculosis, caused by Mycobacterium tuberculosis, is one of the most common causes of death in the world. Mycobacterium tuberculosis -sliding clamp is a protein essential for many important DNA transactions including replication and DNA repair proteins, thus, a potential drug target for tuberculosis. Further investigation is needed in understanding DNA polymerase sliding clamp structure, especially from a computational perspective. In this study, we employ a wide-range of comparative molecular dynamic analyses on two systems: Mycobacterium tuberculosis - sliding clamp enzyme in its apo and bound form. The results reported in this study shows apo conformation to be less stable, as compared to bound conformation with an average radius of gyration of 25.812 and 25.459 Å, respectively. This was further supported by root mean square fluctuation, where an apo enzyme showed a higher degree of flexibility. However, the presence of the ligand lowers radius of gyration and root mean square fluctuation and also leads to an existence of negative correlated motions. Principal component analysis further justifies the same findings, whereby the apo enzyme exhibits a higher fluctuation compared to the bound complex. In addition, a stable 3 10 helix located at the binding site appears to be unstable in the presence of the ligand. Hence, it is possible that the binding of the ligand may have caused a rearrangement of the structure, leading to a change in the unwinding of 3 10 helix. Findings reported in this study further enhance the understanding of Mycobacterium tuberculosis -DnaN and also give a lead to the development of potent tuberculosis drugs.

  19. Mechanical support of a ceramic gas turbine vane ring

    DOEpatents

    Shi, Jun; Green, Kevin E.; Mosher, Daniel A.; Holowczak, John E.; Reinhardt, Gregory E.

    2010-07-27

    An assembly for mounting a ceramic turbine vane ring onto a turbine support casing comprises a first metal clamping ring and a second metal clamping ring. The first metal clamping ring is configured to engage with a first side of a tab member of the ceramic turbine vane ring. The second metal clamping ring is configured to engage with a second side of the tab member such that the tab member is disposed between the first and second metal clamping rings.

  20. Active energy recovery clamping circuit to improve the performance of power converters

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

    Whitaker, Bret; Barkley, Adam

    2017-05-09

    A regenerative clamping circuit for a power converter using clamping diodes to transfer charge to a clamping capacitor and a regenerative converter to transfer charge out of the clamping capacitor back to the power supply input connection. The regenerative converter uses a switch connected to the midpoint of a series connected inductor and capacitor. The ends of the inductor and capacitor series are connected across the terminals of the power supply to be in parallel with the power supply.

  1. Payload Launch Lock Mechanism

    NASA Technical Reports Server (NTRS)

    Young, Ken (Inventor); Hindle, Timothy (Inventor)

    2014-01-01

    A payload launch lock mechanism includes a base, a preload clamp, a fastener, and a shape memory alloy (SMA) actuator. The preload clamp is configured to releasibly restrain a payload. The fastener extends, along an axis, through the preload clamp and into the base, and supplies a force to the preload clamp sufficient to restrain the payload. The SMA actuator is disposed between the base and the clamp. The SMA actuator is adapted to receive electrical current and is configured, upon receipt of the electrical current, to supply a force that causes the fastener to elongate without fracturing. The preload clamp, in response to the fastener elongation, either rotates or pivots to thereby release the payload.

  2. 21 CFR 876.5160 - Urological clamp for males.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Urological clamp for males. 876.5160 Section 876...) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5160 Urological clamp for males. (a) Identification. A urological clamp for males is a device used to close the urethra of a male to...

  3. Experimental Modal Analysis of Rectangular and Circular Beams

    ERIC Educational Resources Information Center

    Emory, Benjamin H.; Zhu, Wei Dong

    2006-01-01

    Analytical and experimental methods are used to determine the natural frequencies and mode shapes of Aluminum 6061-T651 beams with rectangular and circular cross-sections. A unique test stand is developed to provide the rectangular beam with different boundary conditions including clamped-free, clamped-clamped, clamped-pinned, and pinned-pinned.…

  4. Solutions for transients in arbitrarily branching cables: III. Voltage clamp problems.

    PubMed

    Major, G

    1993-07-01

    Branched cable voltage recording and voltage clamp analytical solutions derived in two previous papers are used to explore practical issues concerning voltage clamp. Single exponentials can be fitted reasonably well to the decay phase of clamped synaptic currents, although they contain many underlying components. The effective time constant depends on the fit interval. The smoothing effects on synaptic clamp currents of dendritic cables and series resistance are explored with a single cylinder + soma model, for inputs with different time courses. "Soma" and "cable" charging currents cannot be separated easily when the soma is much smaller than the dendrites. Subtractive soma capacitance compensation and series resistance compensation are discussed. In a hippocampal CA1 pyramidal neurone model, voltage control at most dendritic sites is extremely poor. Parameter dependencies are illustrated. The effects of series resistance compound those of dendritic cables and depend on the "effective capacitance" of the cell. Plausible combinations of parameters can cause order-of-magnitude distortions to clamp current waveform measures of simulated Schaeffer collateral inputs. These voltage clamp problems are unlikely to be solved by the use of switch clamp methods.

  5. Dynamic analysis of clamp band joint system subjected to axial vibration

    NASA Astrophysics Data System (ADS)

    Qin, Z. Y.; Yan, S. Z.; Chu, F. L.

    2010-10-01

    Clamp band joints are commonly used for connecting circular components together in industry. Some of the systems jointed by clamp band are subjected to dynamic load. However, very little research on the dynamic characteristics for this kind of joint can be found in the literature. In this paper, a dynamic model for clamp band joint system is developed. Contact and frictional slip between the components are accommodated in this model. Nonlinear finite element analysis is conducted to identify the model parameters. Then static experiments are carried out on a scaled model of the clamp band joint to validate the joint model. Finally, the model is adopted to study the dynamic characteristics of the clamp band joint system subjected to axial harmonic excitation and the effects of the wedge angle of the clamp band joint and the preload on the response. The model proposed in this paper can represent the nonlinearity of the clamp band joint and be used conveniently to investigate the effects of the structural and loading parameters on the dynamic characteristics of this type of joint system.

  6. Application of active electrode compensation to perform continuous voltage-clamp recordings with sharp microelectrodes.

    PubMed

    Gómez-González, J F; Destexhe, A; Bal, T

    2014-10-01

    Electrophysiological recordings of single neurons in brain tissues are very common in neuroscience. Glass microelectrodes filled with an electrolyte are used to impale the cell membrane in order to record the membrane potential or to inject current. Their high resistance induces a high voltage drop when passing current and it is essential to correct the voltage measurements. In particular, for voltage clamping, the traditional alternatives are two-electrode voltage-clamp technique or discontinuous single electrode voltage-clamp (dSEVC). Nevertheless, it is generally difficult to impale two electrodes in a same neuron and the switching frequency is limited to low frequencies in the case of dSEVC. We present a novel fully computer-implemented alternative to perform continuous voltage-clamp recordings with a single sharp-electrode. To reach such voltage-clamp recordings, we combine an active electrode compensation algorithm (AEC) with a digital controller (AECVC). We applied two types of control-systems: a linear controller (proportional plus integrative controller) and a model-based controller (optimal control). We compared the performance of the two methods to dSEVC using a dynamic model cell and experiments in brain slices. The AECVC method provides an entirely digital method to perform continuous recording and smooth switching between voltage-clamp, current clamp or dynamic-clamp configurations without introducing artifacts.

  7. 7. COMPLETE X15 VEHICLE TEST STAND AFTER AN ENGINE FIRE ...

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

    7. COMPLETE X-15 VEHICLE TEST STAND AFTER AN ENGINE FIRE OR EXPLOSION. Wreckage of engine is still fixed in its clamp; X-15 vehicle lies on the ground detached from engine. - Edwards Air Force Base, X-15 Engine Test Complex, Rocket Engine & Complete X-15 Vehicle Test Stands, Rogers Dry Lake, east of runway between North Base & South Base, Boron, Kern County, CA

  8. Protection of the vehicle cab environment against bacteria, fungi and endotoxins in composting facilities.

    PubMed

    Schlosser, O; Huyard, A; Rybacki, D; Do Quang, Z

    2012-06-01

    Microbial quality of air inside vehicle cabs is a major occupational health risk management issue in composting facilities. Large differences and discrepancies in protection factors between vehicles and between biological agents have been reported. This study aimed at estimating the mean protection efficiency of the vehicle cab environment against bioaerosols with higher precision. In-cab measurement results were also analysed to ascertain whether or not these protection systems reduce workers' exposure to tolerable levels. Five front-end loaders, one mobile mixer and two agricultural tractors pulling windrow turners were investigated. Four vehicles were fitted with a pressurisation and high efficiency particulate air (HEPA) filtration system. The four others were only equipped with pleated paper filter without pressurisation. Bacteria, fungi and endotoxins were measured in 72 pairs of air samples, simultaneously collected inside the cab and on the outside of the cab with a CIP 10-M sampler. A front-end loader, purchased a few weeks previously, fitted with a pressurisation and high efficiency particulate air (HEPA) filtration system, and with a clean cab, exhibited a mean protection efficiency of between 99.47% CI 95% [98.58-99.97%] and 99.91% [99.78-99.98%] depending on the biological agent. It is likely that the lower protection efficiency demonstrated in other vehicles was caused by penetration through the only moderately efficient filters, by the absence of pressurisation, by leakage in the filter-sealing system, and by re-suspension of particles which accumulated in dirty cabs. Mean protection efficiency in regards to bacteria and endotoxins ranged between 92.64% [81.87-97.89%] and 98.61% [97.41-99.38%], and between 92.68% [88.11-96.08%] and 98.43% [97.44-99.22%], respectively. The mean protection efficiency was the lowest when confronted with fungal spores, from 59.76% [4.19-90.75%] to 94.71% [91.07-97.37%]. The probability that in-cab exposure to fungi exceeded the benchmark value for short-term respiratory effects suggests that front-end loaders and mobile mixers in composting facilities should be fitted with a pressurisation and HEPA filtration system, regardless of whether or not the facility is indoors or outdoors. Regarding the tractors, exposure inside the cabs was not significantly reduced. However, in this study, there was a less than 0.01% risk of exceeding the bench mark value associated with fungi related short-term respiratory effects during an 1-h per day windrow turning operation. Pressurisation and a HEPA filtration system can provide safe working conditions inside loaders and mobile mixer with regard to airborne bacteria, fungi and endotoxins in composting facilities. However, regular thorough cleaning of the vehicle cab, as well as overalls and shoes cleaning, and mitigation of leakage in the filter-sealing system are necessary to achieve high levels of protection efficiency. Copyright © 2012 Elsevier Ltd. All rights reserved.

  9. Localized, stepwise template growth of functional nanowires from an amino acid-supported framework in a microfluidic chip.

    PubMed

    Puigmartí-Luis, Josep; Rubio-Martínez, Marta; Imaz, Inhar; Cvetković, Benjamin Z; Abad, Llibertat; Pérez Del Pino, Angel; Maspoch, Daniel; Amabilino, David B

    2014-01-28

    A spatially controlled synthesis of nanowire bundles of the functional crystalline coordination polymer (CP) Ag(I)TCNQ (tetracyanoquinodimethane) from previously fabricated and trapped monovalent silver CP (Ag(I)Cys (cysteine)) using a room-temperature microfluidic-assisted templated growth method is demonstrated. The incorporation of microengineered pneumatic clamps in a two-layer polydimethylsiloxane-based (PDMS) microfluidic platform was used. Apart from guiding the formation of the Ag(I)Cys coordination polymer, this microfluidic approach enables a local trapping of the in situ synthesized structures with a simple pneumatic clamp actuation. This method not only enables continuous and multiple chemical events to be conducted upon the trapped structures, but the excellent fluid handling ensures a precise chemical activation of the amino acid-supported framework in a position controlled by interface and clamp location that leads to a site-specific growth of Ag(I)TCNQ nanowire bundles. The synthesis is conducted stepwise starting with Ag(I)Cys CPs, going through silver metal, and back to a functional CP (Ag(I)TCNQ); that is, a novel microfluidic controlled ligand exchange (CP → NP → CP) is presented. Additionally, the pneumatic clamps can be employed further to integrate the conductive Ag(I)TCNQ nanowire bundles onto electrode arrays located on a surface, hence facilitating the construction of the final functional interfaced systems from solution specifically with no need for postassembly manipulation. This localized self-supported growth of functional matter from an amino acid-based CP shows how sequential localized chemistry in a fluid cell can be used to integrate molecular systems onto device platforms using a chip incorporating microengineered pneumatic tools. The control of clamp pressure and in parallel the variation of relative flow rates of source solutions permit deposition of materials at different locations on a chip that could be useful for device array preparation. The in situ reaction and washing procedures make this approach a powerful one for the fabrication of multicomponent complex nanomaterials using a soft bottom-up approach.

  10. Optimal Surgical Therapy in a Porcine (Sus scrofa) Model of Extra-Thoracic Penetrating Trauma Resulting in Hemorrhagic Shock: ED Thoracotomy vs. Immediate Trans-Abdominal Vascular Control. A Porcine Model for Evaluating the Management of Non-Compressible Torso Hemorrhage

    DTIC Science & Technology

    2010-11-08

    celiac aortic clamping (n=6), direct vascular control (n=6), and endovascular aortic occlusion n=6). This study presents a large animal model of class...including thoracic aortic clamping, supra- celiac aortic clamping, direct vascular control, and proximal endovascular balloon occlusion. Following vascular...subsequently underwent non-compressible hemorrhage with thoracic aortic clamping (n=6), supra- celiac aortic clamping (n=6), direct vascular control (n=6

  11. Device for remote operation of electrical disconnect

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

    Coleman, Jody Rustyn; Bobbitt, III, John Thomas

    Provided is a device for remote operation of an electrical disconnect. The device can include a handle clamp configured to be secured to an extending member of the electrical disconnect. The device can further include a case clamp configured to be secured to a rigid portion of the electrical disconnect. The device can further include a cable having an exterior sheath coaxially surrounding an inner cable. The inner cable can be coaxially slidable with respect to the exterior sheath. The inner cable can extend through an opening of the case clamp and be secured to the handle clamp. The devicemore » can further include an actuator configured to coaxially slide the inner cable such that the handle clamp is actuated towards the case clamp.« less

  12. 24. BUILDING NO. 452, ORDNANCE FACILITY (BAG CHARGE FILLING PLANT), ...

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

    24. BUILDING NO. 452, ORDNANCE FACILITY (BAG CHARGE FILLING PLANT), INTERIOR VIEW LOOKING WEST AT NORTH END OF CENTRAL CORRIDOR (ROOM 3). STAIRWAY WORKBENCH WITH COMPRESSED-AIR POWERED CARTRIDGE LOADER. ARMORED PASS-THROUGH OF TRANSFER BOX FOR PASSING EXPLOSIVES MATERIALS THROUGH TO NEXT ROOM TO THE NORTH. - Picatinny Arsenal, 400 Area, Gun Bag Loading District, State Route 15 near I-80, Dover, Morris County, NJ

  13. Conveyor-Loader Plastic Toy Parts (toys and games) 9-13.01--Technical Report on Standardization of the General Aptitude Test Battery.

    ERIC Educational Resources Information Center

    Manpower Administration (DOL), Washington, DC. U.S. Training and Employment Service.

    The United States Training and Employment Service General Aptitude Test Battery (GATB), first published in 1947, has been included in a continuing program of research to validate the tests against success in many different occupations. The GATB consists of 12 tests which measure nine aptitudes: General Learning Ability; Verbal Aptitude; Numerical…

  14. 29 CFR 782.5 - Loaders.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... parte Nos. MC-2 and MC-3, 28 M.C.C. 125, 133, 134, 139), is an employee of a carrier subject to section... U.S. 649; Pyramid Motor Freight Corp. v. Ispass, 330 U.S. 695; Walling v. Gordon's Transport (W.D...; Walling v. Huber & Huber Motor Express, 67 F. Supp. 855; Ex parte Nos. MC-2 and MC-3, 28 M.C.C. 125, 133...

  15. 29 CFR 782.5 - Loaders.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... parte Nos. MC-2 and MC-3, 28 M.C.C. 125, 133, 134, 139), is an employee of a carrier subject to section... U.S. 649; Pyramid Motor Freight Corp. v. Ispass, 330 U.S. 695; Walling v. Gordon's Transport (W.D...; Walling v. Huber & Huber Motor Express, 67 F. Supp. 855; Ex parte Nos. MC-2 and MC-3, 28 M.C.C. 125, 133...

  16. 29 CFR 782.5 - Loaders.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... parte Nos. MC-2 and MC-3, 28 M.C.C. 125, 133, 134, 139), is an employee of a carrier subject to section... U.S. 649; Pyramid Motor Freight Corp. v. Ispass, 330 U.S. 695; Walling v. Gordon's Transport (W.D...; Walling v. Huber & Huber Motor Express, 67 F. Supp. 855; Ex parte Nos. MC-2 and MC-3, 28 M.C.C. 125, 133...

  17. FPGA Boot Loader and Scrubber

    NASA Technical Reports Server (NTRS)

    Wade, Randall S.; Jones, Bailey

    2009-01-01

    A computer program loads configuration code into a Xilinx field-programmable gate array (FPGA), reads back and verifies that code, reloads the code if an error is detected, and monitors the performance of the FPGA for errors in the presence of radiation. The program consists mainly of a set of VHDL files (wherein "VHDL" signifies "VHSIC Hardware Description Language" and "VHSIC" signifies "very-high-speed integrated circuit").

  18. A structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation.

    PubMed

    Dai, Peng; Williams, Jonathan K; Zhang, Chi; Welborn, Matthew; Shepherd, James J; Zhu, Tianyu; Van Voorhis, Troy; Hong, Mei; Pentelute, Bradley L

    2017-08-11

    Natural enzymes use local environments to tune the reactivity of amino acid side chains. In searching for small peptides with similar properties, we discovered a four-residue π-clamp motif (Phe-Cys-Pro-Phe) for regio- and chemoselective arylation of cysteine in ribosomally produced proteins. Here we report mutational, computational, and structural findings directed toward elucidating the molecular factors that drive π-clamp-mediated arylation. We show the significance of a trans conformation prolyl amide bond for the π-clamp reactivity. The π-clamp cysteine arylation reaction enthalpy of activation (ΔH ‡ ) is significantly lower than a non-π-clamp cysteine. Solid-state NMR chemical shifts indicate the prolyl amide bond in the π-clamp motif adopts a 1:1 ratio of the cis and trans conformation, while in the reaction product Pro3 was exclusively in trans. In two structural models of the perfluoroarylated product, distinct interactions at 4.7 Å between Phe1 side chain and perfluoroaryl electrophile moiety are observed. Further, solution 19 F NMR and isothermal titration calorimetry measurements suggest interactions between hydrophobic side chains in a π-clamp mutant and the perfluoroaryl probe. These studies led us to design a π-clamp mutant with an 85-fold rate enhancement. These findings will guide us toward the discovery of small reactive peptides to facilitate abiotic chemistry in water.

  19. Combination Space Station Handrail Clamp and Pointing Device

    NASA Technical Reports Server (NTRS)

    Hughes, Stephen J. (Inventor)

    1999-01-01

    A device for attaching an experiment carrier to a space station handrail is provided. The device has two major components, a clamping mechanism for attachment to a space station handrail, and a pointing carrier on which an experiment package can be mounted and oriented. The handrail clamp uses an overcenter mechanism and the carrier mechanism uses an adjustable preload ball and socket for carrier positioning. The handrail clamp uses a stack of disk springs to provide a spring loaded button. This configuration provides consistent clamping force over a range of possible handrail thicknesses. Three load points are incorporated in the clamping mechanism thereby spreading the clamping load onto three separate points on the handrail. A four bar linkage is used to provide for a single actuation lever for all three load points. For additional safety, a secondary lock consisting of a capture plate and push lock keeps the clamp attached to the handrail in the event of main clamp failure. For the carrier positioning mechanism, a ball in a spring loaded socket uses friction to provide locking torque; however. the ball and socket are torque limited so that the ball ran slip under kick loads (125 pounds or greater). A lead screw attached to disk spring stacks is used to provide an adjustable spring force on the socket. A locking knob is attached to the lead screw to allow for hand manipulation of the lead screw.

  20. Structural and mechanistic insights into Mcm2-7 double-hexamer assembly and function

    DOE PAGES

    Sun, Jingchuan; Li, Huilin; Fernandez-Cid, Alejandra; ...

    2014-10-15

    Eukaryotic cells license each DNA replication origin during G1 phase by assembling a prereplication complex that contains a Mcm2–7 (minichromosome maintenance proteins 2–7) double hexamer. During S phase, each Mcm2–7 hexamer forms the core of a replicative DNA helicase. However, the mechanisms of origin licensing and helicase activation are poorly understood. The helicase loaders ORC–Cdc6 function to recruit a single Cdt1–Mcm2–7 heptamer to replication origins prior to Cdt1 release and ORC–Cdc6–Mcm2–7 complex formation, but how the second Mcm2–7 hexamer is recruited to promote double-hexamer formation is not well understood. Here, structural evidence for intermediates consisting of an ORC–Cdc6–Mcm2–7 complex andmore » an ORC–Cdc6–Mcm2–7–Mcm2–7 complex are reported, which together provide new insights into DNA licensing. Detailed structural analysis of the loaded Mcm2–7 double-hexamer complex demonstrates that the two hexamers are interlocked and misaligned along the DNA axis and lack ATP hydrolysis activity that is essential for DNA helicase activity. Moreover, we show that the head-to-head juxtaposition of the Mcm2–7 double hexamer generates a new protein interaction surface that creates a multisubunit-binding site for an S-phase protein kinase that is known to activate DNA replication. The data suggest how the double hexamer is assembled and how helicase activity is regulated during DNA licensing, with implications for cell cycle control of DNA replication and genome stability.« less

  1. Cardiovascular transition at birth: a physiological sequence.

    PubMed

    Hooper, Stuart B; Te Pas, Arjan B; Lang, Justin; van Vonderen, Jeroen J; Roehr, Charles Christoph; Kluckow, Martin; Gill, Andrew W; Wallace, Euan M; Polglase, Graeme R

    2015-05-01

    The transition to newborn life at birth involves major cardiovascular changes that are triggered by lung aeration. These include a large increase in pulmonary blood flow (PBF), which is required for pulmonary gas exchange and to replace umbilical venous return as the source of preload for the left heart. Clamping the umbilical cord before PBF increases reduces venous return and preload for the left heart and thereby reduces cardiac output. Thus, if ventilation onset is delayed following cord clamping, the infant is at risk of superimposing an ischemic insult, due to low cardiac output, on top of an asphyxic insult. Much debate has centered on the timing of cord clamping at birth, focusing mainly on the potential for a time-dependent placental to infant blood transfusion. This has prompted recommendations for delayed cord clamping for a set time after birth in infants not requiring resuscitation. However, recent evidence indicates that ventilation onset before cord clamping mitigates the adverse cardiovascular consequences caused by immediate cord clamping. This indicates that the timing of cord clamping should be based on the infant's physiology rather than an arbitrary period of time and that delayed cord clamping may be of greatest benefit to apneic infants.

  2. Planar patch clamp for neuronal networks--considerations and future perspectives.

    PubMed

    Bosca, Alessandro; Martina, Marzia; Py, Christophe

    2014-01-01

    The patch-clamp technique is generally accepted as the gold standard for studying ion channel activity allowing investigators to either "clamp" membrane voltage and directly measure transmembrane currents through ion channels, or to passively monitor spontaneously occurring intracellular voltage oscillations. However, this resulting high information content comes at a price. The technique is labor-intensive and requires highly trained personnel and expensive equipment. This seriously limits its application as an interrogation tool for drug development. Patch-clamp chips have been developed in the last decade to overcome the tedious manipulations associated with the use of glass pipettes in conventional patch-clamp experiments. In this chapter, we describe some of the main materials and fabrication protocols that have been developed to date for the production of patch-clamp chips. We also present the concept of a patch-clamp chip array providing high resolution patch-clamp recordings from individual cells at multiple sites in a network of communicating neurons. On this chip, the neurons are aligned with the aperture-probes using chemical patterning. In the discussion we review the potential use of this technology for pharmaceutical assays, neuronal physiology and synaptic plasticity studies.

  3. Design and experimental research of a novel inchworm type piezo-driven rotary actuator with the changeable clamping radius.

    PubMed

    Zhao, Hongwei; Fu, Lu; Ren, Luquan; Huang, Hu; Fan, Zunqiang; Li, Jianping; Qu, Han

    2013-01-01

    In this paper, a novel piezo-driven rotary actuator with the changeable clamping radius is developed based on the inchworm principle. This actuator mainly utilizes three piezoelectric actuators, a flexible gripper, a clamping block, and a rotor to achieve large stroke rotation with high resolution. The design process of the flexible gripper consisting of the driving unit and the clamping unit is described. Lever-type mechanisms were used to amplify the micro clamping displacements. The amplifying factor and parasitic displacement of the lever-type mechanism in the clamping unit was analyzed theoretically and experimentally. In order to investigate the rotation characteristics of the actuator, a series of experiments was carried out. Experimental results indicate that the actuator can rotate at a speed of 77,488 μrad/s with a driving frequency of 167 Hz. The rotation resolution and maximum load torque of the actuator are 0.25 μrad and 37 N mm, respectively. The gripper is movable along the z direction based on an elevating platform, and the clamping radius can change from 10.6 mm to 25 mm. Experimental results confirm that the actuator can achieve different rotation speeds by changing the clamping radius.

  4. Clinical aspects of incorporating cord clamping into stabilisation of preterm infants.

    PubMed

    Knol, Ronny; Brouwer, Emma; Vernooij, Alex S N; Klumper, Frans J C M; DeKoninck, Philip; Hooper, Stuart B; Te Pas, Arjan B

    2018-04-21

    Fetal to neonatal transition is characterised by major pulmonary and haemodynamic changes occurring in a short period of time. In the international neonatal resuscitation guidelines, comprehensive recommendations are available on supporting pulmonary transition and delaying clamping of the cord in preterm infants. Recent experimental studies demonstrated that the pulmonary and haemodynamic transition are intimately linked, could influence each other and that the timing of umbilical cord clamping should be incorporated into the respiratory stabilisation. We reviewed the current knowledge on how to incorporate cord clamping into stabilisation of preterm infants and the physiological-based cord clamping (PBCC) approach, with the infant's transitional status as key determinant of timing of cord clamping. This approach could result in optimal timing of cord clamping and has the potential to reduce major morbidities and mortality in preterm infants. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  5. Sound absorption by clamped poroelastic plates.

    PubMed

    Aygun, H; Attenborough, K

    2008-09-01

    Measurements and predictions have been made of the absorption coefficient and the surface acoustic impedance of poroelastic plates clamped in a large impedance tube and separated from the rigid termination by an air gap. The measured and predicted absorption coefficient and surface impedance spectra exhibit low frequency peaks. The peak frequencies observed in the absorption coefficient are close to those predicted and measured in the deflection spectra of the clamped poroelastic plates. The influences of the rigidity of the clamping conditions and the width of the air gap have been investigated. Both influences are found to be important. Increasing the rigidity of clamping reduces the low frequency absorption peaks compared with those measured for simply supported plates or plates in an intermediate clamping condition. Results for a closed cell foam plate and for two open cell foam plates made from recycled materials are presented. For identical clamping conditions and width of air gap, the results for the different materials differ as a consequence mainly of their different elasticity, thickness, and cell structure.

  6. Double arch mirror study. Part 1: Preliminary engineering report

    NASA Technical Reports Server (NTRS)

    Vukobratovich, D.; Hillman, D.

    1983-01-01

    In the proposed design, the NASA AMES 20-in double arch mirror is supported by three clamp and flexure assemblies. The mirror clamp consists of a T-shaped Invar-36 member that goes into a similarly shaped socket in the back of the mirror. The mirror socket is made oversize and contacts the clamp only along the conical surface. The clamp is preloaded by a spring washer and pulls the mirror into contact with the flexure. The clamp is then inserted into the mirror socket through a cutout, is rotated 90 deg, and is then pinned in place. Loading conditions considered in socket design are discussed as well as stress in the socket and clamp. Flexure geometry and stress are examined as well as the effects of flexure error and of mirror cell error.

  7. Fabrication and characterization of a piezoelectric energy harvester with clamped-clamped beams

    NASA Astrophysics Data System (ADS)

    Cui, Yan; Yu, Menglin; Gao, Shiqiao; Kong, Xiangxin; Gu, Wang; Zhang, Ran; Liu, Bowen

    2018-05-01

    This work presents a piezoelectric energy harvester with clamped-clamped beams, and it is fabricated with MEMS process. When excited by sinusoidal vibration, the energy harvester has a sharp jumping down phenomenon and the measured frequency responses of the clamped-clamped beams structure show a larger bandwidth which is about 56Hz, more efficient than that with cantilever beams. When the exciting acceleration ac is 12m/s2, the energy harvester achieves to a maximum open-circuit voltage of 94mV on one beam. The load voltage is proportional to the load resistance, and it increased with the increase of load resistance. Connected four beams in series, the output power reaches the maximum value of 730 nW and the optimal load is 15KΩ to one beam.

  8. Rigid clamp

    DOEpatents

    Benavides, G.L.; Burt, J.D.

    1994-07-12

    The invention relates to a clamp mechanism that can be used to attach or temporarily support objects inside of tubular goods. The clamp mechanism can also be modified so that it grips objects. The clamp has a self-centering feature to accommodate out-of-roundness or other internal defections in tubular objects such as pipe. A plurality of clamping shoes are expanded by a linkage which is preferably powered by a motor to contact the inside of a pipe. The motion can be reversed and jaw elements can be connected to the linkage so as to bring the jaws together to grab an object. 12 figs.

  9. Rigid clamp

    DOEpatents

    Benavides, Gilbert L.; Burt, Jack D.

    1994-01-01

    The invention relates to a clamp mechanism that can be used to attach or temporarily support objects inside of tubular goods. The clamp mechanism can also be modified so that it grips objects. The clamp has a self-centering feature to accommodate out-of-roundness or other internal defections in tubular objects such as pipe. A plurality of clamping shoes are expanded by a linkage which is preferably powered by a motor to contact the inside of a pipe. The motion can be reversed and jaw elements can be connected to the linkage so as to bring the jaws together to grab an object.

  10. Advanced motor driven clamped borehole seismic receiver

    DOEpatents

    Engler, Bruce P.; Sleefe, Gerard E.; Striker, Richard P.

    1993-01-01

    A borehole seismic tool including a borehole clamp which only moves perpendicular to the borehole. The clamp is driven by an electric motor, via a right angle drive. When used as a seismic receiver, the tool has a three part housing, two of which are hermetically sealed. Accelerometers or geophones are mounted in one hermetically sealed part, the electric meter in the other hermetically sealed part, and the clamp and right angle drive in the third part. Preferably the tool includes cable connectors at both ends. Optionally a shear plate can be added to the clamp to extend the range of the tool.

  11. Radial wedge flange clamp

    DOEpatents

    Smith, Karl H.

    2002-01-01

    A radial wedge flange clamp comprising a pair of flanges each comprising a plurality of peripheral flat wedge facets having flat wedge surfaces and opposed and mating flat surfaces attached to or otherwise engaged with two elements to be joined and including a series of generally U-shaped wedge clamps each having flat wedge interior surfaces and engaging one pair of said peripheral flat wedge facets. Each of said generally U-shaped wedge clamps has in its opposing extremities apertures for the tangential insertion of bolts to apply uniform radial force to said wedge clamps when assembled about said wedge segments.

  12. Novel screening techniques for ion channel targeting drugs

    PubMed Central

    Obergrussberger, Alison; Stölzle-Feix, Sonja; Becker, Nadine; Brüggemann, Andrea; Fertig, Niels; Möller, Clemens

    2015-01-01

    Ion channels are integral membrane proteins that regulate the flux of ions across the cell membrane. They are involved in nearly all physiological processes, and malfunction of ion channels has been linked to many diseases. Until recently, high-throughput screening of ion channels was limited to indirect, e.g. fluorescence-based, readout technologies. In the past years, direct label-free biophysical readout technologies by means of electrophysiology have been developed. Planar patch-clamp electrophysiology provides a direct functional label-free readout of ion channel function in medium to high throughput. Further electrophysiology features, including temperature control and higher-throughput instruments, are continually being developed. Electrophysiological screening in a 384-well format has recently become possible. Advances in chip and microfluidic design, as well as in cell preparation and handling, have allowed challenging cell types to be studied by automated patch clamp. Assays measuring action potentials in stem cell-derived cardiomyocytes, relevant for cardiac safety screening, and neuronal cells, as well as a large number of different ion channels, including fast ligand-gated ion channels, have successfully been established by automated patch clamp. Impedance and multi-electrode array measurements are particularly suitable for studying cardiomyocytes and neuronal cells within their physiological network, and to address more complex physiological questions. This article discusses recent advances in electrophysiological technologies available for screening ion channel function and regulation. PMID:26556400

  13. Novel screening techniques for ion channel targeting drugs.

    PubMed

    Obergrussberger, Alison; Stölzle-Feix, Sonja; Becker, Nadine; Brüggemann, Andrea; Fertig, Niels; Möller, Clemens

    2015-01-01

    Ion channels are integral membrane proteins that regulate the flux of ions across the cell membrane. They are involved in nearly all physiological processes, and malfunction of ion channels has been linked to many diseases. Until recently, high-throughput screening of ion channels was limited to indirect, e.g. fluorescence-based, readout technologies. In the past years, direct label-free biophysical readout technologies by means of electrophysiology have been developed. Planar patch-clamp electrophysiology provides a direct functional label-free readout of ion channel function in medium to high throughput. Further electrophysiology features, including temperature control and higher-throughput instruments, are continually being developed. Electrophysiological screening in a 384-well format has recently become possible. Advances in chip and microfluidic design, as well as in cell preparation and handling, have allowed challenging cell types to be studied by automated patch clamp. Assays measuring action potentials in stem cell-derived cardiomyocytes, relevant for cardiac safety screening, and neuronal cells, as well as a large number of different ion channels, including fast ligand-gated ion channels, have successfully been established by automated patch clamp. Impedance and multi-electrode array measurements are particularly suitable for studying cardiomyocytes and neuronal cells within their physiological network, and to address more complex physiological questions. This article discusses recent advances in electrophysiological technologies available for screening ion channel function and regulation.

  14. The DnaA N-terminal domain interacts with Hda to facilitate replicase clamp-mediated inactivation of DnaA.

    PubMed

    Su'etsugu, Masayuki; Harada, Yuji; Keyamura, Kenji; Matsunaga, Chika; Kasho, Kazutoshi; Abe, Yoshito; Ueda, Tadashi; Katayama, Tsutomu

    2013-12-01

    DnaA activity for replication initiation of the Escherichia coli chromosome is negatively regulated by feedback from the DNA-loaded form of the replicase clamp. In this process, called RIDA (regulatory inactivation of DnaA), ATP-bound DnaA transiently assembles into a complex consisting of Hda and the DNA-clamp, which promotes inter-AAA+ domain association between Hda and DnaA and stimulates hydrolysis of DnaA-bound ATP, producing inactive ADP-DnaA. Using a truncated DnaA mutant, we previously demonstrated that the DnaA N-terminal domain is involved in RIDA. However, the precise role of the N-terminal domain in RIDA has remained largely unclear. Here, we used an in vitro reconstituted system to demonstrate that the Asn-44 residue in the N-terminal domain of DnaA is crucial for RIDA but not for replication initiation. Moreover, an assay termed PDAX (pull-down after cross-linking) revealed an unstable interaction between a DnaA-N44A mutant and Hda. In vivo, this mutant exhibited an increase in the cellular level of ATP-bound DnaA. These results establish a model in which interaction between DnaA Asn-44 and Hda stabilizes the association between the AAA+ domains of DnaA and Hda to facilitate DnaA-ATP hydrolysis during RIDA. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

  15. Urinary urea nitrogen excretion during the hyperinsulinemic euglycemic clamp in type 1 diabetic patients and healthy subjects.

    PubMed

    Wohl, P; Krusinová, E; Klementová, M; Wohl, P; Kratochvílová, S; Pelikánová, T

    2008-01-01

    The hyperinsulinemic euglycemic clamp (HEC) combined with indirect calorimetry (IC) is used for estimation of insulin-stimulated substrate utilization. Calculations are based on urinary urea nitrogen excretion (UE), which is influenced by correct urine collection. The aims of our study were to improve the timing of urine collection during the clamp and to test the effect of insulin on UE in patients with type 1 diabetes (DM1; n=11) and healthy subjects (C; n=11). Urine samples were collected (a) over 24 h divided into 3-h periods and (b) before and during two-step clamp (1 and 10 mIU.kg(-1).min(-1); period 1 and period 2) combined with IC. The UE during the clamp was corrected for changes in urea pool size (UEc). There were no significant differences in 24-h UE between C and DM1 and no circadian variation in UE in either group. During the clamp, serum urea decreased significantly in both groups (p<0.01). Therefore, UEc was significantly lower as compared to UE not adjusted for changes in urea pool size both in C (p<0.001) and DM1 (p<0.001). While UE did not change during the clamp, UEc decreased significantly in both groups (p<0.01). UEc during the clamp was significantly higher in DM1 compared to C both in period 1 (p<0.05) and period 2 (p<0.01). The UE over 24 h and UEc during the clamp were statistically different in both C and DM1. We conclude that urine collection performed during the clamp with UE adjusted for changes in urea pool size is the most suitable technique for measuring substrate utilization during the clamp both in DM1 and C. Urine collections during the clamp cannot be replaced either by 24-h sampling (periods I-VII) or by a single 24-h urine collection. Attenuated insulin-induced decrease in UEc in DM1 implicates the impaired insulin effect on proteolysis.

  16. Efficacy of tranexamic acid plus drain-clamping to reduce blood loss in total knee arthroplasty: A meta-analysis.

    PubMed

    Zhang, Yan; Zhang, Jun-Wei; Wang, Bao-Hua

    2017-06-01

    Perioperative blood loss is still an unsolved problem in total knee arthroplasty (TKA). The efficacy of the preoperative use of tranexamic acid (TXA) plus drain-clamping to reduce blood loss in TKA has been debated. This meta-analysis aimed to illustrate the efficacy of TXA plus drain-clamping to reduce blood loss in patients who underwent a TKA. In February 2017, a systematic computer-based search was conducted in PubMed, EMBASE, Web of Science, the Cochrane Database of Systematic Reviews, and Google Scholar. Data from patients prepared for TKA in studies that compared TXA plus drain-clamping versus TXA alone, drain-clamping alone, or controls were retrieved. The primary endpoint was the need for transfusion. The secondary outcomes were total blood loss, blood loss in drainage, the decrease in hemoglobin, and the occurrence of deep venous thrombosis. After testing for publication bias and heterogeneity between studies, data were aggregated for random-effects models when necessary. Ultimately, 5 clinical studies with 618 patients (TXA plus drain-clamping group = 249, control group = 130, TXA-alone group = 60, and drain-clamping group = 179) were included. TXA plus drain-clamping could decrease the need for transfusion, total blood loss, blood loss in drainage, and the decrease in hemoglobin than could the control group, the TXA-alone group, and the drain-clamping group (P < .05). There was no significant difference between the occurrence of deep venous thrombosis between the included groups (P > .05). TXA plus drain-clamping can achieve the maximum effects of hemostasis in patients prepared for primary TKA. Because the number and the quality of the included studies were limited, more high-quality randomized controlled trials are needed to identify the optimal dose of TXA and the clamping hours in patients prepared for TKA.

  17. Efficacy of tranexamic acid plus drain-clamping to reduce blood loss in total knee arthroplasty

    PubMed Central

    Zhang, Yan; Zhang, Jun-Wei; Wang, Bao-Hua

    2017-01-01

    Abstract Background: Perioperative blood loss is still an unsolved problem in total knee arthroplasty (TKA). The efficacy of the preoperative use of tranexamic acid (TXA) plus drain-clamping to reduce blood loss in TKA has been debated. This meta-analysis aimed to illustrate the efficacy of TXA plus drain-clamping to reduce blood loss in patients who underwent a TKA. Methods: In February 2017, a systematic computer-based search was conducted in PubMed, EMBASE, Web of Science, the Cochrane Database of Systematic Reviews, and Google Scholar. Data from patients prepared for TKA in studies that compared TXA plus drain-clamping versus TXA alone, drain-clamping alone, or controls were retrieved. The primary endpoint was the need for transfusion. The secondary outcomes were total blood loss, blood loss in drainage, the decrease in hemoglobin, and the occurrence of deep venous thrombosis. After testing for publication bias and heterogeneity between studies, data were aggregated for random-effects models when necessary. Results: Ultimately, 5 clinical studies with 618 patients (TXA plus drain-clamping group = 249, control group = 130, TXA-alone group = 60, and drain-clamping group = 179) were included. TXA plus drain-clamping could decrease the need for transfusion, total blood loss, blood loss in drainage, and the decrease in hemoglobin than could the control group, the TXA-alone group, and the drain-clamping group (P < .05). There was no significant difference between the occurrence of deep venous thrombosis between the included groups (P > .05). Conclusions: TXA plus drain-clamping can achieve the maximum effects of hemostasis in patients prepared for primary TKA. Because the number and the quality of the included studies were limited, more high-quality randomized controlled trials are needed to identify the optimal dose of TXA and the clamping hours in patients prepared for TKA. PMID:28658157

  18. Properties of voltage-activated Na+ and K+ currents in mouse hippocampal glial cells in situ and after acute isolation from tissue slices.

    PubMed

    Steinhäuser, C; Kressin, K; Kuprijanova, E; Weber, M; Seifert, G

    1994-10-01

    In the present study, we were interested in a quantitative analysis of voltage-activated channels in a subpopulation of hippocampal glial cells, termed "complex" cells. The patch-clamp technique in the whole-cell mode was applied to identified cells in situ and to glial cells acutely isolated from tissue slices. The outward current was composed of two components: a sustained and a transient current. The transient K+ channel had electrophysiological and pharmacological properties resembling those of the channel through which the A-currents pass. In addition, this glial A-type current possessed a significant Ca2+ dependence. The current parameters determined in situ or in isolated cells corresponded well. Due to space clamp problems in situ, properties of voltage-dependent Na+ currents were only analysed in suspended glial cells. The tetrodotoxin (TTX) sensitivity and the stationary and kinetic characteristics of this current were similar to corresponding properties of hippocampal neurons. These quantitative data demonstrate that at an early postnatal stage of central nervous system maturation, glial cells in situ express a complex pattern of voltage-gated ion channels. The results are compared to findings in other preparations and the possible consequences of transmitter-mediated channel modulation in glial cells are discussed.

  19. High quality cord blood banking is feasible with delayed clamping practices. The eight-year experience and current status of the national Swedish Cord Blood Bank.

    PubMed

    Frändberg, Sofia; Waldner, Berit; Konar, Jan; Rydberg, Lennart; Fasth, Anders; Holgersson, Jan

    2016-09-01

    The National Swedish Cord Blood Bank (NS-CBB) is altruistic and publicly funded. Herein we describe the status of the bank and the impact of delayed versus early clamping on cell number and volume. Cord Blood Units (CBUs) were collected at two University Hospitals in Sweden. Collected volume and nucleated cell content (TNC) were investigated in 146 consecutive Cord Blood (CB) collections sampled during the first quarter of 2012 and in 162 consecutive CB collections done in the first quarter of 2013, before and after clamping practices were changed from immediate to late (60 s) clamping. NS-CBB now holds close to 5000 units whereof 30 % are from non-Caucasian or mixed origins. Delayed clamping had no major effect on collection efficiency. The volume collected was slightly reduced (mean difference, 8.1 ml; 95 % CI, 1.3-15.0 ml; p = 0.02), while cell recovery was not (p = 0.1). The proportion of CBUs that met initial total TNC banking criteria was 60 % using a TNC threshold of 12.5 × 10(8), and 47 % using a threshold of 15 × 10(8) for the early clamping group and 52 and 37 % in the late clamping group. Following implementation of delayed clamping practices at NS-CBB; close to 40 % of the collections in the late clamping group still met the high TNC banking threshold and were eligible for banking, implicating that that cord blood banking is feasible with delayed clamping practices.

  20. Quantifying Insulin Sensitivity and Entero-Insular Responsiveness to Hyper- and Hypoglycemia in Ferrets

    PubMed Central

    Sui, Hongshu; Yi, Yaling; Yao, Jianrong; Liang, Bo; Sun, Xingshen; Hu, Shanming; Uc, Aliye; Nelson, Deborah J.; Ode, Katie Larson; Philipson, Louis H.; Engelhardt, John F.; Norris, Andrew W.

    2014-01-01

    Ferrets are an important emerging model of cystic fibrosis related diabetes. However, there is little documented experience in the use of advanced techniques to quantify aspects of diabetes pathophysiology in the ferret. Glycemic clamps are the gold standard technique to assess both insulin sensitivity and insulin secretion in humans and animal models of diabetes. We therefore sought to develop techniques for glycemic clamps in ferrets. To assess insulin sensitivity, we performed euglycemic hyperinsulinemic clamps in 5–6 week old ferrets in the anesthetized and conscious states. To assess insulin secretion, we performed hyperglycemic clamps in conscious ferrets. To evaluate responsiveness of ferret islet and entero-insular hormones to low glucose, a portion of the hyperglycemic clamps were followed by a hypoglycemic clamp. The euglycemic hyperinsulinemic clamps demonstrated insulin responsiveness in ferrets similar to that previously observed in humans and rats. The anesthetic isoflurane induced marked insulin resistance, whereas lipid emulsion induced mild insulin resistance. In conscious ferrets, glucose appearance was largely suppressed at 4 mU/kg/min insulin infusion, whereas glucose disposal was progressively increased at 4 and 20 mU/kg/min insulin. Hyperglycemic clamp induced first phase insulin secretion. Hypoglycemia induced a rapid diminishment of insulin, as well as a rise in glucagon and pancreatic polypeptide levels. The incretins GLP-1 and GIP were affected minimally by hyperglycemic and hypoglycemic clamp. These techniques will prove useful in better defining the pathophysiology in ferrets with cystic fibrosis related diabetes. PMID:24594704

  1. A Comparison of the Performance and Application Differences Between Manual and Automated Patch-Clamp Techniques

    PubMed Central

    Yajuan, Xiao; Xin, Liang; Zhiyuan, Li

    2012-01-01

    The patch clamp technique is commonly used in electrophysiological experiments and offers direct insight into ion channel properties through the characterization of ion channel activity. This technique can be used to elucidate the interaction between a drug and a specific ion channel at different conformational states to understand the ion channel modulators’ mechanisms. The patch clamp technique is regarded as a gold standard for ion channel research; however, it suffers from low throughput and high personnel costs. In the last decade, the development of several automated electrophysiology platforms has greatly increased the screen throughput of whole cell electrophysiological recordings. New advancements in the automated patch clamp systems have aimed to provide high data quality, high content, and high throughput. However, due to the limitations noted above, automated patch clamp systems are not capable of replacing manual patch clamp systems in ion channel research. While automated patch clamp systems are useful for screening large amounts of compounds in cell lines that stably express high levels of ion channels, the manual patch clamp technique is still necessary for studying ion channel properties in some research areas and for specific cell types, including primary cells that have mixed cell types and differentiated cells that derive from induced pluripotent stem cells (iPSCs) or embryonic stem cells (ESCs). Therefore, further improvements in flexibility with regard to cell types and data quality will broaden the applications of the automated patch clamp systems in both academia and industry. PMID:23346269

  2. MATLAB implementation of a dynamic clamp with bandwidth >125 KHz capable of generating INa at 37°C

    PubMed Central

    Clausen, Chris; Valiunas, Virginijus; Brink, Peter R.; Cohen, Ira S.

    2012-01-01

    We describe the construction of a dynamic clamp with bandwidth >125 KHz that utilizes a high performance, yet low cost, standard home/office PC interfaced with a high-speed (16 bit) data acquisition module. High bandwidth is achieved by exploiting recently available software advances (code-generation technology, optimized real-time kernel). Dynamic-clamp programs are constructed using Simulink, a visual programming language. Blocks for computation of membrane currents are written in the high-level matlab language; no programming in C is required. The instrument can be used in single- or dual-cell configurations, with the capability to modify programs while experiments are in progress. We describe an algorithm for computing the fast transient Na+ current (INa) in real time, and test its accuracy and stability using rate constants appropriate for 37°C. We then construct a program capable of supplying three currents to a cell preparation: INa, the hyperpolarizing-activated inward pacemaker current (If), and an inward-rectifier K+ current (IK1). The program corrects for the IR drop due to electrode current flow, and also records all voltages and currents. We tested this program on dual patch-clamped HEK293 cells where the dynamic clamp controls a current-clamp amplifier and a voltage-clamp amplifier controls membrane potential, and current-clamped HEK293 cells where the dynamic clamp produces spontaneous pacing behavior exhibiting Na+ spikes in otherwise passive cells. PMID:23224681

  3. Quantifying insulin sensitivity and entero-insular responsiveness to hyper- and hypoglycemia in ferrets.

    PubMed

    Sui, Hongshu; Yi, Yaling; Yao, Jianrong; Liang, Bo; Sun, Xingshen; Hu, Shanming; Uc, Aliye; Nelson, Deborah J; Ode, Katie Larson; Philipson, Louis H; Engelhardt, John F; Norris, Andrew W

    2014-01-01

    Ferrets are an important emerging model of cystic fibrosis related diabetes. However, there is little documented experience in the use of advanced techniques to quantify aspects of diabetes pathophysiology in the ferret. Glycemic clamps are the gold standard technique to assess both insulin sensitivity and insulin secretion in humans and animal models of diabetes. We therefore sought to develop techniques for glycemic clamps in ferrets. To assess insulin sensitivity, we performed euglycemic hyperinsulinemic clamps in 5-6 week old ferrets in the anesthetized and conscious states. To assess insulin secretion, we performed hyperglycemic clamps in conscious ferrets. To evaluate responsiveness of ferret islet and entero-insular hormones to low glucose, a portion of the hyperglycemic clamps were followed by a hypoglycemic clamp. The euglycemic hyperinsulinemic clamps demonstrated insulin responsiveness in ferrets similar to that previously observed in humans and rats. The anesthetic isoflurane induced marked insulin resistance, whereas lipid emulsion induced mild insulin resistance. In conscious ferrets, glucose appearance was largely suppressed at 4 mU/kg/min insulin infusion, whereas glucose disposal was progressively increased at 4 and 20 mU/kg/min insulin. Hyperglycemic clamp induced first phase insulin secretion. Hypoglycemia induced a rapid diminishment of insulin, as well as a rise in glucagon and pancreatic polypeptide levels. The incretins GLP-1 and GIP were affected minimally by hyperglycemic and hypoglycemic clamp. These techniques will prove useful in better defining the pathophysiology in ferrets with cystic fibrosis related diabetes.

  4. Spectral infrared hemispherical reflectance measurements for LDEF tray clamps

    NASA Technical Reports Server (NTRS)

    Cromwell, B. K.; Shepherd, S. D.; Pender, C. W.; Wood, B. E.

    1993-01-01

    Infrared hemispherical reflectance measurements that were made on 58 chromic acid anodized tray clamps from LDEF are described. The measurements were made using a hemiellipsoidal mirror reflectometer with interferometer for wavelengths between 2-15 microns. The tray clamps investigated were from locations about the entire spacecraft and provided the opportunity for comparing the effects of atomic oxygen at each location. Results indicate there was essentially no dependence on atomic oxygen fluence for the surfaces studied, but there did appear to be a slight dependence on solar radiation exposure. The reflectances of the front sides of the tray clamps consistently were slightly higher than for the protected rear tray clamp surfaces.

  5. Advanced motor driven clamped borehole seismic receiver

    DOEpatents

    Engler, B.P.; Sleefe, G.E.; Striker, R.P.

    1993-02-23

    A borehole seismic tool is described including a borehole clamp which only moves perpendicular to the borehole. The clamp is driven by an electric motor, via a right angle drive. When used as a seismic receiver, the tool has a three part housing, two of which are hermetically sealed. Accelerometers or geophones are mounted in one hermetically sealed part, the electric motor in the other hermetically sealed part, and the clamp and right angle drive in the third part. Preferably the tool includes cable connectors at both ends. Optionally a shear plate can be added to the clamp to extend the range of the tool.

  6. DnaA protein DNA-binding domain binds to Hda protein to promote inter-AAA+ domain interaction involved in regulatory inactivation of DnaA.

    PubMed

    Keyamura, Kenji; Katayama, Tsutomu

    2011-08-19

    Chromosomal replication is initiated from the replication origin oriC in Escherichia coli by the active ATP-bound form of DnaA protein. The regulatory inactivation of DnaA (RIDA) system, a complex of the ADP-bound Hda and the DNA-loaded replicase clamp, represses extra initiations by facilitating DnaA-bound ATP hydrolysis, yielding the inactive ADP-bound form of DnaA. However, the mechanisms involved in promoting the DnaA-Hda interaction have not been determined except for the involvement of an interaction between the AAA+ domains of the two. This study revealed that DnaA Leu-422 and Pro-423 residues within DnaA domain IV, including a typical DNA-binding HTH motif, are specifically required for RIDA-dependent ATP hydrolysis in vitro and that these residues support efficient interaction with the DNA-loaded clamp·Hda complex and with Hda in vitro. Consistently, substitutions of these residues caused accumulation of ATP-bound DnaA in vivo and oriC-dependent inhibition of cell growth. Leu-422 plays a more important role in these activities than Pro-423. By contrast, neither of these residues is crucial for DNA replication from oriC, although they are highly conserved in DnaA orthologues. Structural analysis of a DnaA·Hda complex model suggested that these residues make contact with residues in the vicinity of the Hda AAA+ sensor I that participates in formation of a nucleotide-interacting surface. Together, the results show that functional DnaA-Hda interactions require a second interaction site within DnaA domain IV in addition to the AAA+ domain and suggest that these interactions are crucial for the formation of RIDA complexes that are active for DnaA-ATP hydrolysis.

  7. DnaA Protein DNA-binding Domain Binds to Hda Protein to Promote Inter-AAA+ Domain Interaction Involved in Regulatory Inactivation of DnaA*

    PubMed Central

    Keyamura, Kenji; Katayama, Tsutomu

    2011-01-01

    Chromosomal replication is initiated from the replication origin oriC in Escherichia coli by the active ATP-bound form of DnaA protein. The regulatory inactivation of DnaA (RIDA) system, a complex of the ADP-bound Hda and the DNA-loaded replicase clamp, represses extra initiations by facilitating DnaA-bound ATP hydrolysis, yielding the inactive ADP-bound form of DnaA. However, the mechanisms involved in promoting the DnaA-Hda interaction have not been determined except for the involvement of an interaction between the AAA+ domains of the two. This study revealed that DnaA Leu-422 and Pro-423 residues within DnaA domain IV, including a typical DNA-binding HTH motif, are specifically required for RIDA-dependent ATP hydrolysis in vitro and that these residues support efficient interaction with the DNA-loaded clamp·Hda complex and with Hda in vitro. Consistently, substitutions of these residues caused accumulation of ATP-bound DnaA in vivo and oriC-dependent inhibition of cell growth. Leu-422 plays a more important role in these activities than Pro-423. By contrast, neither of these residues is crucial for DNA replication from oriC, although they are highly conserved in DnaA orthologues. Structural analysis of a DnaA·Hda complex model suggested that these residues make contact with residues in the vicinity of the Hda AAA+ sensor I that participates in formation of a nucleotide-interacting surface. Together, the results show that functional DnaA-Hda interactions require a second interaction site within DnaA domain IV in addition to the AAA+ domain and suggest that these interactions are crucial for the formation of RIDA complexes that are active for DnaA-ATP hydrolysis. PMID:21708944

  8. Improvement of DGGE analysis by modifications of PCR protocols for analysis of microbial community members with low abundance.

    PubMed

    Wang, Yong-Feng; Zhang, Fang-Qiu; Gu, Ji-Dong

    2014-06-01

    Denaturing gradient gel electrophoresis (DGGE) is a powerful technique to reveal the community structures and composition of microorganisms in complex natural environments and samples. However, positive and reproducible polymerase chain reaction (PCR) products, which are difficult to acquire for some specific samples due to low abundance of the target microorganisms, significantly impair the effective applications of DGGE. Thus, nested PCR is often introduced to generate positive PCR products from the complex samples, but one problem is also introduced: The total number of thermocycling in nested PCR is usually unacceptably high, which results in skewed community structures by generation of random or mismatched PCR products on the DGGE gel, and this was demonstrated in this study. Furthermore, nested PCR could not resolve the uneven representative issue with PCR products of complex samples with unequal richness of microbial population. In order to solve the two problems in nested PCR, the general protocol was modified and improved in this study. Firstly, a general PCR procedure was used to amplify the target genes with the PCR primers without any guanine cytosine (GC) clamp, and then, the resultant PCR products were purified and diluted to 0.01 μg ml(-1). Subsequently, the diluted PCR products were utilized as templates to amplify again with the same PCR primers with the GC clamp for 17 cycles, and the products were finally subjected to DGGE analysis. We demonstrated that this is a much more reliable approach to obtain a high quality DGGE profile with high reproducibility. Thus, we recommend the adoption of this improved protocol in analyzing microorganisms of low abundance in complex samples when applying the DGGE fingerprinting technique to avoid biased results.

  9. Sperm Patch-Clamp

    PubMed Central

    Lishko, Polina; Clapham, David E.; Navarro, Betsy; Kirichok, Yuriy

    2014-01-01

    Sperm intracellular pH and calcium concentration ([Ca2+]i) are two central factors that control sperm activity within the female reproductive tract. As such, the ion channels of the sperm plasma membrane that alter intracellular sperm [Ca2+] and pH play important roles in sperm physiology and the process of fertilization. Indeed, sperm ion channels regulate sperm motility, control sperm chemotaxis toward the egg in some species, and may trigger the acrosome reaction. Until recently, our understanding of these important molecules was rudimentary due to the inability to patch-clamp spermatozoa and directly record the activity of these ion channels under voltage clamp. Recently, we overcame this technical barrier and developed a method for reproducible application of the patch-clamp technique to mouse and human spermatozoa. This chapter covers important aspects of application of the patch-clamp technique to spermatozoa, such as selection of the electrophysiological equipment, isolation of spermatozoa for patch-clamp experiments, formation of the gigaohm seal with spermatozoa, and transition into the whole-cell mode of recording. We also discuss potential pitfalls in application of the patch-clamp technique to flagellar ion channels. PMID:23522465

  10. Acute exercise alters skeletal muscle mitochondrial respiration and H2O2 emission in response to hyperinsulinemic-euglycemic clamp in middle-aged obese men

    PubMed Central

    Trewin, Adam J.; Levinger, Itamar; Parker, Lewan; Shaw, Christopher S.; Serpiello, Fabio R.; Anderson, Mitchell J.; McConell, Glenn K.; Hare, David L.

    2017-01-01

    Obesity, sedentary lifestyle and aging are associated with mitochondrial dysfunction and impaired insulin sensitivity. Acute exercise increases insulin sensitivity in skeletal muscle; however, whether mitochondria are involved in these processes remains unclear. The aim of this study was to investigate the effects of insulin stimulation at rest and after acute exercise on skeletal muscle mitochondrial respiratory function (JO2) and hydrogen peroxide emission (JH2O2), and the associations with insulin sensitivity in obese, sedentary men. Nine men (means ± SD: 57 ± 6 years; BMI 33 ± 5 kg.m2) underwent hyperinsulinemic-euglycemic clamps in two separate trials 1–3 weeks apart: one under resting conditions, and another 1 hour after high-intensity exercise (4x4 min cycling at 95% HRpeak). Muscle biopsies were obtained at baseline, and pre/post clamp to measure JO2 with high-resolution respirometry and JH2O2 via Amplex UltraRed from permeabilized fibers. Post-exercise, both JO2 and JH2O2 during ADP stimulated state-3/OXPHOS respiration were lower compared to baseline (P<0.05), but not after subsequent insulin stimulation. JH2O2 was lower post-exercise and after subsequent insulin stimulation compared to insulin stimulation in the rest trial during succinate supported state-4/leak respiration (P<0.05). In contrast, JH2O2 increased during complex-I supported leak respiration with insulin after exercise compared with resting conditions (P<0.05). Resting insulin sensitivity and JH2O2 during complex-I leak respiration were positively correlated (r = 0.77, P<0.05). We conclude that in obese, older and sedentary men, acute exercise modifies skeletal muscle mitochondrial respiration and H2O2 emission responses to hyperinsulinemia in a respiratory state-specific manner, which may have implications for metabolic diseases involving insulin resistance. PMID:29161316

  11. Zero ischemia anatomical partial nephrectomy: a novel approach.

    PubMed

    Gill, Inderbir S; Patil, Mukul B; Abreu, Andre Luis de Castro; Ng, Casey; Cai, Jie; Berger, Andre; Eisenberg, Manuel S; Nakamoto, Masahiko; Ukimura, Osamu; Goh, Alvin C; Thangathurai, Duraiyah; Aron, Monish; Desai, Mihir M

    2012-03-01

    We present a novel concept of zero ischemia anatomical robotic and laparoscopic partial nephrectomy. Our technique primarily involves anatomical vascular microdissection and preemptive control of tumor specific, tertiary or higher order renal arterial branch(es) using neurosurgical aneurysm micro-bulldog clamps. In 58 consecutive patients the majority (70%) had anatomically complex tumors including central (67%), hilar (26%), completely intrarenal (23%), pT1b (18%) and solitary kidney (7%). Data were prospectively collected and analyzed from an institutional review board approved database. Of 58 cases undergoing zero ischemia robotic (15) or laparoscopic (43) partial nephrectomy, 57 (98%) were completed without hilar clamping. Mean tumor size was 3.2 cm, mean ± SD R.E.N.A.L. score 7.0 ± 1.9, C-index 2.9 ± 2.4, operative time 4.4 hours, blood loss 206 cc and hospital stay 3.9 days. There were no intraoperative complications. Postoperative complications (22.8%) were low grade (Clavien grade 1 to 2) in 19.3% and high grade (Clavien grade 3 to 5) in 3.5%. All patients had negative cancer surgical margins (100%). Mean absolute and percent change in preoperative vs 4-month postoperative serum creatinine (0.2 mg/dl, 18%), estimated glomerular filtration rate (-11.4 ml/minute/1.73 m(2), 13%), and ipsilateral kidney function on radionuclide scanning at 6 months (-10%) correlated with mean percent kidney excised intraoperatively (18%). Although 21% of patients received a perioperative blood transfusion, no patient had acute or delayed renal hemorrhage, or lost a kidney. The concept of zero ischemia robotic and laparoscopic partial nephrectomy is presented. This anatomical vascular microdissection of the artery first and then tumor allows even complex tumors to be excised without hilar clamping. Global surgical renal ischemia is unnecessary for the majority of patients undergoing robotic and laparoscopic partial nephrectomy at our institution. Copyright © 2012 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

  12. USSR Report, International Affairs

    DTIC Science & Technology

    1987-02-11

    shown that the cycle "A New Development—Assimilation in Production " has accelerated to practically twice its original speed thanks to these forms of... new production capacities to manufacture hydrodynamic transmissions for scrapers and loaders, and in doing so, will be assisted by a partner—the...creation of new forms of production through joint efforts. Of primary importance in achieving these new forms are mobile and flexible contacts that are in

  13. Conceptual Design of a Robotic Loader System for Remote Missile Launchers.

    DTIC Science & Technology

    1985-09-01

    artifcial intelligence were sur- veyed in order to assess their space applicability and to identify areas which can be developed/adapted to European...such data bases as NTIS and COMPENDEX. The second computer aided search was done through the U. S. Army information services at Redstone Arsenal...Lockheed Corporation. The first DIALOG data base explored was NTIS (National Technical Information Services, U.S. Dept. of Commerce), which contains

  14. Family History of Breast Cancer as a Determinant of the Risk of Developing Endometrial and Ovarian Cancers: A Nationwide Cohort Study

    DTIC Science & Technology

    2002-01-01

    TS, Manley SA, Olopade OI, Cummings S, Garber JE, Bernhardt B, Antman K, Russo D, Wood ME, Mullineau L, Isaacs C, Peshkin B, Buys S, Venne V, Rowley... Antman K, Russo D, Wood ME, Mullineau L, Isaacs C, Peshkin B, Buys S, Venne V, Rowley PT, Loader S, Offit K, Robson M, Hampel H, Brener D, Winer EP

  15. Final Environmental Assessment for a Proposed Pararescue and Combat Rescue Officer Training Campus at Kirtland Air Force Base

    DTIC Science & Technology

    2006-09-01

    tank with adjacent training structure and hoist for inflatable boat training. An indoor climbing wall, a weight/fitness room, and staff and student...climbing; rappelling/litter evacuation; Tyrolean traverses; suspended harness parachuting emergency procedures; aircraft hoist infiltration...characteristics in common. Bulldozers, backhoes, and front-end loaders would be on site throughout periods of excavation and/or site preparation. Dump

  16. Expedient Repair of Structural Facilities (ERSF). Volume 1. ERSF System Development

    DTIC Science & Technology

    1992-01-01

    slabs, will require a different vehicle for transportation; for example a flatbed truck. Specialized equipment, such as the shotcrete machine described in...1) glued, laminated timber (glulam) column insertion and (2) mechanical shoring jack insertion. Both systems were ranked essentially equally after...gage wire mesh; - (optional) 100 rebar ties; and - in some cases, a bulldozer, front-end loader, and flatbed truck. c. Training Since a new piece of

  17. Interpretive computer simulator for the NASA Standard Spacecraft Computer-2 (NSSC-2)

    NASA Technical Reports Server (NTRS)

    Smith, R. S.; Noland, M. S.

    1979-01-01

    An Interpretive Computer Simulator (ICS) for the NASA Standard Spacecraft Computer-II (NSSC-II) was developed as a code verification and testing tool for the Annular Suspension and Pointing System (ASPS) project. The simulator is written in the higher level language PASCAL and implented on the CDC CYBER series computer system. It is supported by a metal assembler, a linkage loader for the NSSC-II, and a utility library to meet the application requirements. The architectural design of the NSSC-II is that of an IBM System/360 (S/360) and supports all but four instructions of the S/360 standard instruction set. The structural design of the ICS is described with emphasis on the design differences between it and the NSSC-II hardware. The program flow is diagrammed, with the function of each procedure being defined; the instruction implementation is discussed in broad terms; and the instruction timings used in the ICS are listed. An example of the steps required to process an assembly level language program on the ICS is included. The example illustrates the control cards necessary to assemble, load, and execute assembly language code; the sample program to to be executed; the executable load module produced by the loader; and the resulting output produced by the ICS.

  18. Assessing combined exposures of whole-body vibration and awkward posture--further results from application of a simultaneous field measurement methodology.

    PubMed

    Raffler, Nastaran; Hermanns, Ingo; Sayn, Detlef; Göres, Benno; Ellegast, Rolf; Rissler, Jörg

    2010-01-01

    The drivers of ten vehicles (tram, helicopter, saloon car, van, forklift, two mobile excavators, wheel loader, tractor, elevating platform truck) were studied with regard to the combined exposures of whole-body vibration and awkward posture during occupational tasks. Seven degrees of freedom (DOFs), or body angles, were recorded as a function of time by means of the CUELA measuring system (Computer-assisted registration and long-term analysis of musculoskeletal workloads) for the purpose of posture assessment. The vibrational exposure is expressed as the vector sum of the frequency-weighted accelerations in the three Cartesian coordinates; these were recorded simultaneously with the posture measurement. Based upon the percentage of working time spent under different workloads, a scheme is proposed for classification of the two exposures into three categories. In addition, a risk of adverse health effects classified as low, possible or high can be assigned to the combination of the two exposures. With regard to posture, the most severe exposure was measured for the drivers of the wheel loader and for the tractor driver, whereas the lowest exposure was measured for the helicopter pilots and van drivers. With regard to the combination of whole-body and posture exposures, the tractor driver and the elevating platform truck driver exhibited the highest workloads.

  19. Open-access microfluidic patch-clamp array with raised lateral cell trapping sites.

    PubMed

    Lau, Adrian Y; Hung, Paul J; Wu, Angela R; Lee, Luke P

    2006-12-01

    A novel open-access microfluidic patch-clamp array chip with lateral cell trapping sites raised above the bottom plane of the chip was developed by combining both a microscale soft-lithography and a macroscale polymer fabrication method. This paper demonstrates the capability of using such an open-access fluidic system for patch-clamp measurements. The surface of the open-access patch-clamp sites prepared by the macroscale hole patterning method of soft-state elastic polydimethylsiloxane (PDMS) is examined; the seal resistances are characterized and correlated with the aperture dimensions. Whole cell patch-clamp measurements are carried out with CHO cells expressing Kv2.1 ion channels. Kv2.1 ion channel blocker (TEA) dosage response is characterized and the binding activity is examined. The results demonstrate that the system is capable of performing whole cell measurements and drug profiling in a more efficient manner than the traditional patch-clamp set-up.

  20. RNA polymerase pausing and nascent RNA structure formation are linked through clamp domain movement

    PubMed Central

    Hein, Pyae P.; Kolb, Kellie E.; Windgassen, Tricia; Bellecourt, Michael J.; Darst, Seth A.; Mooney, Rachel A.; Landick, Robert

    2014-01-01

    The rates of RNA synthesis and nascent RNA folding into biologically active structures are linked via pausing by RNA polymerase (RNAP). Structures that form within the RNA exit channel can increase pausing by interacting with bacterial RNAP or decrease pausing by preventing backtracking. Conversely, pausing is required for proper folding of some RNAs. Opening of the RNAP clamp domain is proposed to mediate some effects of nascent RNA structures. However, the connections among RNA structure formation, clamp movement, and catalytic activity remain uncertain. We assayed exit-channel structure formation in Escherichia coli RNAP together with disulfide crosslinks that favor closed or open clamp conformations and found that clamp position directly influences RNA structure formation and catalytic activity. We report that exit-channel RNA structures slow pause escape by favoring clamp opening and through interactions with the flap that slow translocation. PMID:25108353

  1. Cell-Detection Technique for Automated Patch Clamping

    NASA Technical Reports Server (NTRS)

    McDowell, Mark; Gray, Elizabeth

    2008-01-01

    A unique and customizable machinevision and image-data-processing technique has been developed for use in automated identification of cells that are optimal for patch clamping. [Patch clamping (in which patch electrodes are pressed against cell membranes) is an electrophysiological technique widely applied for the study of ion channels, and of membrane proteins that regulate the flow of ions across the membranes. Patch clamping is used in many biological research fields such as neurobiology, pharmacology, and molecular biology.] While there exist several hardware techniques for automated patch clamping of cells, very few of those techniques incorporate machine vision for locating cells that are ideal subjects for patch clamping. In contrast, the present technique is embodied in a machine-vision algorithm that, in practical application, enables the user to identify good and bad cells for patch clamping in an image captured by a charge-coupled-device (CCD) camera attached to a microscope, within a processing time of one second. Hence, the present technique can save time, thereby increasing efficiency and reducing cost. The present technique involves the utilization of cell-feature metrics to accurately make decisions on the degree to which individual cells are "good" or "bad" candidates for patch clamping. These metrics include position coordinates (x,y) in the image plane, major-axis length, minor-axis length, area, elongation, roundness, smoothness, angle of orientation, and degree of inclusion in the field of view. The present technique does not require any special hardware beyond commercially available, off-the-shelf patch-clamping hardware: A standard patchclamping microscope system with an attached CCD camera, a personal computer with an imagedata- processing board, and some experience in utilizing imagedata- processing software are all that are needed. A cell image is first captured by the microscope CCD camera and image-data-processing board, then the image data are analyzed by software that implements the present machine-vision technique. This analysis results in the identification of cells that are "good" candidates for patch clamping (see figure). Once a "good" cell is identified, a patch clamp can be effected by an automated patchclamping apparatus or by a human operator. This technique has been shown to enable reliable identification of "good" and "bad" candidate cells for patch clamping. The ultimate goal in further development of this technique is to combine artificial-intelligence processing with instrumentation and controls in order to produce a complete "turnkey" automated patch-clamping system capable of accurately and reliably patch clamping cells with a minimum intervention by a human operator. Moreover, this technique can be adapted to virtually any cellular-analysis procedure that includes repetitive operation of microscope hardware by a human.

  2. Lotus birth, a holistic approach on physiological cord clamping.

    PubMed

    Zinsser, Laura A

    2018-04-01

    The positive effects of delayed cord clamping (DCC) has been extensively researched. DCC means: waiting at least one minute after birth before clamping and cutting the cord or till the pulsation has stopped. With physiological clamping and cutting (PCC) the clamping and cutting can happen at the earliest after the pulsation has stopped. With a Lotus birth, no clamping and cutting of the cord is done. A woman called Clair Lotus Day imitated the holistic approach of PCC from an anthropoid ape in 1974. The chimpanzee did not separate the placenta from the newborn. The aim of this case report is to discuss and learn a different approach in the third stage of labour. Three cases of Lotus birth by human beings were observed. All three women gave birth in an out-of-hospital setting and had ambulant postnatal care. The placenta was washed, salted and herbs were put on 2-3h post partum. The placenta was wrapped in something that absorbs the moisture. The salting was repeated with a degreasing frequency depending on moistness of the placenta. On life day six all three Lotus babies experiences a natural separation of the cord. All three Lotus birth cases were unproblematic, no special incidence occurred. One should differentiate between early cord clamping (ECC), delayed cord clamping (DCC) and physiological cord clamping (PCC). Lotus birth might lead to an optimisation of the bonding and attachment. Research is needed in the areas of both PCC and Lotus birth. Copyright © 2017 Australian College of Midwives. Published by Elsevier Ltd. All rights reserved.

  3. A novel automotive headlight system based on digital micro-mirror devices and diffractive optical elements

    NASA Astrophysics Data System (ADS)

    Su, Ping; Song, Yuming; Ma, Jianshe

    2018-01-01

    The DMD (Digital Micro-mirror Device) has the advantages of high refresh rate and high diffraction efficiency, and these make it become an ideal loader of multiple modes illumination. DOEs (Diffractive Optical Element) have the advantages of high degree of freedom, light weight, easy to copy, low cost etc., and can be used to reduce the weight, complexity, cost of optical system. A novel automotive headlamp system using DMD as the light distribution element and a DOE as the light field modulation device is proposed in this paper. The pure phase DOE is obtained by the GS algorithm using Rayleigh-Sommerfeld diffraction integral model. Based on the standard automotive headlamp light intensity distribution in the target plane, the amplitude distribution of DMD is obtained by numerical simulation, and the grayscale diagram loaded on the DMD can be obtained accordingly. Finally, according to simulation result, the light intensity distribution in the target plane is proportional to the national standard, hence verifies the validity of the novel system. The novel illumination system proposed in this paper provides a reliable hardware platform for the intelligent headlamps.

  4. Independent mechanisms recruit the cohesin loader protein NIPBL to sites of DNA damage.

    PubMed

    Bot, Christopher; Pfeiffer, Annika; Giordano, Fosco; Manjeera, Dharani E; Dantuma, Nico P; Ström, Lena

    2017-03-15

    NIPBL is required to load the cohesin complex on to DNA. While the canonical role of cohesin is to couple replicated sister chromatids together until the onset of mitosis, it also promotes tolerance to DNA damage. Here, we show that NIPBL is recruited to DNA damage throughout the cell cycle via independent mechanisms, influenced by type of damage. First, the heterochromatin protein HP1γ (also known as CBX3) recruits NIPBL to DNA double-strand breaks (DSBs) through the corresponding HP1-binding motif within the N-terminus. By contrast, the C-terminal HEAT repeat domain is unable to recruit NIPBL to DSBs but independently targets NIPBL to laser microirradiation-induced DNA damage. Each mechanism is dependent on the RNF8 and RNF168 ubiquitylation pathway, while the recruitment of the HEAT repeat domain requires further ATM or ATR activity. Thus, NIPBL has evolved a sophisticated response to damaged DNA that is influenced by the form of damage, suggesting a highly dynamic role for NIPBL in maintaining genomic stability. © 2017. Published by The Company of Biologists Ltd.

  5. Mechanical coupling for a rotor shaft assembly of dissimilar materials

    DOEpatents

    Shi, Jun [Glastonbury, CT; Bombara, David [New Hartford, CT; Green, Kevin E [Broad Brook, CT; Bird, Connic [Rocky Hill, CT; Holowczak, John [South Windsor, CT

    2009-05-05

    A mechanical coupling for coupling a ceramic disc member to a metallic shaft includes a first wedge clamp and a second wedge clamp. A fastener engages a threaded end of a tie-bolt to sandwich the ceramic disc between the wedge clamps. An axial spring is positioned between the fastener and the second wedge clamp to apply an axial preload along the longitudinal axis. Another coupling utilizes a rotor shaft end of a metallic rotor shaft as one wedge clamp. Still another coupling includes a solid ceramic rotor disc with a multiple of tie-bolts radially displaced from the longitudinal axis to exert the preload on the solid ceramic rotor disc.

  6. Laser beam guard clamps

    DOEpatents

    Dickson, Richard K.

    2010-09-07

    A quick insert and release laser beam guard panel clamping apparatus having a base plate mountable on an optical table, a first jaw affixed to the base plate, and a spring-loaded second jaw slidably carried by the base plate to exert a clamping force. The first and second jaws each having a face acutely angled relative to the other face to form a V-shaped, open channel mouth, which enables wedge-action jaw separation by and subsequent clamping of a laser beam guard panel inserted through the open channel mouth. Preferably, the clamping apparatus also includes a support structure having an open slot aperture which is positioned over and parallel with the open channel mouth.

  7. Launch Lock Assemblies with Reduced Preload and Spacecraft Isolation Systems Including the Same

    NASA Technical Reports Server (NTRS)

    Barber, Tim Daniel (Inventor); Young, Ken (Inventor); Hindle, Timothy (Inventor)

    2016-01-01

    Launch lock assemblies with reduced preload are provided. The launch lock assembly comprises first and second mount pieces, a releasable clamp device, and a pair of retracting assemblies. Each retracting assembly comprises a pair of toothed members having interacting toothed surfaces. The releasable clamp device normally maintains the first and second mount pieces in clamped engagement. When the releasable clamp device is actuated, the first and second mount pieces are released from clamped engagement and one toothed member of each retracting assembly moves in an opposite direction relative to the other one toothed member of the other retracting assembly to define an axial gap on each side of the first mount piece.

  8. Internal V-Band Clamp

    DOEpatents

    Vaughn, Mark R.; Hafenrichter, Everett S.; Chapa, Agapito C.; Harris, Steven M.; Martinez, Marcus J.; Baty, Roy S.

    2006-02-28

    A system for clamping two tubular members together in an end-to-end relationship uses a split ring with a V-shaped outer rim that can engage a clamping surface on each member. The split ring has a relaxed closed state where the ends of the ring are adjacent and the outside diameter of the split ring is less than the minimum inside diameter of the members at their ends. The members are clamped when the split ring is spread into an elastically stretched position where the ring rim is pressed tightly against the interior surfaces of the members. Mechanisms are provided for removing the spreader so the split ring will return to the relaxed state, releasing the clamped members.

  9. Pipe support

    DOEpatents

    Pollono, Louis P.

    1979-01-01

    A pipe support for high temperature, thin-walled piping runs such as those used in nuclear systems. A section of the pipe to be supported is encircled by a tubular inner member comprised of two walls with an annular space therebetween. Compacted load-bearing thermal insulation is encapsulated within the annular space, and the inner member is clamped to the pipe by a constant clamping force split-ring clamp. The clamp may be connected to pipe hangers which provide desired support for the pipe.

  10. Self-aligning fixture used in lathe chuck jaw refacing

    NASA Technical Reports Server (NTRS)

    Linn, C. C.

    1965-01-01

    Self-aligning tool positions and rigidly holds lathe chuck jaws for refacing and truing of the clamping surface. The jaws clamp the fixture in the manner of clamping a workpiece. The fixture can be modified to accommodate four-jawed checks.

  11. The optimal protocol to reduce blood loss and blood transfusion after unilateral total knee replacement: Low-dose IA-TXA plus 30-min drain clamping versus drainage clamping for the first 3 h without IA-TXA.

    PubMed

    Park, Joo Hyun; Choi, Sung Wook; Shin, Eun Ho; Park, Myung Hoon; Kim, Myung Ku

    2017-01-01

    Although intraarticular tranexamic acid (IA-TXA) administration or drainage clamping are popular methods used to reduce blood loss after total knee replacement (TKR), the protocol remains controversial. We aimed (1) to establish new protocols through investigating whether two methods, that is, low-dose (500 mg) IA-TXA plus 30-min drain clamping and drainage clamping for the first 3 h without IA-TXA, can reduce blood loss and blood transfusion after unilateral TKR and (2) to make recommendations related to clinical application. This study, conducted from September 2014 to June 2016 related to enrolled 95 patients with primary osteoarthritis who were to have a unilateral cemented TKR, was nonrandomized and retrospective. In group A, the drain was released following tourniquet deflation. In group B, 500-mg TXA was injected into the knee joint via a drain tube after fascia closure and the drain was clamped for the first 30 min to prevent leakage. In group C, the drain was clamped for the first 3-h postoperation. Demographic characteristics and clinical data were collected, including the levels of hematocrit (Hct), the total blood loss (TBL), drained blood volume (BV), the amount of blood transfused, and any complications that developed. We found a significantly lower postoperative TBL, drained BV, decreasing Hct level, and less transfused BV in the IA-TXA injection group (group B) and the 3-h drainage clamping group (group C) compared to the conventional negative drainage group (group A; p < 0.001). There was no significant difference between groups B and C ( p = 0.99). The drainage clamping method can be safer than IA-TXA administration in patients with risk factor of venous thromboembolic complication. Furthermore, the IA-TXA administration can be more optimal than drainage clamping in patients with high bleeding tendency or lateral retinacular release during TKR, who would be concerned about postoperative wound complication.

  12. Temporary clamping of external carotid artery in convexity, parasagittal and temporal base meningioma.

    PubMed

    Yadav, Yad Ram; Parihar, Vijay; Agarwal, Moneet; Bhatele, Pushp Raj

    2012-01-01

    The management of intraoperative bleeding during removal of a large hyper vascular meningioma is crucial for safe and efficient surgery. Preoperative embolization of meningioma is the best way to reduce vascularity of meningiomas but this technique is not readily available, costly and has its own limitations. The study is aimed to evaluate the use of temporary clamping of external carotid artery to reduce blood loss and operating time during excision of large convexity, parasagittal or temporal base meningiomas. A prospective study of 115 consecutively operated meningiomas of size 5 cms or more were operated from January 2002 to December2010. Temporary clamping of external carotid artery was done in 61 while 51 cases were managed without clamping. There was significant reduction of blood loss, operative time and blood transfusion given in the temporary clipping group compared to non clipping group. There was stitch abscess in two patients each in clamping, and non clamping group. There was no scalp necrosis or mortality in any of the group. Temporary clamping of external carotid artery is a safe, simple and cost-effective alternative to embolization for the surgery of large meningiomas. This can be practiced at all the centers.

  13. Measuring true Young's modulus of a cantilevered nanowire: effect of clamping on resonance frequency.

    PubMed

    Qin, Qingquan; Xu, Feng; Cao, Yongqing; Ro, Paul I; Zhu, Yong

    2012-08-20

    The effect of clamping on resonance frequency and thus measured Young's modulus of nanowires (NWs) is systematically investigated via a combined experimental and simulation approach. ZnO NWs are used in this work as an example. The resonance tests are performed in situ inside a scanning electron microscope and the NWs are cantilevered on a tungsten probe by electron-beam-induced deposition (EBID) of hydrocarbon. EBID is repeated several times to deposit more hydrocarbons at the same location. The resonance frequency increases with the increasing clamp size until approaching that under the "fixed" boundary condition. The critical clamp size is identified as a function of NW diameter and NW Young's modulus. This work: 1) exemplifies the importance of considering the effect of clamping in measurements of Young's modulus using the resonance method, and 2) demonstrates that the true Young's modulus can be measured if the critical clamp size is reached. Design guidelines on the critical clamp size are provided. Such design guidelines can be extended to other one-dimensional nanostructures such as carbon nanotubes. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Peptide- and proton-driven allosteric clamps catalyze anthrax toxin translocation across membranes

    PubMed Central

    Das, Debasis; Krantz, Bryan A.

    2016-01-01

    Anthrax toxin is an intracellularly acting toxin in which sufficient information is available regarding the structure of its transmembrane channel, allowing for detailed investigation of models of translocation. Anthrax toxin, comprising three proteins—protective antigen (PA), lethal factor (LF), and edema factor—translocates large proteins across membranes. Here we show that the PA translocase channel has a transport function in which its catalytic active sites operate allosterically. We find that the phenylalanine clamp (ϕ-clamp), the known conductance bottleneck in the PA translocase, gates as either a more closed state or a more dilated state. Thermodynamically, the two channel states have >300-fold different binding affinities for an LF-derived peptide. The change in clamp thermodynamics requires distant α-clamp and ϕ-clamp sites. Clamp allostery and translocation are more optimal for LF peptides with uniform stereochemistry, where the least allosteric and least efficiently translocated peptide had a mixed stereochemistry. Overall, the kinetic results are in less agreement with an extended-chain Brownian ratchet model but, instead, are more consistent with an allosteric helix-compression model that is dependent also on substrate peptide coil-to-helix/helix-to-coil cooperativity. PMID:27506790

  15. Plasma Chamber Restraints in Ignitor and Relevant Disruption Analysis

    NASA Astrophysics Data System (ADS)

    Gasparotto, M.; Cucchiaro, A.; Capriccioli, A.; Celentano, G.; Rita, C.; Roccella, M.; Macco, B.; Micheli, I.; Ferrari, G.; Orlandi, S.; Coppi, B.

    2000-10-01

    The plasmas chamber (PC) of Ignitor is made of 12 D-shaped toroidal sectors of Inconel 625 welded together by automatic remote equipment. The thickness of the inboard wall is 17 mm while the middle and outboard walls are 26 mm thick. The PC is supported through the ports by the C-Clamp structure of the toroidal magnet. The main function of the PC supports is to resist the vertical and radial electromagnetic loads and to allow for free movement under thermal loads while providing electrical insulation from the C-Clamps and cryostat. The largest estimated loads are due to a Vertical Displacement Event (VDE) disruption that is followed by a thermal quench and then by the current quench. The vertical supports involve a connection of each radial port to the C-Clamp structure by a link system that withstands the calculated loads. The radial supports resist, with high stiffness, the centripetal and centrifugal forces. The end flange of each radial port is connected to the C-Clamp structure by a clamping sleeve device. The clamping sleeves are hydraulically operated to provide locking during discharge. The clamping sleeves of the radial support system have been validated by an appropriate series of tests.

  16. Quick action clamp

    NASA Technical Reports Server (NTRS)

    Calco, Frank S. (Inventor)

    1991-01-01

    A quick release toggle clamp that utilizes a spring that requires a deliberate positive action for disengagement is presented. The clamp has a sliding bolt that provides a latching mechanism. The bolt is moved by a handle that tends to remain in an engaged position while under tension.

  17. Rotary slot dog

    DOEpatents

    Cutburth, Ronald W.; Smauley, David A.

    1987-01-01

    A clamp or dog is disclosed which preferably comprises a slotted stepped cylindrical body which is inserted into a hole in a workpiece and then fastened to a base or fixture using a screw which is inserted through the slot. The stepped configuration provides an annular clamping surface which securely clamps the workpiece against the base or fixture. The slotted cylindrical configuration permits adjustment of the workpiece and retaining clamp in any direction, i.e., over 360.degree., relative to the mounting position of the screw in the base or fixture.

  18. Improved Intermittent-clamped Drainage in Lower Lumbar Internal Fixation: A Randomized Prospective Study

    PubMed Central

    Hao, Qing-Ying; Liu, Chu-Yin; Fu, Chan-Juan; Zhang, Xiao-Hua; Tan, Ming-Sheng

    2016-01-01

    Background: Continuous negative pressure drainage (CNPD) is widely used after lower lumbar internal fixation; however, it may cause tremendous blood loss and lead to postoperative hemorrhagic anemia. The present study explored the efficacy and safety of improved intermittent-clamped drainage (ICD) for lower lumbar internal fixation. Methods: This was a prospective study that included 156 patients with decompression of the spinal canal and internal fixation for the first time from January 2012 to December 2014. The patients were randomly divided into ICD group and CNPD group, and each group had 78 cases. A drainage tube was placed under the deep fascia in all patients within 10 min after the commencement of wound closure. The postoperative drainage amount at different time points, the hemoglobin level, and postoperative complications were recorded and compared between the two groups. Shapiro-Wilk test, independent samples t-test, and Mann-Whitney U-test were used in this study. Results: The drainage amount was significantly reduced in the ICD group, as compared with the CNPD group (Z = 10.74, P < 0.01). The mean total drainage amount (in ml) of the single-segment and two-segment procedures was significantly greater in the CNPD group than the ICD group (Z = 10.63 and 10.75, respectively; P < 0.01). For the adverse events, there was no significant difference in postoperative temperature, wound problem, and complications between the two groups. Conclusions: The present study showed a statistically significant reduction in postoperative drainage amount between ICD and CNPD groups, and ICD is an effective, convenient, and safe method for routine use in lower lumbar surgery. It is essential to focus on the effect of clamping drainage with long-segment surgical procedure and complex lumbar disease in the further investigation, as well as the effect of clamping on long-term functional outcomes. PMID:27900992

  19. Study of clamping-induced deformations on SR optics by means of LTP

    NASA Astrophysics Data System (ADS)

    Cocco, Daniele; Sostero, Giovanni; Zangrando, Marco

    2001-01-01

    With the advent of third generation synchrotron radiation sources, more flux was available for the experimentalist. At the same time, the request in term of spot dimension and energy resolution rapidly increased. For this reason, opticians try to design beamlines with higher and higher performances. To this end the shape of every optical component of a beamline is specified to have very tight constrains, because every small figure error produces a sudden reduction in terms of the overall performance. Nevertheless, the necessity to positioning and cooling the components implies the presence of a safe clamping system which unavoidably would modifies the shape of the component, causing possible reduction of resolving power or increasing the spot dimension. Thus it is not sufficient to measure accurately the slope or the profile of a mirror in laboratory before the mounting, but it is useful to test it also after this procedure. We, at ELETTRA, have measured by means of a modified version of the LTP II (Long Trace Profiler) several mirrors and gratings before and after their clamping, in order to estimate the effect of the holder on the final performances of the beamlines. Since our LTP II measures directly the local slope of the surface under test with a repeatability better than 0.02 arcsec on a 1 meter long optical surface, it is very easy to single out any small distortion of the tangential profile introduced by the mounting system. Different kinds of supports for both small and large optical elements, were taken into consideration and the effect of the deformation induced by them on the beamline performance was simulated and will be presented here together with the results of each measurements. The results give us a way to select properly the kind of clamping and invite the opticians to try to take into consideration also this effect before designing a complex beamline.

  20. Structure and Dynamics of the Liver Receptor Homolog 1-PGC1α Complex.

    PubMed

    Mays, Suzanne G; Okafor, C Denise; Tuntland, Micheal L; Whitby, Richard J; Dharmarajan, Venkatasubramanian; Stec, Józef; Griffin, Patrick R; Ortlund, Eric A

    2017-07-01

    Peroxisome proliferator-activated gamma coactivator 1- α (PGC1 α ) regulates energy metabolism by directly interacting with transcription factors to modulate gene expression. Among the PGC1 α binding partners is liver receptor homolog 1 (LRH-1; NR5A2), an orphan nuclear hormone receptor that controls lipid and glucose homeostasis. Although PGC1 α is known to bind and activate LRH-1, mechanisms through which PGC1 α changes LRH-1 conformation to drive transcription are unknown. Here, we used biochemical and structural methods to interrogate the LRH-1-PGC1 α complex. Purified, full-length LRH-1, as well as isolated ligand binding domain, bound to PGC1 α with higher affinity than to the coactivator, nuclear receptor coactivator-2 (Tif2), in coregulator peptide recruitment assays. We present the first crystal structure of the LRH-1-PGC1 α complex, which depicts several hydrophobic contacts and a strong charge clamp at the interface between these partners. In molecular dynamics simulations, PGC1 α induced correlated atomic motion throughout the entire LRH-1 activation function surface, which was dependent on charge-clamp formation. In contrast, Tif2 induced weaker signaling at the activation function surface than PGC1 α but promoted allosteric signaling from the helix 6/ β -sheet region of LRH-1 to the activation function surface. These studies are the first to probe mechanisms underlying the LRH-1-PGC1 α interaction and may illuminate strategies for selective therapeutic targeting of PGC1 α -dependent LRH-1 signaling pathways. Copyright © 2017 by The American Society for Pharmacology and Experimental Therapeutics.

  1. SiC-Based Miniature High-Temperature Cantilever Anemometer

    NASA Technical Reports Server (NTRS)

    Okojie, Robert S.; Fralick, Gustave; Saad, George J.

    2004-01-01

    The figure depicts a miniature cantilever-type anemometer that has been developed as a prototype of compact, relatively nonintrusive anemometers that can function at temperatures up to 600 C and that can be expected to be commercially mass-producible at low cost. The design of this anemometer, and especially the packaging aspect of the design, is intended to enable measurement of turbulence in the high-temperature, high-vibration environment of a turbine engine or in any similar environment. The main structural components of the anemometer include a single-crystal SiC cantilever and two polycrystalline SiC clamping plates, all made from chemical-vapor-deposited silicon carbide. Fabrication of these components from the same basic material eliminates thermal-expansion mismatch, which has introduced spurious thermomechanical stresses in cantilever-type anemometers of prior design. The clamping plates are heavily oxidized to improve electrical insulation at high temperature. A cavity that serves as a receptacle for the clamped end of the cantilever is etched into one end of one clamping plate. Trenches that collectively constitute a socket for a multipin electrical plug (for connection to external electronic circuitry) are etched into the opposite end of this clamping plate. Metal strips for electrical contact are deposited on one face of the other clamping plate. Piezoresistive single-crystal SiC thin-film strain gauges are etched in the n-type SiC epilayer in a Wheatstone-bridge configuration. Metal contact pads on the cantilever that extend into the clamping-receptacle area, are obtained by deposition and patterning using standard semiconductor photolithography and etching methods. The cantilever and the two clamping plates are assembled into a sandwich structure that is then clamped in a stainless-steel housing. The Wheatstone- bridge carrying SiC cantilever with the metal contact pads on the piezoresistors is slid into the receptacle in the bottom clamping plate. The top clamping plate is brought into contact with the bottom plate so that the narrow section of the metal strips on the top clamp plate aligns with the metal contact pads on the cantilever. When the parts are clamped together, the metal strips provide electrical connections between the Wheatstone-bridge contact points and the sides the trenches that constitute the socket for the multipin electrical plug. Hence, to connect the Wheatstone bridge to external circuitry for processing of the anemometer readout, one need only insert the plug in the socket. In operation, the cantilever end of the stainless-steel housing is mounted flush with an engine wall and the unclamped portion of the cantilever is exposed into the flow. The cantilever is deflected in direct proportion to the force induced by component of flow parallel to the engine wall and perpendicular to the broad exposed face of the cantilever. The maximum strain on the cantilever occurs at the clamped edge and is measured by the piezoresistors, which are located there. The corresponding changes in resistance manifest themselves in the output of the Wheatstone bridge.

  2. Mechanics of wafer bonding: Effect of clamping

    NASA Astrophysics Data System (ADS)

    Turner, K. T.; Thouless, M. D.; Spearing, S. M.

    2004-01-01

    A mechanics-based model is developed to examine the effects of clamping during wafer bonding processes. The model provides closed-form expressions that relate the initial geometry and elastic properties of the wafers to the final shape of the bonded pair and the strain energy release rate at the interface for two different clamping configurations. The results demonstrate that the curvature of bonded pairs may be controlled through the use of specific clamping arrangements during the bonding process. Furthermore, it is demonstrated that the strain energy release rate depends on the clamping configuration and that using applied loads usually leads to an undesirable increase in the strain energy release rate. The results are discussed in detail and implications for process development and bonding tool design are highlighted.

  3. Anatomic renal artery branch microdissection to facilitate zero-ischemia partial nephrectomy.

    PubMed

    Ng, Casey K; Gill, Inderbir S; Patil, Mukul B; Hung, Andrew J; Berger, Andre K; de Castro Abreu, Andre Luis; Nakamoto, Masahiko; Eisenberg, Manuel S; Ukimura, Osamu; Thangathurai, Duraiyah; Aron, Monish; Desai, Mihir M

    2012-01-01

    Robot-assisted and laparoscopic partial nephrectomies (PNs) for medial tumors are technically challenging even with the hilum clamped and, until now, were impossible to perform with the hilum unclamped. Evaluate whether targeted vascular microdissection (VMD) of renal artery branches allows zero-ischemia PN to be performed even for challenging medial tumors. A prospective cohort evaluation of 44 patients with renal masses who underwent robot-assisted or laparoscopic zero-ischemia PN either with anatomic VMD (group 1; n=22) or without anatomic VMD (group 2; n=22) performed by a single surgeon from April 2010 to January 2011. Zero-ischemia PN with VMD incorporates four maneuvers: (1) preoperative computed tomographic reconstruction of renal arterial branch anatomy, (2) anatomic dissection of targeted, tumor-specific tertiary or higher-order renal arterial branches, (3) neurosurgical aneurysm microsurgical bulldog clamp(s) for superselective tumor devascularization, and (4) transient, controlled reduction of blood pressure, if necessary. Baseline, perioperative, and postoperative data were collected prospectively. Group 1 tumors were larger (4.3 vs 2.6 cm; p=0.011), were more often hilar (41% vs 9%; p=0.09), were medial (59% and 23%; p=0.017), were closer to the hilum (1.46 vs 3.26 cm; p=0.0002), and had a lower C index score (2.1 vs 3.9; p=0.004) and higher RENAL nephrometry scores (7.7 vs 6.2; p=0.013). Despite greater complexity, no group 1 tumor required hilar clamping, and perioperative outcomes were similar to those of group 2: operating room time (4.7 and 4.1h), median blood loss (200 and 100ml), surgical margins for cancer (all negative), major complications (0% and 9%), and minor complications (18% and 14%). The median serum creatinine level was similar 2 mo postoperatively (1.2 and 1.3mg/dl). The study was limited by the relatively small sample size. Anatomic targeted dissection and superselective control of tumor-specific renal arterial branches facilitate zero-ischemia PN. Even challenging medial and hilar tumors can be excised without hilar clamping. Global surgical renal ischemia has been eliminated for most patients undergoing PN at our institution. Copyright © 2011 European Association of Urology. Published by Elsevier B.V. All rights reserved.

  4. Training Materials and Data Requirements for Unit Conduct of Fire Trainer (U-COFT) Training Test Support Plan

    DTIC Science & Technology

    1980-08-01

    DESIGN GROUP I GROUP II GROUP III (XMl Tank) (Chrysler UCOFT) (GE UCOFT) Pretest ...training plan design are that: 1. Tank Commanders and Gunners will have been transition trained on the "MI prior to assign- mint to a test group . Cuirrent...posttests on the XMI for each group . Also, Loaders and Drivers are required for the Group I device pretest , gunnery portion of the

  5. Automated Gun Laying System for Self-Propelled Artillery Weapons.

    DTIC Science & Technology

    1980-05-30

    model designed specifically to the requirements of a test bed system. The system configuration and characteristics were specified through a series of...proposed by the contractor was further defined, utilizing the M109 component information provided by the COTR. Math models were developed to predict system...data. The model used for the TB-I program did not have the capability for a remote reset function, hence it was necessary to instruct the crew (loader

  6. The CRRES Langmuir Probe and Fluxgate Magnetometer Instrument

    DTIC Science & Technology

    1989-05-09

    Gbntract tenager Branch Chief i r FOR THE COWhbDER RITA C. SAGALYN Division Director Qualified requestors may obtain additional copies from the... Computations 73 2.9 Sawtooth Generation 80 2.10 Bias Sweeps 84 2.11 Main Program Loader 96 2.12 Boom Deployment 98 2.13 General Utilities...separated by 100 meters. The other pair of conductors are cylindrical wire boom elements also in the spin plane that are separated by an effective

  7. Apple App Store as a Business Model Supporting U.S. Navy Requirements

    DTIC Science & Technology

    2011-10-25

    credit card identity information for charging the required $99/year fee. Actual payment of the fee is handled by putting the developer program...required for the App Store. Applications are submitted via iTunes Connect (http://itunesconnect.apple.com). Table 1 gives a list of the...still being processed by the iTunes Connect system.  Invalid Binary—Appears when a binary is received through the Application Loader and has been

  8. Radiation Dose Assessment for Military Personnel of the Enewetak Atoll Cleanup Project (1977-1980)

    DTIC Science & Technology

    2018-04-13

    population are detailed. 2.1 Enewetak Atoll Setting Enewetak Atoll (Figure 2) is approximately 23 by 17 statute miles with the long axis running ...Survey and stake the boundaries of soil excision areas; • Remove excess brush; • Excise (scrape surface with bulldozer blade ) the area and...JTGJ-2 and FRST members Survey instruments, soil sampling tools, dump trucks, bucket and backhoe loaders, water tank trucks, scrape blades

  9. 2012 ENFORCE. Project Manager Force Projection Briefing to ENFORCE

    DTIC Science & Technology

    2012-04-16

    M-ATV Logistics MRAP Vehicle Systems (Joint Program) •Navistar MaxxPro •GDLS RG-31 •BAE-TVS Caiman •BAE RG-33 SOCOM •BAE RG-33 SOCOM AUV...Handling Equipment • Container Handling Equipment •Loaders • Cranes •Dozers •Excavators Force Sustainment Systems •Cargo Airdrop Systems...Boruff DPM: Mr. Dennis Mazurek Mine Resistant Ambush Protected Vehicles PM: Mr. Carl Owens DPM: COL Jeffrey Carr Tactical

  10. INTERIOR VIEW, NORTH QUARRY, LOOKING WEST. IN THE FOREGROUND ON ...

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

    INTERIOR VIEW, NORTH QUARRY, LOOKING WEST. IN THE FOREGROUND ON THE FIRST BENCH, POWDER HILLS ARE PRIMED FOR DOLOMITE EXTRACTION. ON THE SECOND BENCH, THE DRILL TEAM IS LAYING OUT THE NEXT SHOTS. ON THE TOP BENCH, A 245 CATERPILLAR LOADER FILLS A 55-TON CATERPILLAR ROCK TRUCK WITH EXTRACTED DOLOMITE FOR TRANSPORT TO THE DOLOMITE CRUSHING AND SCREENING PLANT. - Wade Sand & Gravel Company, North Quarry, State Highway 78, Thomas, Jefferson County, AL

  11. Explicit frequency equations of free vibration of a nonlocal Timoshenko beam with surface effects

    NASA Astrophysics Data System (ADS)

    Zhao, Hai-Sheng; Zhang, Yao; Lie, Seng-Tjhen

    2018-02-01

    Considerations of nonlocal elasticity and surface effects in micro- and nanoscale beams are both important for the accurate prediction of natural frequency. In this study, the governing equation of a nonlocal Timoshenko beam with surface effects is established by taking into account three types of boundary conditions: hinged-hinged, clamped-clamped and clamped-hinged ends. For a hinged-hinged beam, an exact and explicit natural frequency equation is obtained. However, for clamped-clamped and clamped-hinged beams, the solutions of corresponding frequency equations must be determined numerically due to their transcendental nature. Hence, the Fredholm integral equation approach coupled with a curve fitting method is employed to derive the approximate fundamental frequency equations, which can predict the frequency values with high accuracy. In short, explicit frequency equations of the Timoshenko beam for three types of boundary conditions are proposed to exhibit directly the dependence of the natural frequency on the nonlocal elasticity, surface elasticity, residual surface stress, shear deformation and rotatory inertia, avoiding the complicated numerical computation.

  12. Non-invasive method and apparatus for measuring pressure within a pliable vessel

    NASA Technical Reports Server (NTRS)

    Shimizu, M. (Inventor)

    1983-01-01

    A non-invasive method and apparatus is disclosed for measuring pressure within a pliable vessel such as a blood vessel. The blood vessel is clamped by means of a clamping structure having a first portion housing a pressure sensor and a second portion extending over the remote side of the blood vessel for pressing the blood vessel into engagement with the pressure sensing device. The pressure sensing device includes a flat deflectable diaphragm portion arranged to engage a portion of the blood vessel flattened against the diaphragm by means of the clamp structure. In one embodiment, the clamp structure includes first and second semicylindrical members held together by retaining rings. In a second embodiment the clamp structure is of one piece construction having a solid semicylindrical portion and a hollow semicylindrical portion with a longitudinal slot in the follow semicylindrical portion through which a slip the blood vessel. In a third embodiment, an elastic strap is employed for clamping the blood vessel against the pressure sensing device.

  13. Term babies with delayed cord clamping: an approach in preventing anemia (.).

    PubMed

    Ertekin, Arif Aktug; Nihan Ozdemir, Nilufer; Sahinoglu, Zeki; Gursoy, Tugba; Erbil, Nazan; Kaya, Erdal

    2016-09-01

    We investigated the effects of delayed and early clamping of the cord on the hematologic status of the baby at birth and at the end of second month. Umbilical cord of 74 babies were clamped in the first 30 s (Group 1) and 76 were clamped at 90-120 s (Group 2). Levels of hemoglobin, hematocrit, iron and ferritin were analyzed from the umbilical cord blood at birth and from the venous samples at the end of second month. Hemoglobin, hematocrit, iron and ferritin levels of cord blood were similar in both groups. However, their levels other than ferritin were higher in Group 2 at the end of second month. Two babies had respiratory distress and twelve neonates received phototherapy in Group 2 whereas only five neonates received phototherapy in Group 1. Term babies to whom delayed cord clamping was performed had improved hematological parameters at the end of second month. Therefore, delaying cord clamping in these babies may be a favorible approach in preventing anemia.

  14. Vibration mode analysis of the proton exchange membrane fuel cell stack

    NASA Astrophysics Data System (ADS)

    Liu, B.; Liu, L. F.; Wei, M. Y.; Wu, C. W.

    2016-11-01

    Proton exchange membrane fuel cell (PEMFC) stacks usually undergo vibration during packing, transportation, and serving time, in particular for those used in the automobiles or portable equipment. To study the stack vibration response, based on finite element method (FEM), a mode analysis is carried out in the present paper. Using this method, we can distinguish the local vibration from the stack global modes, predict the vibration responses, such as deformed shape and direction, and discuss the effects of the clamping configuration and the clamping force magnitude on vibration modes. It is found that when the total clamping force remains the same, increasing the bolt number can strengthen the stack resistance to vibration in the clamping direction, but cannot obviously strengthen stack resistance to vibration in the translations perpendicular to clamping direction and the three axis rotations. Increasing the total clamping force can increase both of the stack global mode and the bolt local mode frequencies, but will decrease the gasket local mode frequency.

  15. Prosthetic helping hand

    NASA Technical Reports Server (NTRS)

    Vest, Thomas W. (Inventor); Carden, James R. (Inventor); Norton, William E. (Inventor); Belcher, Jewell G. (Inventor)

    1992-01-01

    A prosthetic device for below-the-elbow amputees, having a C-shaped clamping mechanism for grasping cylindrical objects, is described. The clamping mechanism is pivotally mounted to a cuff that fits on the amputee's lower arm. The present invention is utilized by placing an arm that has been amputated below the elbow into the cuff. The clamping mechanism then serves as a hand whenever it becomes necessary for the amputee to grasp a cylindrical object such as a handle, a bar, a rod, etc. To grasp the cylindrical object, the object is jammed against the opening in the C-shaped spring, causing the spring to open, the object to pass to the center of the spring, and the spring to snap shut behind the object. Various sizes of clamping mechanisms can be provided and easily interchanged to accommodate a variety of diameters. With the extension that pivots and rotates, the clamping mechanism can be used in a variety of orientations. Thus, this invention provides the amputee with a clamping mechanism that can be used to perform a number of tasks.

  16. Real-Time Kinetic Modeling of Voltage-Gated Ion Channels Using Dynamic Clamp

    PubMed Central

    Milescu, Lorin S.; Yamanishi, Tadashi; Ptak, Krzysztof; Mogri, Murtaza Z.; Smith, Jeffrey C.

    2008-01-01

    We propose what to our knowledge is a new technique for modeling the kinetics of voltage-gated ion channels in a functional context, in neurons or other excitable cells. The principle is to pharmacologically block the studied channel type, and to functionally replace it with dynamic clamp, on the basis of a computational model. Then, the parameters of the model are modified in real time (manually or automatically), with the objective of matching the dynamical behavior of the cell (e.g., action potential shape and spiking frequency), but also the transient and steady-state properties of the model (e.g., those derived from voltage-clamp recordings). Through this approach, one may find a model and parameter values that explain both the observed cellular dynamics and the biophysical properties of the channel. We extensively tested the method, focusing on Nav models. Complex Markov models (10–12 states or more) could be accurately integrated in real time at >50 kHz using the transition probability matrix, but not the explicit Euler method. The practicality of the technique was tested with experiments in raphe pacemaker neurons. Through automated real-time fitting, a Hodgkin-Huxley model could be found that reproduced well the action potential shape and the spiking frequency. Adding a virtual axonal compartment with a high density of Nav channels further improved the action potential shape. The computational procedure was implemented in the free QuB software, running under Microsoft Windows and featuring a friendly graphical user interface. PMID:18375511

  17. Tensil Film Clamps And Mounting Block For Viscoelastometers

    NASA Technical Reports Server (NTRS)

    Stoakley, Diane M.; St. Clair, Anne K.; Little, Bruce D.

    1989-01-01

    Set of clamps and mounting block developed for use in determining tensile moduli and damping properties of films in manually operated or automated commercial viscoelastometer. These clamps and block provide uniformity of sample gripping and alignment in instrument. Dependence on operator and variability of data greatly reduced.

  18. The 9-1-1 DNA Clamp Is Required for Immunoglobulin Gene Conversion▿

    PubMed Central

    Saberi, Alihossein; Nakahara, Makoto; Sale, Julian E.; Kikuchi, Koji; Arakawa, Hiroshi; Buerstedde, Jean-Marie; Yamamoto, Kenichi; Takeda, Shunichi; Sonoda, Eiichiro

    2008-01-01

    Chicken DT40 cells deficient in the 9-1-1 checkpoint clamp exhibit hypersensitivity to a variety of DNA-damaging agents. Although recent work suggests that, in addition to its role in checkpoint activation, this complex may play a role in homologous recombination and translesion synthesis, the cause of this hypersensitivity has not been studied thoroughly. The immunoglobulin locus of DT40 cells allows monitoring of homologous recombination and translesion synthesis initiated by activation-induced deaminase (AID)-dependent abasic sites. We show that both the RAD9−/− and RAD17−/− mutants exhibit substantially reduced immunoglobulin gene conversion. However, the level of nontemplated immunoglobulin point mutation increased in these mutants, a finding that is reminiscent of the phenotype resulting from the loss of RAD51 paralogs or Brca2. This suggests that the 9-1-1 complex does not play a central role in translesion synthesis in this context. Despite reduced immunoglobulin gene conversion, the RAD9−/− and RAD17−/− cells do not exhibit a prominent defect in double-strand break-induced gene conversion or a sensitivity to camptothecin. This suggests that the roles of Rad9 and Rad17 may be confined to a subset of homologous recombination reactions initiated by replication-stalling lesions rather than those associated with double-strand break repair. PMID:18662998

  19. 30 CFR 77.603 - Clamping of trailing cables to equipment.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Clamping of trailing cables to equipment. 77.603 Section 77.603 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR COAL... UNDERGROUND COAL MINES Trailing Cables § 77.603 Clamping of trailing cables to equipment. Trailing cables...

  20. 46 CFR 28.335 - Fuel systems.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... (2) Nonmetallic flexible hose is acceptable but must— (i) Not be used in lengths of more than 30... reinforced with wire braid. (iv) Be fitted with suitable, corrosion resistant, compression fittings; and (v) Be installed with two clamps at each end of the hose, if designed for use with clamps. Clamps must...

  1. 46 CFR 28.335 - Fuel systems.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... (2) Nonmetallic flexible hose is acceptable but must— (i) Not be used in lengths of more than 30... reinforced with wire braid. (iv) Be fitted with suitable, corrosion resistant, compression fittings; and (v) Be installed with two clamps at each end of the hose, if designed for use with clamps. Clamps must...

  2. 46 CFR 28.335 - Fuel systems.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... (2) Nonmetallic flexible hose is acceptable but must— (i) Not be used in lengths of more than 30... reinforced with wire braid. (iv) Be fitted with suitable, corrosion resistant, compression fittings; and (v) Be installed with two clamps at each end of the hose, if designed for use with clamps. Clamps must...

  3. 46 CFR 28.335 - Fuel systems.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... (2) Nonmetallic flexible hose is acceptable but must— (i) Not be used in lengths of more than 30... reinforced with wire braid. (iv) Be fitted with suitable, corrosion resistant, compression fittings; and (v) Be installed with two clamps at each end of the hose, if designed for use with clamps. Clamps must...

  4. An ideal clamping analysis for a cross-ply laminate

    NASA Technical Reports Server (NTRS)

    Valisetty, R. R.; Murthy, P. L. N.; Rehfield, L. W.

    1988-01-01

    Different elementary clamping models are discussed for a three layer crossply laminate to study the sensitivity of clamping to the definition of cross-sectional rotation. All of these models leave a considerable residual warping at the edges. Using a complimentary energy principle and principle of superposition, an analysis is conducted to reduce this residual warping. This led to the identification of exact interior solution corresponding to the ideal clamping. This study also suggests a presence of stress singularities at the corners and between different layers near the fixed edge.

  5. Clamp force and alignment checking device

    DOEpatents

    Spicer, John Patrick; Cai, Wayne W.; Chakraborty, Debejyo; Mink, Keith

    2017-04-11

    A check fixture measures a total clamp force applied by a welder device. The welder device includes a welding horn having a plurality of weld pads and welding anvil having a plurality of weld pads. The check fixture includes a base member operatively supporting a plurality of force sensors. The base member and the force sensors are received between the weld pads of the welding horn and the anvil pads of the welding anvil. Each force sensor is configured to measure an individual clamp force applied thereto by corresponding weld and anvil pads when the base member is received between the welding horn and the welding anvil and the welder device is in the clamped position. The individual clamp forces are used to determine whether the weld and/or anvil pads are worn or misaligned.

  6. Acoustic plane waves incident on an oblique clamped panel in a rectangular duct

    NASA Technical Reports Server (NTRS)

    Unz, H.; Roskam, J.

    1980-01-01

    The theory of acoustic plane waves incident on an oblique clamped panel in a rectangular duct was developed from basic theoretical concepts. The coupling theory between the elastic vibrations of the panel (plate) and the oblique incident acoustic plane wave in infinite space was considered in detail, and was used for the oblique clamped panel in the rectangular duct. The partial differential equation which governs the vibrations of the clamped panel (plate) was modified by adding to it stiffness (spring) forces and damping forces. The Transmission Loss coefficient and the Noise Reduction coefficient for oblique incidence were defined and derived in detail. The resonance frequencies excited by the free vibrations of the oblique finite clamped panel (plate) were derived and calculated in detail for the present case.

  7. Analytic description of the frictionally engaged in-plane bending process incremental swivel bending (ISB)

    NASA Astrophysics Data System (ADS)

    Frohn, Peter; Engel, Bernd; Groth, Sebastian

    2018-05-01

    Kinematic forming processes shape geometries by the process parameters to achieve a more universal process utilizations regarding geometric configurations. The kinematic forming process Incremental Swivel Bending (ISB) bends sheet metal strips or profiles in plane. The sequence for bending an arc increment is composed of the steps clamping, bending, force release and feed. The bending moment is frictionally engaged by two clamping units in a laterally adjustable bending pivot. A minimum clamping force hindering the material from slipping through the clamping units is a crucial criterion to achieve a well-defined incremental arc. Therefore, an analytic description of a singular bent increment is developed in this paper. The bending moment is calculated by the uniaxial stress distribution over the profiles' width depending on the bending pivot's position. By a Coulomb' based friction model, necessary clamping force is described in dependence of friction, offset, dimensions of the clamping tools and strip thickness as well as material parameters. Boundaries for the uniaxial stress calculation are given in dependence of friction, tools' dimensions and strip thickness. The results indicate that changing the bending pivot to an eccentric position significantly affects the process' bending moment and, hence, clamping force, which is given in dependence of yield stress and hardening exponent. FE simulations validate the model with satisfactory accordance.

  8. A novel monolithic piezoelectric actuated flexure-mechanism based wire clamp for microelectronic device packaging.

    PubMed

    Liang, Cunman; Wang, Fujun; Tian, Yanling; Zhao, Xingyu; Zhang, Hongjie; Cui, Liangyu; Zhang, Dawei; Ferreira, Placid

    2015-04-01

    A novel monolithic piezoelectric actuated wire clamp is presented in this paper to achieve fast, accurate, and robust microelectronic device packaging. The wire clamp has compact, flexure-based mechanical structure and light weight. To obtain large and robust jaw displacements and ensure parallel jaw grasping, a two-stage amplification composed of a homothetic bridge type mechanism and a parallelogram leverage mechanism was designed. Pseudo-rigid-body model and Lagrange approaches were employed to conduct the kinematic, static, and dynamic modeling of the wire clamp and optimization design was carried out. The displacement amplification ratio, maximum allowable stress, and natural frequency were calculated. Finite element analysis (FEA) was conducted to evaluate the characteristics of the wire clamp and wire electro discharge machining technique was utilized to fabricate the monolithic structure. Experimental tests were carried out to investigate the performance and the experimental results match well with the theoretical calculation and FEA. The amplification ratio of the clamp is 20.96 and the working mode frequency is 895 Hz. Step response test shows that the wire clamp has fast response and high accuracy and the motion resolution is 0.2 μm. High speed precision grasping operations of gold and copper wires were realized using the wire clamper.

  9. Double-stator electromagnetic pump having alignment ring and spine assembly

    DOEpatents

    Fanning, Alan Wayne; Olich, Eugene Ellsworth; Dahl, Leslie Roy; Patel, Mahadeo Ratilal

    1997-01-01

    A support structure for clamping the inner coils and inner lamination rings of an inner stator column of an electromagnetic induction pump to prevent damaging vibration. A spine assembly, including a base plate, a center post and a plurality of ribs, serves as the structural frame for the inner stator. Stacked alignment rings provide structure to the lamination rings and locate them concentrically around the spine assembly central axis. The alignment rings are made of a material having a high thermal expansion coefficient to compensate for the lower expansion of the lamination rings and, overall, provide an approximate match to the expansion of the inner flow duct. The net result is that the radial clamping provided by the duct around the stator iron is maintained (approximately) over a range of temperatures and operating conditions. Axial clamping of the inner stator structure is achieved via tie rods which run through grooves in the ribs and engage the base plate at the bottom of the inner stator and engage a clamping plate at the top. Slender tie rods and a flexible clamping plate are used to provide compliance in the axial clamping system to accommodate differential thermal growth (axially) between the tie rods and lamination ring elements without losing clamping force.

  10. Double-stator electromagnetic pump having alignment ring and spine assembly

    DOEpatents

    Fanning, A.W.; Olich, E.E.; Dahl, L.R.; Patel, M.R.

    1997-06-24

    A support structure for clamping the inner coils and inner lamination rings of an inner stator column of an electromagnetic induction pump to prevent damaging vibration is disclosed. A spine assembly, including a base plate, a center post and a plurality of ribs, serves as the structural frame for the inner stator. Stacked alignment rings provide structure to the lamination rings and locate them concentrically around the spine assembly central axis. The alignment rings are made of a material having a high thermal expansion coefficient to compensate for the lower expansion of the lamination rings and, overall, provide an approximate match to the expansion of the inner flow duct. The net result is that the radial clamping provided by the duct around the stator iron is maintained (approximately) over a range of temperatures and operating conditions. Axial clamping of the inner stator structure is achieved via tie rods which run through grooves in the ribs and engage the base plate at the bottom of the inner stator and engage a clamping plate at the top. Slender tie rods and a flexible clamping plate are used to provide compliance in the axial clamping system to accommodate differential thermal growth (axially) between the tie rods and lamination ring elements without losing clamping force. 12 figs.

  11. [Analysis of brain hemometabolism behavior during carotid endarterectomy with temporary clamping.].

    PubMed

    Duval Neto, Gastão Fernandes; Niencheski, Augusto H

    2004-04-01

    Carotid endarterectomy with temporary clamping changes cerebral blood flow and cerebral metabolic oxygen demand ratio with consequent oligemic hypoxia or hemometabolic uncoupling. This study aimed at identifying changes in brain hemometabolism, evaluated through changes in oxyhemoglobin saturation in internal jugular vein bulb (SvjO2) during carotid endarterectomy with clamping, and at correlating these changes with potentially interfering factors, mainly end tidal CO2 pressure (P ET CO2) and cerebral perfusion pressure (CPP). Sixteen patients with unilateral carotid stenotic disease scheduled to carotid endarterectomy with carotid arterial clamping were enrolled in this study. Parameters including internal jugular bulb oxyhemoglobin saturation, stump pressure and end tidal CO2 pressure were measured at the following moments: M1 - pre-clamping; M2 - 3 minutes after clamping; M3 - pre-unclamping; M4 - post-unclamping). The comparison among SvjO2 (%, mean +/- SD) in all studied periods has shown differences between those recorded in moments M1 (52.25 +/- 7.87) and M2 (47.43 +/- 9.19). This initial decrease stabilized during temporary clamping, showing decrease in the comparison between M2 and M3 (46.56 +/- 9.25), without statistical significance (p = ns). At post-unclamping, M4 (47.68 +/- 9.12), SvjO2 was increased as compared to M2 and M3 clamping stages, however it was still lower than that of pre-clamping stage M1.(M4 x M1 - p < 0.04) This SvjO2 decrease was followed by significant cerebral perfusion pressure (stump pressure) decrease. Factors influencing this brain hemometabolic uncoupling trend were correlated to P ET CO2. The comparison between CPP and SvjO2 showed weak correlation devoid of statistical significance. In the conditions of our study, SvjO2 measurement is a fast and effective way of clinically monitoring changes in CBF/CMRO2 ratio. Temporary carotid clamping implies in a trend towards brain hemometabolic uncoupling and, as a consequence, to oligemic ischemia; cerebral perfusion pressure does not assesses brain hemometabolic status (CBF and CMRO2 ratio); hypocapnia, may lead to brain hemometabolic uncoupling; P ET CO2 monitoring is an innocuous and efficient way to indirectly monitor PaCO2 preventing inadvertent hypocapnia and its deleterious effects on CBF/CMRO2 ratio during temporary carotid clamping.

  12. GIDL: a rule based expert system for GenBank Intelligent Data Loading into the Molecular Biodiversity database

    PubMed Central

    2012-01-01

    Background In the scientific biodiversity community, it is increasingly perceived the need to build a bridge between molecular and traditional biodiversity studies. We believe that the information technology could have a preeminent role in integrating the information generated by these studies with the large amount of molecular data we can find in bioinformatics public databases. This work is primarily aimed at building a bioinformatic infrastructure for the integration of public and private biodiversity data through the development of GIDL, an Intelligent Data Loader coupled with the Molecular Biodiversity Database. The system presented here organizes in an ontological way and locally stores the sequence and annotation data contained in the GenBank primary database. Methods The GIDL architecture consists of a relational database and of an intelligent data loader software. The relational database schema is designed to manage biodiversity information (Molecular Biodiversity Database) and it is organized in four areas: MolecularData, Experiment, Collection and Taxonomy. The MolecularData area is inspired to an established standard in Generic Model Organism Databases, the Chado relational schema. The peculiarity of Chado, and also its strength, is the adoption of an ontological schema which makes use of the Sequence Ontology. The Intelligent Data Loader (IDL) component of GIDL is an Extract, Transform and Load software able to parse data, to discover hidden information in the GenBank entries and to populate the Molecular Biodiversity Database. The IDL is composed by three main modules: the Parser, able to parse GenBank flat files; the Reasoner, which automatically builds CLIPS facts mapping the biological knowledge expressed by the Sequence Ontology; the DBFiller, which translates the CLIPS facts into ordered SQL statements used to populate the database. In GIDL Semantic Web technologies have been adopted due to their advantages in data representation, integration and processing. Results and conclusions Entries coming from Virus (814,122), Plant (1,365,360) and Invertebrate (959,065) divisions of GenBank rel.180 have been loaded in the Molecular Biodiversity Database by GIDL. Our system, combining the Sequence Ontology and the Chado schema, allows a more powerful query expressiveness compared with the most commonly used sequence retrieval systems like Entrez or SRS. PMID:22536971

  13. Spectral infrared hemispherical reflectance measurements for LDEF tray clamps

    NASA Technical Reports Server (NTRS)

    Wood, Bobby E.; Cromwell, Brian K.; Pender, Charles W.; Shepherd, Seth D.

    1992-01-01

    This paper describes infrared hemispherical reflectance measurements (2-15 microns) that were made on 58 chromic acid anodized tray clamps retrieved from the LDEF spacecraft. These clamps were used for maintaining the experiments in place and were located at various locations about the spacecraft. Changes in reflectance of the tray clamps at these locations were compared with atomic oxygen fluxes at the same locations. A decrease in absorption band depth was seen for the surfaces exposed to space indicating that there was some surface layer erosion. In all of the surfaces measured, little evidence of contamination was observed and none of the samples showed evidence of the brown nicotine stain that was so prominent in other experiments. Total emissivity values were calculated for both exposed and unexposed tray clamp surfaces. Only small differences, usually less than 1 percent, were observed. The spectral reflectances were measured using a hemi-ellipsoidal mirror reflectometer matched with an interferometer spectrometer. The rapid scanning capability of the interferometer allowed the reflectance measurements to be made in a timely fashion. The ellipsoidal mirror has its two foci separated by 2 inches and located on the major axis. A blackbody source was located at one focus while the tray clamp samples were located at the conjugate focus. The blackbody radiation was modulated and then focused by the ellipsoid onto the tray clamps. Radiation reflected from the tray clamp was sampled by the interferometer by viewing through a hole in the ellipsoid. A gold mirror (reflectance approximately 98 percent) was used as the reference surface.

  14. Cerebral hemodynamic changes and electroencephalography during carotid endarterectomy

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

    Algotsson, L.; Messeter, K.; Rehncrona, S.

    Some patients undergoing endarterectomy for occlusive carotid artery disease run a risk of brain ischemia during cross-clamping of the artery. The present study of 15 patients was undertaken to evaluate changes in cerebral blood flow (CBF), as measured with an intravenous (IV) tracer (133Xenon) technique, and to relate CBF changes to changes in the electroencephalogram (EEG). CBF was measured before and after induction of anesthesia, during cross-clamping of the carotid artery, after release of the clamps, and at 24 hours after the operation. All the patients were anesthetized with methohexitone, fentanyl, and nitrous oxide and oxygen. EEG was continuously recordedmore » during the operation. Carotid artery shunts were not used. In 8 patients, cross-clamping of the carotid artery did not influence the EEG. In this group of patients, induction of anesthesia caused a 38% decrease in CBF, which presumably reflects the normal reaction to the anesthetic agent given. There were no further changes in CBF during cross-clamping. In 7 patients, the EEG showed signs of deterioration during the intraoperative vascular occlusion. In these patients, anesthesia did not cause any CBF change, whereas cross-clamping the artery induced a 33% decrease in CBF. In individual patients, the severity of EEG changes correlated with the decrease in CBF. The absence of a change in CBF by anesthesia and a decrease due to cross-clamping of the carotid artery may be explained by the presence of a more advanced cerebrovascular disease and an insufficiency to maintain CBF during cross-clamping.« less

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

    Jia, Xiaofei; Singh, Rajendra; Homann, Stefanie

    The HIV-1 protein Nef inhibits antigen presentation by class I major histocompatibility complex (MHC-I). We determined the mechanism of this activity by solving the crystal structure of a protein complex comprising Nef, the MHC-I cytoplasmic domain (MHC-I CD) and the {mu}1 subunit of the clathrin adaptor protein complex 1. A ternary, cooperative interaction clamps the MHC-I CD into a narrow binding groove at the Nef-{mu}1 interface, which encompasses the cargo-recognition site of {mu}1 and the proline-rich strand of Nef. The Nef C terminus induces a previously unobserved conformational change in {mu}1, whereas the N terminus binds the Nef core tomore » position it optimally for complex formation. Positively charged patches on {mu}1 recognize acidic clusters in Nef and MHC-I. The structure shows how Nef functions as a clathrin-associated sorting protein to alter the specificity of host membrane trafficking and enable viral evasion of adaptive immunity.« less

  16. Design and Construction of a Small Whole Body Inhalation Chamber

    DTIC Science & Technology

    2010-07-30

    by 12 7/8 inch high) were solvent welded onto this base using Weldon 4 (Ridout Plastics, San Diego CA). On the inside of each side wall a 2 inch...together using one inch spring clamps (Just Clamps, model 616,Atlanta Ga) on their respective flanges. These clamps were spaced evenly around the

  17. Radiator debris removing apparatus and work machine using same

    DOEpatents

    Martin, Kevin L [Washburn, IL; Elliott, Dwight E [Chillicothe, IL

    2008-09-02

    A radiator assembly includes a finned radiator core and a debris removing apparatus having a compressed air inlet and at least one compressed air outlet configured to direct compressed air through the radiator core. A work machine such as a wheel loader includes a radiator and a debris removing apparatus coupled with on-board compressed air and having at least one pressurized gas outlet configured to direct a gas toward the face of the radiator.

  18. Economic analysis of a small-sized combined heat and power plant using forest biomass in the Republic of Korea

    Treesearch

    Yeongwan Seo; Han-Sup Han; Edward M. (Ted) Bilek; Jungkee Choi; Dusong Cha; Jungsoo Lee

    2017-01-01

    Economic analysis was conducted on the feasibility of operating a small-sized (500kW/hour) gasification power plant producing heat and electricity in a rural town surrounded by forests in the Republic of Korea. Cost factors that were considered over the plant’s 20-year life included wood procurement, a wood grab loader, a chipper, a chip dryer, a gasifier, a generator...

  19. Unit Reference Sheet (URS) Cost Methodology.

    DTIC Science & Technology

    1980-08-01

    LAUNCHER MONORAIL GUIDED MISSILE: W/E (NIKE-HERCULES) L45740 LAUNCHER TUBULAR GUIDED MISSILE: (TOW) L45757 LAUNCHER ZERO LENGTH GUIDED MISSILE: (IMP-HAWK...L76762 LOADER TRANSPORTER GUIDED MISSILE: W/E (HAWK) M57503 MOBILE TARGET TRACKING SYSTEM: USED TO SUPPORT MQM 34 ( FIRE BEE) M57549 MOBILITY KIT GUIDED...HIGH RATE THREE BARREL W/E J96479 GUN AUTOMATIC 20 MILLIMETER: GAS OPERATED MANUAL OR ELECT FIRED J96481 GUN AUTOMATIC 20 MILLIMETER: ELECTRIC J96694 GUN

  20. Analysis of impactor residues in tray clamps from the Long Duration Exposure Facility. Part 2: Clamps from Bay B of the satellite

    NASA Technical Reports Server (NTRS)

    Bernhard, Ronald P.; Zolensky, Michael E.

    1994-01-01

    The Long Duration Exposure Facility (LDEF) was placed in low-Earth orbit (LEO) in 1984 and recovered 5.7 years later. The LDEF was host to several individual experiments specifically designed to characterize critical aspects of meteoroid and debris environment in LEO. However, it was realized from the beginning that the most efficient use of the satellite would be to examine the entire surface for impact features. In this regard, particular interest centered on common exposed materials that faced in all LDEF pointing directions. Among the most important of these materials was the tray clamps. Therefore, in an effort to better understand the nature of particulates in LEO and their effects on spacecraft hardware, residues found in impact features on LDEF tray clamp surfaces are being analyzed. This catalog presents all data from clamps from Bay B of the LDEF. NASA Technical Memorandum 104759 has cataloged impacts that occurred on Bay B (published March 1993). Subsequent catalogs will include clamps from succeeding bays of the satellite.

  1. Micromolded PDMS planar electrode allows patch clamp electrical recordings from cells.

    PubMed

    Klemic, Kathryn G; Klemic, James F; Reed, Mark A; Sigworth, Fred J

    2002-06-01

    The patch clamp method measures membrane currents at very high resolution when a high-resistance 'gigaseal' is established between the glass microelectrode and the cell membrane (Pflugers Arch. 391 (1981) 85; Neuron 8 (1992) 605). Here we describe the first use of the silicone elastomer, poly(dimethylsiloxane) (PDMS), for patch clamp electrodes. PDMS is an attractive material for patch clamp recordings. It has low dielectric loss and can be micromolded (Annu. Rev. Mat. Sci. 28 (1998) 153) into a shape that mimics the tip of the glass micropipette. Also, the surface chemistry of PDMS may be altered to mimic the hydrophilic nature of glass (J. Appl. Polym. Sci. 14 (1970) 2499; Annu. Rev. Mat. Sci. 28 (1998) 153), thereby allowing a high-resistance seal to a cell membrane. We present a planar electrode geometry consisting of a PDMS partition with a small aperture sealed between electrode and bath chambers. We demonstrate that a planar PDMS patch electrode, after oxidation of the elastomeric surface, permits patch clamp recording on Xenopus oocytes. Our results indicate the potential for high-throughput patch clamp recording with a planar array of PDMS electrodes.

  2. Linear ultrasonic motor for absolute gravimeter.

    PubMed

    Jian, Yue; Yao, Zhiyuan; Silberschmidt, Vadim V

    2017-05-01

    Thanks to their compactness and suitability for vacuum applications, linear ultrasonic motors are considered as substitutes for classical electromagnetic motors as driving elements in absolute gravimeters. Still, their application is prevented by relatively low power output. To overcome this limitation and provide better stability, a V-type linear ultrasonic motor with a new clamping method is proposed for a gravimeter. In this paper, a mechanical model of stators with flexible clamping components is suggested, according to a design criterion for clamps of linear ultrasonic motors. After that, an effect of tangential and normal rigidity of the clamping components on mechanical output is studied. It is followed by discussion of a new clamping method with sufficient tangential rigidity and a capability to facilitate pre-load. Additionally, a prototype of the motor with the proposed clamping method was fabricated and the performance tests in vertical direction were implemented. Experimental results show that the suggested motor has structural stability and high dynamic performance, such as no-load speed of 1.4m/s and maximal thrust of 43N, meeting the requirements for absolute gravimeters. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Deuterium-Tritium Beta-Layering Within a National Ignition Facility Scale Polymer Target in the LANL Cryogenic Pressure Loader

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

    Ebey, Peter S.; Dole, James M.; Geller, Drew A.

    2005-11-15

    Beta-layering, the process of beta-decay heat-driven mass redistribution, has been demonstrated in a deuterium-tritium (D-T)-filled polymer sphere of the type required for fusion ignition experiments at the National Ignition Facility. This is the first report, to the best of the authors' knowledge, of a D-T layer formed in a permeation-filled sphere. The 2-mm-diam sphere was filled with D-T by permeation; cooled to cryogenic temperatures while in the high-pressure permeation vessel; and, while cold, removed to an optical axis where the D-T was frozen, melted, and beta-layered in a series of experiments over several weeks' time. This work was performed inmore » the Los Alamos National Laboratory cryogenic pressure loader system. The beta-layering time constant was 24.0 {+-} 2.5 min, less than the theoretical value of 26.8 min, and not showing the significant increase due to build-up of {sup 3}He often observed in beta-layered samples. Supercooling of the liquid D-T was observed. Neither the polymer target nor its tenting material showed visual signs of degradation after 5 weeks of exposure to D-T. Small external thermal gradients were used to shift the D-T material back and forth within the sphere.« less

  4. Slickenside formation by surface melting during the mechanical excavation of rock

    NASA Astrophysics Data System (ADS)

    Spray, John G.

    This work discusses the nature and origin of slickensides generated by the impingement of high-carbon steel teeth on sandstone during the mechanical excavation of boulders by back shovel and front loader. The slickensides show a number of morphological features that can be related to the direction and sense of tooth displacement, including striations, carrot-shaped grooves, curved fractures and steps. Scanning and transmission electron microscopy reveal that the slickensides comprise a layer of Fe-enriched, glass-bonded gouge (≤150 μm thick). The estimated shear stresses and velocities realized at the tooth-rock interface (175 MPa at 1 m s -1 for the back shovel and 100 MPa at 2 m s -1 for the front loader) indicate that a heat production of 150-200 MW m -2 and mean surface temperature of 1400-1700°C were achieved. XRF and microprobe analyses confirm that localized bulk melting and Fe-enrichment of the surface occurred during slip. The excavator-generated slickensides provide an analogue for the effects of a single co-seismic event in the evolution of a shallow, relatively dry fault surface. From a simple consideration of energetics under these conditions, it is apparent that localized surface melting should be commonplace. Such an analogue may be useful as a guide for recognizing melt features in natural slickensides, features that would otherwise tend to be obscured during lengthier periods of interseismic surface modification.

  5. Intermittent chest tube clamping may shorten chest tube drainage and postoperative hospital stay after lung cancer surgery: a propensity score matching analysis.

    PubMed

    Yan, Shi; Wang, Xing; Wang, Yaqi; Lv, Chao; Wang, Yuzhao; Wang, Jia; Yang, Yue; Wu, Nan

    2017-12-01

    Postoperative pleural drainage markedly influences the length of hospital stay and the financial costs of medical care. The safety of chest tube clamping before removal has been documented. This study aims to determine if intermittent chest tube clamping shortens the duration of chest tube drainage and hospital stay after lung cancer surgery. We retrospectively analyzed 285 consecutive patients with operable lung cancer treated using lobectomy and systematic mediastinal lymphadenectomy. The chest tube management protocol in our institution was changed in January 2014, and thus, 222 patients (clamping group) were managed with intermittent chest tube clamping, while 63 patients (control group) were managed with a traditional protocol. Propensity score matching at a 1:1 ratio was applied to balance variables potentially affecting the duration of chest tube drainage. Analyses were performed to compare drainage duration and postoperative hospital stay between the two groups in the matched cohort. Multivariate logistic regression analyses were performed to predict the factors associated with chest tube drainage duration. The rates of thoracocentesis after chest tube removal were similar between the clamping and control groups in the whole cohort (0.5% vs. 1.6%, P=0.386). The rates of pyrexia were also comparable in the two groups (2.3% vs. 3.2%, P=0.685). After propensity score matching, 61 cases remained in each group. Both chest tube drainage duration (3.9 vs. 4.8 days, P=0.001) and postoperative stay (5.7 vs. 6.4 days, P=0.025) were significantly shorter in the clamping group than in the control group. Factors significantly associated with shorter chest tube drainage duration were female sex, chest tube clamping, left lobectomy, and video-assisted thoracoscopic surgery (VATS) (P<0.05). Intermittent postoperative chest tube clamping may decrease the duration of chest tube drainage and postoperative hospital stay while maintaining patient safety.

  6. NIMBY, CLAMP, and the location of new nuclear-related facilities: U.S. national and 11 site-specific surveys.

    PubMed

    Greenberg, Michael R

    2009-09-01

    Public and political opposition have made finding locations for new nuclear power plants, waste management, and nuclear research and development facilities a challenge for the U.S. government and the nuclear industry. U.S. government-owned properties that already have nuclear-related activities and commercial nuclear power generating stations are logical locations. Several studies and utility applications to the Nuclear Regulatory Commission suggest that concentrating locations at major plants (CLAMP) has become an implicit siting policy. We surveyed 2,101 people who lived within 50 miles of 11 existing major nuclear sites and 600 who lived elsewhere in the United States. Thirty-four percent favored CLAMP for new nuclear power plants, 52% for waste management facilities, and 50% for new nuclear laboratories. College educated, relatively affluent male whites were the strongest CLAMP supporters. They disproportionately trusted those responsible for the facilities and were not worried about existing nuclear facilities or other local environmental issues. Notably, they were concerned about continuing coal use. Not surprisingly, CLAMP proponents tended to be familiar with their existing local nuclear site. In short, likely CLAMP sites have a large and politically powerful core group to support a CLAMP policy. The challenge to proponents of nuclear technologies will be to sustain this support and expand the base among those who clearly are less connected and receptive to new nearby sites.

  7. Piping Connector

    NASA Technical Reports Server (NTRS)

    1993-01-01

    A complex of high pressure piping at Stennis Space Center carries rocket propellants and other fluids/gases through the Center's Component Test Facility. Conventional clamped connectors tend to leak when propellant lines are chilled to extremely low temperatures. Reflange, Inc. customized an existing piping connector to include a secondary seal more tolerant of severe thermal gradients for Stennis. The T-Con connector solved the problem, and the company is now marketing a commercial version that permits testing, monitoring or collecting any emissions that may escape the primary seal during severe thermal transition.

  8. Tensile film clamps and mounting block for the rheovibron and autovibron viscoelastometer

    NASA Technical Reports Server (NTRS)

    Stoakley, Diane M. (Inventor); St.clair, Anne K. (Inventor); Little, Bruce D. (Inventor)

    1989-01-01

    A set of film clamps and a mounting block for use in the determination of tensile modulus and damping properties of films in a manually operated or automated Rheovibron is diagrammed. These clamps and mounting block provide uniformity of sample gripping and alignment in the instrument. Operator dependence and data variability are greatly reduced.

  9. Plastic Clamp Retains Clevis Pin

    NASA Technical Reports Server (NTRS)

    Cortes, R. G.

    1983-01-01

    Plastic clamp requires no special installation or removal tools. Clamp slips easily over end of pin. Once engaged in groove, holds pin securely. Installed and removed easily without special tools - screwdriver or putty knife adequate for prying out of groove. Used to retain bearings, rollers pulleys, other parts that rotate. Applications include slowly and intermittently rotating parts in appliances.

  10. Validation of simple indexes to assess insulin sensitivity during pregnancy in Wistar and Sprague-Dawley rats.

    PubMed

    Cacho, J; Sevillano, J; de Castro, J; Herrera, E; Ramos, M P

    2008-11-01

    Insulin resistance plays a role in the pathogenesis of diabetes, including gestational diabetes. The glucose clamp is considered the gold standard for determining in vivo insulin sensitivity, both in human and in animal models. However, the clamp is laborious, time consuming and, in animals, requires anesthesia and collection of multiple blood samples. In human studies, a number of simple indexes, derived from fasting glucose and insulin levels, have been obtained and validated against the glucose clamp. However, these indexes have not been validated in rats and their accuracy in predicting altered insulin sensitivity remains to be established. In the present study, we have evaluated whether indirect estimates based on fasting glucose and insulin levels are valid predictors of insulin sensitivity in nonpregnant and 20-day-pregnant Wistar and Sprague-Dawley rats. We have analyzed the homeostasis model assessment of insulin resistance (HOMA-IR), the quantitative insulin sensitivity check index (QUICKI), and the fasting glucose-to-insulin ratio (FGIR) by comparing them with the insulin sensitivity (SI(Clamp)) values obtained during the hyperinsulinemic-isoglycemic clamp. We have performed a calibration analysis to evaluate the ability of these indexes to accurately predict insulin sensitivity as determined by the reference glucose clamp. Finally, to assess the reliability of these indexes for the identification of animals with impaired insulin sensitivity, performance of the indexes was analyzed by receiver operating characteristic (ROC) curves in Wistar and Sprague-Dawley rats. We found that HOMA-IR, QUICKI, and FGIR correlated significantly with SI(Clamp), exhibited good sensitivity and specificity, accurately predicted SI(Clamp), and yielded lower insulin sensitivity in pregnant than in nonpregnant rats. Together, our data demonstrate that these indexes provide an easy and accurate measure of insulin sensitivity during pregnancy in the rat.

  11. Different methods of hilar clamping during partial nephrectomy: Impact on renal function.

    PubMed

    Lee, Jeong Woo; Kim, Hwanik; Choo, Minsoo; Park, Yong Hyun; Ku, Ja Hyeon; Kim, Hyeon Hoe; Kwak, Cheol

    2014-03-01

    To evaluate the impact of different hilar clamping methods on changes in renal function after partial nephrectomy. We analyzed the clinical data of 369 patients who underwent partial nephrectomy for a single renal tumor of size ≤4.0 cm and a normal contralateral kidney. Patients were separated into three groups depending on hilar clamping method: non-clamping, cold ischemia and warm ischemia. Estimated glomerular filtration rate was examined at preoperative, nadir and 1 year postoperatively. Percent change in estimated glomerular filtration rate was used as the parameter to assess the renal functional outcome. Percent change in nadir estimated glomerular filtration rate in the non-clamping group was significantly less compared with the cold ischemia and warm ischemia groups (P < 0.001). However, no significant differences among the groups were noted in percent change of estimated glomerular filtration rate at 1 year (P = 0.348). The cold ischemia group had a similar serial change of postoperative renal function compared with the warm ischemia group. Percent change in 1-year estimated glomerular filtration rate increased with increasing ischemia time in the cold ischemia (P for trend = 0.073) and warm ischemia groups (P for trend = 0.010). On multivariate analysis, hilar clamping (both warm ischemia and cold ischemia) were significantly associated with percent change in nadir estimated glomerular filtration rate, but not in 1-year estimated glomerular filtration rate. Non-clamping partial nephrectomy results in a lower percent change in nadir estimated glomerular filtration rate, whereas it carries an estimated glomerular filtration rate change at 1 year that is similar to partial nephrectomy with cold ischemia and warm ischemia. Cold ischemia and warm ischemia provide a similar effect on renal function. Therefore, when hilar clamping is required, minimization of ischemia time is necessary. © 2013 The Japanese Urological Association.

  12. A-Mating-Type Gene Expression Can Drive Clamp Formation in the Bipolar Mushroom Pholiota microspora (Pholiota nameko) ▿

    PubMed Central

    Yi, Ruirong; Mukaiyama, Hiroyuki; Tachikawa, Takashi; Shimomura, Norihiro; Aimi, Tadanori

    2010-01-01

    In the bipolar basidiomycete Pholiota microspora, a pair of homeodomain protein genes located at the A-mating-type locus regulates mating compatibility. In the present study, we used a DNA-mediated transformation system in P. microspora to investigate the homeodomain proteins that control the clamp formation. When a single homeodomain protein gene (A3-hox1 or A3-hox2) from the A3 monokaryon strain was transformed into the A4 monokaryon strain, the transformants produced many pseudoclamps but very few clamps. When two homeodomain protein genes (A3-hox1 and A3-hox2) were transformed either separately or together into the A4 monokaryon, the ratio of clamps to the clamplike cells in the transformants was significantly increased to ca. 50%. We therefore concluded that the gene dosage of homeodomain protein genes is important for clamp formation. When the sip promoter was connected to the coding region of A3-hox1 and A3-hox2 and the fused fragments were introduced into NGW19-6 (A4), the transformants achieved more than 85% clamp formation and exhibited two nuclei per cell, similar to the dikaryon (NGW12-163 × NGW19-6). The results of real-time reverse transcription-PCR confirmed that sip promoter activity is greater than that of the native promoter of homeodomain protein genes in P. microspora. Thus, we concluded that nearly 100% clamp formation requires high expression levels of homeodomain protein genes and that altered expression of the A-mating-type genes alone is sufficient to drive true clamp formation. PMID:20453073

  13. Safety evaluation of large external fixation clamps and frames in a magnetic resonance environment.

    PubMed

    Luechinger, Roger; Boesiger, Peter; Disegi, John A

    2007-07-01

    Large orthopedic external fixation clamps and related components were evaluated for force, torque, and heating response when subjected to the strong electromagnetic fields of magnetic-resonance (MR) imaging devices. Forces induced by a 3-Tesla (T) MR scanner were compiled for newly designed nonmagnetic clamps and older clamps that contained ferromagnetic components. Heating trials were performed in a 1.5 and in a 3 T MR scanner with two assembled external fixation frames. Forces of the newly designed clamps were more than a factor 2 lower as the gravitational force on the device whereas, magnetic forces on the older devices showed over 10 times the force induced by earth acceleration of gravity. No torque effects could be found for the newly designed clamps. Temperature measurements at the tips of Schanz screws in the 1.5 T MR scanner showed a rise of 0.7 degrees C for a pelvic frame and of 2.1 degrees C for a diamond knee bridge frame when normalized to a specific absorption rate (SAR) of 2 W/kg. The normalized temperature increases in the 3 T MR scanner were 0.9 degrees C for the pelvic frame and 1.1 degrees C for the knee bridge frame. Large external fixation frames assembled with the newly designed clamps (390 Series Clamps), carbon fiber reinforced rods, and implant quality 316L stainless steel Schanz screws met prevailing force and torque limits when tested in a 3-T field, and demonstrated temperature increase that met IEC-60601 guidelines for extremities. The influence of frame-induced eddy currents on the risk of peripheral nerve stimulation was not investigated. Copyright 2006 Wiley Periodicals, Inc.

  14. Substrate clamping effects on irreversible domain wall dynamics in lead zirconate titanate thin films.

    PubMed

    Griggio, F; Jesse, S; Kumar, A; Ovchinnikov, O; Kim, H; Jackson, T N; Damjanovic, D; Kalinin, S V; Trolier-McKinstry, S

    2012-04-13

    The role of long-range strain interactions on domain wall dynamics is explored through macroscopic and local measurements of nonlinear behavior in mechanically clamped and released polycrystalline lead zirconate-titanate (PZT) films. Released films show a dramatic change in the global dielectric nonlinearity and its frequency dependence as a function of mechanical clamping. Furthermore, we observe a transition from strong clustering of the nonlinear response for the clamped case to almost uniform nonlinearity for the released film. This behavior is ascribed to increased mobility of domain walls. These results suggest the dominant role of collective strain interactions mediated by the local and global mechanical boundary conditions on the domain wall dynamics. The work presented in this Letter demonstrates that measurements on clamped films may considerably underestimate the piezoelectric coefficients and coupling constants of released structures used in microelectromechanical systems, energy harvesting systems, and microrobots.

  15. Launch Lock Assemblies Including Axial Gap Amplification Devices and Spacecraft Isolation Systems Including the Same

    NASA Technical Reports Server (NTRS)

    Barber, Tim Daniel (Inventor); Hindle, Timothy (Inventor); Young, Ken (Inventor); Davis, Torey (Inventor)

    2014-01-01

    Embodiments of a launch lock assembly are provided, as are embodiments of a spacecraft isolation system including one or more launch lock assemblies. In one embodiment, the launch lock assembly includes first and second mount pieces, a releasable clamp device, and an axial gap amplification device. The releasable clamp device normally maintains the first and second mount pieces in clamped engagement; and, when actuated, releases the first and second mount pieces from clamped engagement to allow relative axial motion there between. The axial gap amplification device normally residing in a blocking position wherein the gap amplification device obstructs relative axial motion between the first and second mount pieces. The axial gap amplification device moves into a non-blocking position when the first and second mount pieces are released from clamped engagement to increase the range of axial motion between the first and second mount pieces.

  16. Structural basis of human PCNA sliding on DNA

    NASA Astrophysics Data System (ADS)

    de March, Matteo; Merino, Nekane; Barrera-Vilarmau, Susana; Crehuet, Ramon; Onesti, Silvia; Blanco, Francisco J.; de Biasio, Alfredo

    2017-01-01

    Sliding clamps encircle DNA and tether polymerases and other factors to the genomic template. However, the molecular mechanism of clamp sliding on DNA is unknown. Using crystallography, NMR and molecular dynamics simulations, here we show that the human clamp PCNA recognizes DNA through a double patch of basic residues within the ring channel, arranged in a right-hand spiral that matches the pitch of B-DNA. We propose that PCNA slides by tracking the DNA backbone via a `cogwheel' mechanism based on short-lived polar interactions, which keep the orientation of the clamp invariant relative to DNA. Mutation of residues at the PCNA-DNA interface has been shown to impair the initiation of DNA synthesis by polymerase δ (pol δ). Therefore, our findings suggest that a clamp correctly oriented on DNA is necessary for the assembly of a replication-competent PCNA-pol δ holoenzyme.

  17. Management of umbilical cord clamping.

    PubMed

    Webbon, Lucy

    2013-02-01

    The Royal College of Midwives (RCM) has updated its third stage of labour guidelines (RCM 2012) to be clearly supportive of a delay in umbilical cord clamping, although specific guidance on timing is yet to be announced. It is therefore imperative that both midwives and student midwives understand and are able to integrate delaying into their practice, as well as communicating to women the benefits; only in this way can we give women fully informed choices on this aspect of care. The main benefit of delayed cord clamping is the protection it can provide in reducing childhood anaemia, which is a major issue, especially in poorer countries. A review of the evidence found no risks linked to delayed clamping, and no evidence that it cannot be used in combination with the administration of uterotonic drugs. Delayed cord clamping can be especially beneficial for pre term and compromised babies.

  18. Theoretical investigation of gain-clamped semiconductor optical amplifiers using the amplified spontaneous emission compensating effect

    NASA Astrophysics Data System (ADS)

    Jia, Xin-Hong

    2006-12-01

    The theoretical model on gain-clamped semiconductor optical amplifiers (GC-SOAs) based on compensating light has been constructed. Using this model, the effects of insertion position and peak reflectivity of the fiber Bragg grating (FBG) on the gain clamping and noise figure (NF) characteristics of GC-SOA are analyzed. The results show that the effect of the FBG insertion position on gain clamping is slight, but the lower NF can be obtained for input FBG-type GC-SOA; when the FBG peak wavelength is designed to close the signal wavelength, the gain clamping and NF characteristics that can be reached are better. Further study shows that, with the increased peak reflectivity of the FBG, the critical input power is broadened and the gain tends to be varied slowly; the larger bias current is helpful to raise gain and decrease the noise figure but is harmful to a gain flatness characteristic.

  19. ACQ4: an open-source software platform for data acquisition and analysis in neurophysiology research.

    PubMed

    Campagnola, Luke; Kratz, Megan B; Manis, Paul B

    2014-01-01

    The complexity of modern neurophysiology experiments requires specialized software to coordinate multiple acquisition devices and analyze the collected data. We have developed ACQ4, an open-source software platform for performing data acquisition and analysis in experimental neurophysiology. This software integrates the tasks of acquiring, managing, and analyzing experimental data. ACQ4 has been used primarily for standard patch-clamp electrophysiology, laser scanning photostimulation, multiphoton microscopy, intrinsic imaging, and calcium imaging. The system is highly modular, which facilitates the addition of new devices and functionality. The modules included with ACQ4 provide for rapid construction of acquisition protocols, live video display, and customizable analysis tools. Position-aware data collection allows automated construction of image mosaics and registration of images with 3-dimensional anatomical atlases. ACQ4 uses free and open-source tools including Python, NumPy/SciPy for numerical computation, PyQt for the user interface, and PyQtGraph for scientific graphics. Supported hardware includes cameras, patch clamp amplifiers, scanning mirrors, lasers, shutters, Pockels cells, motorized stages, and more. ACQ4 is available for download at http://www.acq4.org.

  20. Systematic deletion of homeobox genes in Podospora anserina uncovers their roles in shaping the fruiting body.

    PubMed

    Coppin, Evelyne; Berteaux-Lecellier, Véronique; Bidard, Frédérique; Brun, Sylvain; Ruprich-Robert, Gwenaël; Espagne, Eric; Aït-Benkhali, Jinane; Goarin, Anne; Nesseir, Audrey; Planamente, Sara; Debuchy, Robert; Silar, Philippe

    2012-01-01

    Higher fungi, which comprise ascomycetes and basidiomycetes, play major roles in the biosphere. Their evolutionary success may be due to the extended dikaryotic stage of their life cycle, which is the basis for their scientific name: the Dikarya. Dikaryosis is maintained by similar structures, the clamp in basidiomycetes and the crozier in ascomycetes. Homeodomain transcription factors are required for clamp formation in all basidiomycetes studied. We identified all the homeobox genes in the filamentous ascomycete fungus Podospora anserina and constructed deletion mutants for each of these genes and for a number of gene combinations. Croziers developed normally in these mutants, including those with up to six deleted homeogenes. However, some mutants had defects in maturation of the fruiting body, an effect that could be rescued by providing wild-type maternal hyphae. Analysis of mutants deficient in multiple homeogenes revealed interactions between the genes, suggesting that they operate as a complex network. Similar to their role in animals and plants, homeodomain transcription factors in ascomycetes are involved in shaping multicellular structures.

  1. Systematic Deletion of Homeobox Genes in Podospora anserina Uncovers Their Roles in Shaping the Fruiting Body

    PubMed Central

    Coppin, Evelyne; Berteaux-Lecellier, Véronique; Bidard, Frédérique; Brun, Sylvain; Ruprich-Robert, Gwenaël; Espagne, Eric; Aït-Benkhali, Jinane; Goarin, Anne; Nesseir, Audrey; Planamente, Sara; Debuchy, Robert; Silar, Philippe

    2012-01-01

    Higher fungi, which comprise ascomycetes and basidiomycetes, play major roles in the biosphere. Their evolutionary success may be due to the extended dikaryotic stage of their life cycle, which is the basis for their scientific name: the Dikarya. Dikaryosis is maintained by similar structures, the clamp in basidiomycetes and the crozier in ascomycetes. Homeodomain transcription factors are required for clamp formation in all basidiomycetes studied. We identified all the homeobox genes in the filamentous ascomycete fungus Podospora anserina and constructed deletion mutants for each of these genes and for a number of gene combinations. Croziers developed normally in these mutants, including those with up to six deleted homeogenes. However, some mutants had defects in maturation of the fruiting body, an effect that could be rescued by providing wild-type maternal hyphae. Analysis of mutants deficient in multiple homeogenes revealed interactions between the genes, suggesting that they operate as a complex network. Similar to their role in animals and plants, homeodomain transcription factors in ascomycetes are involved in shaping multicellular structures. PMID:22662159

  2. Weighted augmented Jacobian matrix with a variable coefficient method for kinematics mapping of space teleoperation based on human-robot motion similarity

    NASA Astrophysics Data System (ADS)

    Shi, Zhong; Huang, Xuexiang; Hu, Tianjian; Tan, Qian; Hou, Yuzhuo

    2016-10-01

    Space teleoperation is an important space technology, and human-robot motion similarity can improve the flexibility and intuition of space teleoperation. This paper aims to obtain an appropriate kinematics mapping method of coupled Cartesian-joint space for space teleoperation. First, the coupled Cartesian-joint similarity principles concerning kinematics differences are defined. Then, a novel weighted augmented Jacobian matrix with a variable coefficient (WAJM-VC) method for kinematics mapping is proposed. The Jacobian matrix is augmented to achieve a global similarity of human-robot motion. A clamping weighted least norm scheme is introduced to achieve local optimizations, and the operating ratio coefficient is variable to pursue similarity in the elbow joint. Similarity in Cartesian space and the property of joint constraint satisfaction is analysed to determine the damping factor and clamping velocity. Finally, a teleoperation system based on human motion capture is established, and the experimental results indicate that the proposed WAJM-VC method can improve the flexibility and intuition of space teleoperation to complete complex space tasks.

  3. Simultaneous recording of the action potential and its whole-cell associated ion current on NG108-15 cells cultured over a MWCNT electrode

    NASA Astrophysics Data System (ADS)

    Morales-Reyes, I.; Seseña-Rubfiaro, A.; Acosta-García, M. C.; Batina, N.; Godínez-Fernández, R.

    2016-08-01

    It is well known that, in excitable cells, the dynamics of the ion currents (I i) is extremely important to determine both the magnitude and time course of an action potential (A p). To observe these two processes simultaneously, we cultured NG108-15 cells over a multi-walled carbon nanotubes electrode (MWCNTe) surface and arranged a two independent Patch Clamp system configuration (Bi-Patch Clamp). The first system was used in the voltage or current clamp mode, using a glass micropipette as an electrode. The second system was modified to connect the MWCNTe to virtual ground. While the A p was recorded through the micropipette electrode, the MWCNTe was used to measure the underlying whole-cell current. This configuration allowed us to record both the membrane voltage (V m) and the current changes simultaneously. Images acquired by atomic force microscopy (AFM) and scanning electron microscopy (SEM) indicate that cultured cells developed a complex network of neurites, which served to establish the necessary close contact and strong adhesion to the MWCNTe surface. These features were a key factor to obtain the recording of the whole-cell I i with a high signal to noise ratio (SNR). The experimental results were satisfactorily reproduced by a theoretical model developed to simulate the proposed system. Besides the contribution to a better understanding of the fundamental mechanisms involved in cell communication, the developed method could be useful in cell physiology studies, pharmacology and diseases diagnosis.

  4. Rate dependency of delayed rectifier currents during the guinea-pig ventricular action potential

    PubMed Central

    Rocchetti, Marcella; Besana, Alessandra; Gurrola, Georgina B; Possani, Lourival D; Zaza, Antonio

    2001-01-01

    The action potential clamp technique was exploited to evaluate the rate dependency of delayed rectifier currents (IKr and IKs) during physiological electrical activity. IKr and IKs were measured in guinea-pig ventricular myocytes at pacing cycle lengths (CL) of 1000 and 250 ms.A shorter CL, with the attendant changes in action potential shape, was associated with earlier activation and increased magnitude of both IKr and IKs. Nonetheless, the relative contributions of IKr and IKs to total transmembrane current were independent of CL.Shortening of diastolic interval only (constant action potential shape) enhanced IKs, but not IKr.IKr was increased by a change in the action potential shape only (constant diastolic interval).In ramp clamp experiments, IKr amplitude was directly proportional to repolarization rate at values within the low physiological range (< 1.0 V s−1); at higher repolarization rates proportionality became shallower and finally reversed.When action potential duration (APD) was modulated by constant current injection (I-clamp), repolarization rates > 1.0 V s−1 were associated with a reduced effect of IKr block on APD. The effect of changes in repolarization rate was independent of CL and occurred in the presence of IKs blockade.In spite of its complexity, the behaviour of IKr was accurately predicted by a numerical model based entirely on known kinetic properties of the current.Both IKr and IKs may be increased at fast heart rates, but this may occur through completely different mechanisms. The mechanisms identified are such as to contribute to abnormal rate dependency of repolarization in prolonged repolarization syndromes. PMID:11483703

  5. MATLAB-based automated patch-clamp system for awake behaving mice

    PubMed Central

    Siegel, Jennifer J.; Taylor, William; Chitwood, Raymond A.; Johnston, Daniel

    2015-01-01

    Automation has been an important part of biomedical research for decades, and the use of automated and robotic systems is now standard for such tasks as DNA sequencing, microfluidics, and high-throughput screening. Recently, Kodandaramaiah and colleagues (Nat Methods 9: 585–587, 2012) demonstrated, using anesthetized animals, the feasibility of automating blind patch-clamp recordings in vivo. Blind patch is a good target for automation because it is a complex yet highly stereotyped process that revolves around analysis of a single signal (electrode impedance) and movement along a single axis. Here, we introduce an automated system for blind patch-clamp recordings from awake, head-fixed mice running on a wheel. In its design, we were guided by 3 requirements: easy-to-use and easy-to-modify software; seamless integration of behavioral equipment; and efficient use of time. The resulting system employs equipment that is standard for patch recording rigs, moderately priced, or simple to make. It is written entirely in MATLAB, a programming environment that has an enormous user base in the neuroscience community and many available resources for analysis and instrument control. Using this system, we obtained 19 whole cell patch recordings from neurons in the prefrontal cortex of awake mice, aged 8–9 wk. Successful recordings had series resistances that averaged 52 ± 4 MΩ and required 5.7 ± 0.6 attempts to obtain. These numbers are comparable with those of experienced electrophysiologists working manually, and this system, written in a simple and familiar language, will be useful to many cellular electrophysiologists who wish to study awake behaving mice. PMID:26084901

  6. The Nano-Patch-Clamp Array: Microfabricated Glass Chips for High-Throughput Electrophysiology

    NASA Astrophysics Data System (ADS)

    Fertig, Niels

    2003-03-01

    Electrophysiology (i.e. patch clamping) remains the gold standard for pharmacological testing of putative ion channel active drugs (ICADs), but suffers from low throughput. A new ion channel screening technology based on microfabricated glass chip devices will be presented. The glass chips contain very fine apertures, which are used for whole-cell voltage clamp recordings as well as single channel recordings from mammalian cell lines. Chips containing multiple patch clamp wells will be used in a first bench-top device, which will allow perfusion and electrical readout of each well. This scalable technology will allow for automated, rapid and parallel screening on ion channel drug targets.

  7. Surface characterization of selected LDEF tray clamps

    NASA Technical Reports Server (NTRS)

    Cromer, T. F.; Grammer, H. L.; Wightman, J. P.; Young, Philip R.; Slemp, Wayne S.

    1993-01-01

    The surface characterization of chromic acid anodized 6061-T6 aluminum alloy tray clamps has shown differences in surface chemistry depending upon the position on the Long Duration Exposure Facility (LDEF). Water contact angle results showed no changes in wettability of the tray clamps. The overall surface topography of the control, trailing edge(E3) and leading edge(D9) samples was similar. The thickness of the aluminum oxide layer for all samples determined by Auger depth profiling was less than one micron. X-ray photoelectron spectroscopy (XPS) analysis of the tray clamps showed significant differences in the surface composition. Carbon and silicon containing compounds were the primary contaminants detected.

  8. Implementation of a fast 16-Bit dynamic clamp using LabVIEW-RT.

    PubMed

    Kullmann, Paul H M; Wheeler, Diek W; Beacom, Joshua; Horn, John P

    2004-01-01

    The dynamic-clamp method provides a powerful electrophysiological tool for creating virtual ionic conductances in living cells and studying their influence on membrane potential. Here we describe G-clamp, a new way to implement a dynamic clamp using the real-time version of the Lab-VIEW programming environment together with a Windows host, an embedded microprocessor that runs a real-time operating system and a multifunction data-acquisition board. The software includes descriptions of a fast voltage-dependent sodium conductance, delayed rectifier, M-type and A-type potassium conductances, and a leak conductance. The system can also read synaptic conductance waveforms from preassembled data files. These virtual conductances can be reliably implemented at speeds < or =43 kHz while simultaneously saving two channels of data with 16-bit precision. G-clamp also includes utilities for measuring current-voltage relations, synaptic strength, and synaptic gain. Taking an approach built on a commercially available software/hardware platform has resulted in a system that is easy to assemble and upgrade. In addition, the graphical programming structure of LabVIEW should make it relatively easy for others to adapt G-clamp for new experimental applications.

  9. Dynamic Clamp in Cardiac and Neuronal Systems Using RTXI

    PubMed Central

    Ortega, Francis A.; Butera, Robert J.; Christini, David J.; White, John A.; Dorval, Alan D.

    2016-01-01

    The injection of computer-simulated conductances through the dynamic clamp technique has allowed researchers to probe the intercellular and intracellular dynamics of cardiac and neuronal systems with great precision. By coupling computational models to biological systems, dynamic clamp has become a proven tool in electrophysiology with many applications, such as generating hybrid networks in neurons or simulating channelopathies in cardiomyocytes. While its applications are broad, the approach is straightforward: synthesizing traditional patch clamp, computational modeling, and closed-loop feedback control to simulate a cellular conductance. Here, we present two example applications: artificial blocking of the inward rectifier potassium current in a cardiomyocyte and coupling of a biological neuron to a virtual neuron through a virtual synapse. The design and implementation of the necessary software to administer these dynamic clamp experiments can be difficult. In this chapter, we provide an overview of designing and implementing a dynamic clamp experiment using the Real-Time eXperiment Interface (RTXI), an open- source software system tailored for real-time biological experiments. We present two ways to achieve this using RTXI’s modular format, through the creation of a custom user-made module and through existing modules found in RTXI’s online library. PMID:25023319

  10. Large-Scale Simulation Network Design Study

    DTIC Science & Technology

    1983-10-01

    video displays: three for the tank commander, three for the driver, one for the loader, and one for the gunner. The solid angles subtended by these...Newman Inc Range Sortr This process sorts the expanded display lists into range order for drawing according to the "painter’s algorithm’" The range sorter ...session could then be continued as soon as the network recovered. and the elapsed session time would not be wasted . The SimNet design is much more tolerant

  11. Study and Evaluation of Current and Future Aircraft Loaders

    DTIC Science & Technology

    1986-08-01

    Tigers Airlines JFK Airport , NY 10 Feb 86 Boeing Military Airplane Co. Boeing Commercial Airplane Co. Seattle, WA 11 Feb 86 Korean Airlines LAX, CA 12...Nash Boeing Aircraft Co., Seattle, WA Mr. J. Nelson JFK Airport , NY Mr. P. O’Brien ASD/ENCA Wright-Patterson AFB, OH Mr. C. Outran HQ AFLC/DSTMA...Locations 15 Oct 85 TWA Freight Terminal Laimbert Int’l Airport , St. Louis, mO 22 Nov 85 Lockheed - Georgia Aircraft Co. Marietta, GA 13 Dec 85 Flying

  12. State of the Art for Design and Construction of Sand Compaction Piles

    DTIC Science & Technology

    1987-11-01

    Walz, Headquarters, US Army Corps of Engineers (HQUSACE), was REMR Technical Monitor. The REMR Overview Committee, consisted of Mr. John R. Nikel ... Wire Vibrator Hopper Casing Pipe Air Line Power Line Sand Skip Bucket Front End Loader Figure 2. Typical equipment used to construct a sand...8217~~~~--­ Rubber Packing Wire Inlet for air to close valve and press sand down ..,...__ Air lnl~ot: Figure 5. Special valve used to seal the casing when

  13. Safety Analysis of X4 Multi-Tooled Iowa Detonator Loader

    DTIC Science & Technology

    1980-10-01

    D atmosphere. Temperature identification number should be T5.* Personnel Injury Resulting From Machine Incident - Due to the presence of the barricade...ling factor is the yield strength of velostat . 18 the expectancy is 6 x i0-15 accidents per facility hour. Although the spoon is made of phenolic...expression becoms: I- SF (C-2) v (Cv•cp) (xs) (CV,) 2- uthere: C - coefficient of v4riation for the proess ptent’.4 CV% c"of•iclent of variation "or

  14. Manufactured Soil Field Demonstration for Constructing Wetlands to Treat Acid Mine Drainage on Abandoned Minelands

    DTIC Science & Technology

    2007-11-01

    reclamation and reuse of dredged material from existing CDFs. CRDAs have been established with Recycled Soil Manufacturing Technology (RSMT), N-Viro...existing coal mine bony residual waste. Sawdust was loaded into a manure spreader with a front-end loader and spread across the bony residual in...manure spreader and then dredged material was placed on top of the paper fiber. Both 5 ERDC TN-DOER-D9 November 2007 materials were then spread

  15. Notes on Fire on Aeroplanes

    DTIC Science & Technology

    1917-08-01

    To extract a cartridge or a cartridge case, put the rammer in the muzzle of gun and knock with the head of the rammer on the fuse of the projectile...or on the base of the case. If it is a cartridge which is being extracted ! the loader, standing at the breech, receives it in both hands. (f) Give...fuses and charges. They should be tried in the loading position (incline the gun slightly, open the breech gently, and extract the cartridge with

  16. The dynamic natures of implant loading.

    PubMed

    Wang, Rui-Feng; Kang, Byungsik; Lang, Lisa A; Razzoog, Michael E

    2009-06-01

    A fundamental problem in fully understanding the dynamic nature of implant loading is the confusion that exists regarding the torque load delivered to the implant complex, the initial force transformation/stress/strain developed within the system during the implant complex assembly, and how the clamping forces at the interfaces and the preload stress impact the implant prior to any external loading. The purpose of this study was to create an accurately dimensioned finite element model with spiral threads and threaded bores included in the implant complex, positioned in a bone model, and to determine the magnitude and distribution of the force transformation/stress/strain patterns developed in the modeled implant system and bone and, thus, provide the foundational data for the study of the dynamic loading of dental implants prior to any external loading. An implant (Brånemark Mark III), abutment (CeraOne), abutment screw (Unigrip), and the bone surrounding the implant were modeled using HyperMesh software. The threaded interfaces between screw/implant and implant/bone were designed as a spiral thread helix assigned with specific coefficient of friction values. Assembly simulation using ABAQUS and LS-DYNA was accomplished by applying a 32-Ncm horizontal torque load on the abutment screw (Step 1), then decreasing the torque load to 0 Ncm to simulate the wrench removal (Step 2). The postscript data were collected and reviewed by HyperMesh. A regression analysis was used to depict the relationships between the torque load and the mechanical parameters. During the 32-Ncm tightening sequence, the abutment screw elongated 13.3 mum. The tightening torque generated a 554-N clamping force at the abutment/implant interface and a 522-N preload. The von Mises stress values were 248 MPa in the abutment at the abutment-implant interface, 765 MPa at the top of the screw shaft, 694 MPa at the bottom of the screw shaft, 1365 MPa in the top screw thread, and 21 MPa in the bone at the top of the implant-bone interface. This study also identified various characteristic isosurface stress patterns. The maximum stress magnitude to complete the von Mises stress joint pattern in the present model was 107 MPa during screw tightening, and was reduced to 104 MPa with removal of the wrench. Various specific stress patterns were identified within all elements of the implant complex during the assembly simulation. During the torque moment application, the abutment screw was elongated, and every 1.0-mum elongation of the screw was equivalent to a 47.9-N increase of the preload in the implant complex. The ideal index to determine the preload amount was the contact force at the interface between the screw threads and the threaded screw bore. The isosurface mode identified various characteristic stress patterns developed within the implant complex at the various interfaces during the assembly simulation. These patterns are the (1) spiral and ying-yang pattern of the XY stress, (2) spring, cap, clamping, and preload pattern of the ZZ stress, and (3) bone holding and joint pattern of the von Mises stress.

  17. Characterization of bulbospongiosus muscle reflexes activated by urethral distension in male rats.

    PubMed

    Tanahashi, Masayuki; Karicheti, Venkateswarlu; Thor, Karl B; Marson, Lesley

    2012-10-01

    The urethrogenital reflex (UGR) is used as a surrogate model of the autonomic and somatic nerve and muscle activity that accompanies ejaculation. The UGR is evoked by distension of the urethra and activation of penile afferents. The current study compares two methods of elevating urethral intraluminal pressure in spinalized, anesthetized male Sprague-Dawley rats (n = 60). The first method, penile extension UGR, involves extracting the penis from the foreskin, so that urethral pressure rises due to a natural anatomical flexure in the penis. The second method, penile clamping UGR, involves penile extension UGR with the addition of clamping of the glans penis. Groups of animals were prepared that either received no additional treatment, surgical shams, or received bilateral nerve cuts (4 nerve cut groups): either the pudendal sensory nerve branch (SbPN), the pelvic nerves, the hypogastric nerves, or all three nerves. Penile clamping UGR was characterized by multiple bursts, monitored by electromyography (EMG) of the bulbospongiosus muscle (BSM) accompanied by elevations in urethral pressure. The penile clamping UGR activity declined across multiple trials and eventually resulted in only a single BSM burst, indicating desensitization. In contrast, the penile extension UGR, without penile clamping, evoked only a single BSM EMG burst that showed no desensitization. Thus, the UGR is composed of two BSM patterns: an initial single burst, termed urethrobulbospongiosus (UBS) reflex and a subsequent multiple bursting pattern (termed ejaculation-like response, ELR) that was only induced with penile clamping urethral occlusion. Transection of the SbPN eliminated the ELR in the penile clamping model, but the single UBS reflex remained in both the clamping and extension models. Pelvic nerve (PelN) transection increased the threshold for inducing BSM activation with both methods of occlusion but actually unmasked an ELR in the penile extension method. Hypogastric nerve (HgN) cuts did not significantly alter any parameter. Transection of all three nerves eliminated BSM activation completely. In conclusion, penile clamping occlusion recruits penile and urethral primary afferent fibers that are necessary for an ELR. Urethral distension without significant penile afferent activation recruits urethral primary afferent fibers carried in either the pelvic or pudendal nerve that are necessary for the single-burst UBS reflex.

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

    Mays, Suzanne G.; Okafor, C. Denise; Tuntland, Micheal L.

    Peroxisome proliferator-activated gamma coactivator 1-α (PGC1α) regulates energy metabolism by directly interacting with transcription factors to modulate gene expression. Among the PGC1α binding partners is liver receptor homolog 1 (LRH-1; NR5A2), an orphan nuclear hormone receptor that controls lipid and glucose homeostasis. Although PGC1α is known to bind and activate LRH-1, mechanisms through which PGC1α changes LRH-1 conformation to drive transcription are unknown. Here, we used biochemical and structural methods to interrogate the LRH-1–PGC1α complex. Purified, full-length LRH-1, as well as isolated ligand binding domain, bound to PGC1α with higher affinity than to the coactivator, nuclear receptor coactivator-2 (Tif2), inmore » coregulator peptide recruitment assays. We present the first crystal structure of the LRH-1–PGC1α complex, which depicts several hydrophobic contacts and a strong charge clamp at the interface between these partners. In molecular dynamics simulations, PGC1α induced correlated atomic motion throughout the entire LRH-1 activation function surface, which was dependent on charge-clamp formation. In contrast, Tif2 induced weaker signaling at the activation function surface than PGC1α but promoted allosteric signaling from the helix 6/β-sheet region of LRH-1 to the activation function surface. These studies are the first to probe mechanisms underlying the LRH-1–PGC1α interaction and may illuminate strategies for selective therapeutic targeting of PGC1α-dependent LRH-1 signaling pathways.« less

  19. Product Development and Cost Analysis of Fabricating the Prototype of Roller Clamp in Intravenous (I.V) Tubing Medical Devices using Fused Deposition Modeling (FDM) Technology

    NASA Astrophysics Data System (ADS)

    Way, Yusoff

    2018-01-01

    The main aim of this research is to develop a new prototype and to conduct cost analysis of the existing roller clamp which is one of parts attached to Intravenous (I.V) Tubing used in Intravenous therapy medical device. Before proceed with the process to manufacture the final product using Fused Deposition Modeling (FDM) Technology, the data collected from survey were analyzed using Product Design Specifications approach. Selected concept has been proven to have better quality, functions and criteria compared to the existing roller clamp and the cost analysis of fabricating the roller clamp prototype was calculated.

  20. Enhanced strain effect of aged acceptor-doped BaTiO3 ceramics with clamping domain structures

    NASA Astrophysics Data System (ADS)

    Wang, Lei; Zhou, Zhiyong; Zhao, Xiaobo; Liu, Zhen; Liang, Ruihong; Dong, Xianlin

    2017-03-01

    A clamping domain structure is proposed to improve the amount of non-180° domain switching in BaTiO3 based piezoelectric ceramics. Experimental results show a large unipolar strain of 0.23% at 5 kV/mm in aged 0.5 mol. % Mn doped BaTiO3 ceramics with clamping domain structures, and the normalized strain (d33*= Smax/Emax) reaches 600 pm/V at low electric fields of 2 or 3 kV/mm. In contrast, pure BaTiO3 ceramics with clamping domain structures exhibit no clear polarization constriction or strain enhancement at 3 kV/mm. Electron paramagnetic resonance spectra verify the existence of titanium vacancies, Mn2+ and Mn4+, in 0.5 mol. % Mn doped BaTiO3 ceramics. These results indicate that the enhanced strain effect can be attributed to the combined effect of the clamping domain structure and stabilization of defect dipoles. This method provides a general way to obtain large strain in ferroelectrics.

  1. Service equipment for use in hostile environments

    NASA Technical Reports Server (NTRS)

    Dolce, James L. (Inventor); Gordon, Andrew L. (Inventor)

    1994-01-01

    Service equipment for use in hostile environments includes a detachable service unit secured to a stationary service unit. The detachable service unit includes a housing with an exterior plate, a power control interface for connection to an exterior power source, locating pins located in said exterior plate, an electrical connector in the exterior plate electrically coupled to said power control interface, and a pair of clamping receptacles formed in the exterior plate and located on adjacent opposite edges of the exterior plate. The stationary unit includes an electrical connector for connection to the electrical connector of the detachable service unit, a clamping apparatus for clamping and unclamping the detachable service unit from the stationary unit, a base clamp assembly for mounting the clamping apparatus onto the stationary unit, and locating pin holes for receiving the locating pins and aligning the detachable service unit onto the stationary unit. The detachable service unit and stationary unit have mating scalloped faces which aid in alignment and provide a mechanism for heat dissipation.

  2. Robotic Automation of In Vivo Two-Photon Targeted Whole-Cell Patch-Clamp Electrophysiology.

    PubMed

    Annecchino, Luca A; Morris, Alexander R; Copeland, Caroline S; Agabi, Oshiorenoya E; Chadderton, Paul; Schultz, Simon R

    2017-08-30

    Whole-cell patch-clamp electrophysiological recording is a powerful technique for studying cellular function. While in vivo patch-clamp recording has recently benefited from automation, it is normally performed "blind," meaning that throughput for sampling some genetically or morphologically defined cell types is unacceptably low. One solution to this problem is to use two-photon microscopy to target fluorescently labeled neurons. Combining this with robotic automation is difficult, however, as micropipette penetration induces tissue deformation, moving target cells from their initial location. Here we describe a platform for automated two-photon targeted patch-clamp recording, which solves this problem by making use of a closed loop visual servo algorithm. Our system keeps the target cell in focus while iteratively adjusting the pipette approach trajectory to compensate for tissue motion. We demonstrate platform validation with patch-clamp recordings from a variety of cells in the mouse neocortex and cerebellum. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  3. Industrializing electrophysiology: HT automated patch clamp on SyncroPatch® 96 using instant frozen cells.

    PubMed

    Polonchuk, Liudmila

    2014-01-01

    Patch-clamping is a powerful technique for investigating the ion channel function and regulation. However, its low throughput hampered profiling of large compound series in early drug development. Fortunately, automation has revolutionized the area of experimental electrophysiology over the past decade. Whereas the first automated patch-clamp instruments using the planar patch-clamp technology demonstrated rather a moderate throughput, few second-generation automated platforms recently launched by various companies have significantly increased ability to form a high number of high-resistance seals. Among them is SyncroPatch(®) 96 (Nanion Technologies GmbH, Munich, Germany), a fully automated giga-seal patch-clamp system with the highest throughput on the market. By recording from up to 96 cells simultaneously, the SyncroPatch(®) 96 allows to substantially increase throughput without compromising data quality. This chapter describes features of the innovative automated electrophysiology system and protocols used for a successful transfer of the established hERG assay to this high-throughput automated platform.

  4. Patch-clamp amplifiers on a chip

    PubMed Central

    Weerakoon, Pujitha; Culurciello, Eugenio; Yang, Youshan; Santos-Sacchi, Joseph; Kindlmann, Peter J.; Sigworth, Fred J.

    2010-01-01

    We present the first, fully-integrated, two-channel implementation of a patch-clamp measurement system. With this “PatchChip” two simultaneous whole-cell recordings can be obtained with rms noise of 8 pA in a 10 kHz bandwidth. The capacitance and series-resistance of the electrode can be compensated up to 10 pF and 100 MΩ respectively under computer control. Recordings of hERG and Nav 1.7 currents demonstrate the system's capabilities, which are on par with large, commercial patch-clamp instrumentation. By reducing patch-clamp amplifiers to a millimeter size micro-chip, this work paves the way to the realization of massively-parallel, high-throughput patch-clamp systems for drug screening and ion-channel research. The PatchChip is implemented in a 0.5 μm silicon-on-sapphire process; its size is 3 × 3 mm2 and the power consumption is 5 mW per channel with a 3.3 V power supply. PMID:20637803

  5. Conformational analysis of processivity clamps in solution demonstrates that tertiary structure does not correlate with protein dynamics.

    PubMed

    Fang, Jing; Nevin, Philip; Kairys, Visvaldas; Venclovas, Česlovas; Engen, John R; Beuning, Penny J

    2014-04-08

    The relationship between protein sequence, structure, and dynamics has been elusive. Here, we report a comprehensive analysis using an in-solution experimental approach to study how the conservation of tertiary structure correlates with protein dynamics. Hydrogen exchange measurements of eight processivity clamp proteins from different species revealed that, despite highly similar three-dimensional structures, clamp proteins display a wide range of dynamic behavior. Differences were apparent both for structurally similar domains within proteins and for corresponding domains of different proteins. Several of the clamps contained regions that underwent local unfolding with different half-lives. We also observed a conserved pattern of alternating dynamics of the α helices lining the inner pore of the clamps as well as a correlation between dynamics and the number of salt bridges in these α helices. Our observations reveal that tertiary structure and dynamics are not directly correlated and that primary structure plays an important role in dynamics. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Influence of clamp-up force on the strength of bolted composite joints

    NASA Astrophysics Data System (ADS)

    Horn, Walter J.; Schmitt, Ron R.

    1994-03-01

    Composite materials offer the potential for a reduction in the number of individual parts and joints in a structure because large one-piece components can replace multipart assemblies. Nevertheless, there are many situations where composite parts must be joined and often mechanical fasteners provide the only practical method of joining those parts. The long-term strength of mechanically fastened joints of composite members can be directly affected by the clamp-up force of the fastener and thus perhaps by the relaxation of this force due to the viscoelastic character of the composite materials of the joint. Methods for predicting the effect of bolt clamp-up force relaxation on the strength of mechanically fastened joints of thermoplastic composite materials were investigated during the present study. A test program, using two thermoplastic composite materials, was conducted to determine the influence of clamp-up force on joint strength, to measure the relaxation of the joint clamp-up force with time, and to measure the change of joint strength as a function of time.

  7. Experimental and numerical analysis of clamped joints in front motorbike suspensions

    NASA Astrophysics Data System (ADS)

    Croccolo, D.; de Agostinis, M.; Vincenzi, N.

    2010-06-01

    Clamped joints are shaft-hub connections used, as an instance, in front motorbike suspensions to lock the steering plates with the legs and the legs with the wheel pin, by means of one or two bolts. The preloading force, produced during the tightening process, should be evaluated accurately, since it must lock safely the shaft, without overcoming the yielding point of the hub. Firstly, friction coefficients have been evaluated on “ad-hoc designed” specimens, by applying the Design of Experiment approach: the applied tightening torque has been precisely related to the imposed preloading force. Then, the tensile state of clamps have been evaluated both via FEM and by leveraging some design formulae proposed by the Authors as function of the preloading force and of the clamp geometry. Finally, the results have been compared to those given by some strain gauges applied on the tested clamps: the discrepancies between numerical analyses, the design formulae and the experimental results remains under a threshold of 10%.

  8. The Fibroblast Growth Factor 14·Voltage-gated Sodium Channel Complex Is a New Target of Glycogen Synthase Kinase 3 (GSK3)*

    PubMed Central

    Shavkunov, Alexander S.; Wildburger, Norelle C.; Nenov, Miroslav N.; James, Thomas F.; Buzhdygan, Tetyana P.; Panova-Elektronova, Neli I.; Green, Thomas A.; Veselenak, Ronald L.; Bourne, Nigel; Laezza, Fernanda

    2013-01-01

    The FGF14 protein controls biophysical properties and subcellular distribution of neuronal voltage-gated Na+ (Nav) channels through direct binding to the channel C terminus. To gain insights into the dynamic regulation of this protein/protein interaction complex, we employed the split luciferase complementation assay to screen a small molecule library of kinase inhibitors against the FGF14·Nav1.6 channel complex and identified inhibitors of GSK3 as hits. Through a combination of a luminescence-based counter-screening, co-immunoprecipitation, patch clamp electrophysiology, and quantitative confocal immunofluorescence, we demonstrate that inhibition of GSK3 reduces the assembly of the FGF14·Nav channel complex, modifies FGF14-dependent regulation of Na+ currents, and induces dissociation and subcellular redistribution of the native FGF14·Nav channel complex in hippocampal neurons. These results further emphasize the role of FGF14 as a critical component of the Nav channel macromolecular complex, providing evidence for a novel GSK3-dependent signaling pathway that might control excitability through specific protein/protein interactions. PMID:23640885

  9. Normal Modes of Vibration of the PHALANX Gun

    DTIC Science & Technology

    1993-06-01

    Clamps Bricks, Thin Shells, Rigid Elements Mid-Barrel Clamps Bricks, Rigid Elements Barrels Beams with tubular cross-section Stub Rotor Bricks, Thin...Shells Rotor Bricks Needle Bearing Bricks, Springs Casing Thin Shells Thrust Bearing Bricks, Springs Recoil Adapters Bricks, Rigid Elements, Springs... rigid elements were used to connect the barrels to the clamps and stub rotor and the recoil adapter springs to 48 the gun body. "End release codes

  10. Single mode variable-sensitivity fiber optic sensors

    NASA Technical Reports Server (NTRS)

    Murphy, K. A.; Fogg, B. R.; Gunther, M. F.; Claus, R. O.

    1992-01-01

    We review spatially-weighted optical fiber sensors that filter specific vibration modes from one dimensional beams placed in clamped-free and clamped-clamped configurations. The sensitivity of the sensor is varied along the length of the fiber by tapering circular-core, dual-mode optical fibers. Selective vibration mode suppression on the order of 10 dB was obtained. We describe experimental results and propose future extensions to single mode sensor applications.

  11. Pitfalls during biomechanical testing - Evaluation of different fixation methods for measuring tendons endurance properties.

    PubMed

    Hangody, Gy; Pánics, G; Szebényi, G; Kiss, R; Hangody, L; Pap, K

    2016-03-01

    The goal of the study was to find a proper technique to fix tendon grafts into an INSTRON loading machine. From 8 human cadavers, 40 grafts were collected. We removed the bone-patella tendon-bone grafts, the semitendinosus and gracilis tendons, the quadriceps tendon-bone grafts, the Achilles tendons, and the peroneus longus tendons from each lower extremity. We tested the tendon grafts with five different types of fixation devices: surgical thread (Premicron 3), general mounting clamp, wire mesh, cement fixation, and a modified clamp for an INSTRON loading machine. The mean failure load in case of surgical thread fixation was (381N ± 26N). The results with the general clamp were (527N ± 45N). The wire meshes were more promising (750N ± 21N), but did not reach the outcomes we desired. Easy slippages of the ends of the tendons from the cement encasements were observed (253N ± 18N). We then began to use Shi's clamp that could produce 977N ± 416N peak force. We combined Shi's clamp with freezing of the graft and the rupture of the tendon itself demonstrated an average force of 2198 N ± 773N. We determined that our modified frozen clamp fixed the specimens against high tensile forces.

  12. An accurate online calibration system based on combined clamp-shape coil for high voltage electronic current transformers.

    PubMed

    Li, Zhen-hua; Li, Hong-bin; Zhang, Zhi

    2013-07-01

    Electronic transformers are widely used in power systems because of their wide bandwidth and good transient performance. However, as an emerging technology, the failure rate of electronic transformers is higher than that of traditional transformers. As a result, the calibration period needs to be shortened. Traditional calibration methods require the power of transmission line be cut off, which results in complicated operation and power off loss. This paper proposes an online calibration system which can calibrate electronic current transformers without power off. In this work, the high accuracy standard current transformer and online operation method are the key techniques. Based on the clamp-shape iron-core coil and clamp-shape air-core coil, a combined clamp-shape coil is designed as the standard current transformer. By analyzing the output characteristics of the two coils, the combined clamp-shape coil can achieve verification of the accuracy. So the accuracy of the online calibration system can be guaranteed. Moreover, by employing the earth potential working method and using two insulating rods to connect the combined clamp-shape coil to the high voltage bus, the operation becomes simple and safe. Tests in China National Center for High Voltage Measurement and field experiments show that the proposed system has a high accuracy of up to 0.05 class.

  13. HTS techniques for patch clamp-based ion channel screening - advances and economy.

    PubMed

    Farre, Cecilia; Fertig, Niels

    2012-06-01

    Ten years ago, the first publication appeared showing patch clamp recordings performed on a planar glass chip instead of using a conventional patch clamp pipette. "Going planar" proved to revolutionize ion channel drug screening as we know it, by allowing high quality measurements of ion channels and their effectors at a higher throughput and at the same time de-skilling the highly laborious technique. Over the years, platforms evolved in response to user requirements regarding experimental features, data handling plus storage, and suitable target diversity. This article gives a snapshot image of patch clamp-based ion channel screening with focus on platforms developed to meet requirements of high-throughput screening environments. The commercially available platforms are described, along with their benefits and drawbacks in ion channel drug screening. Automated patch clamp (APC) platforms allow faster investigation of a larger number of ion channel active compounds or cell clones than previously possible. Since patch clamp is the only method allowing direct, real-time measurements of ion channel activity, APC holds the promise of picking up high quality leads, where they otherwise would have been overseen using indirect methods. In addition, drug candidate safety profiling can be performed earlier in the drug discovery process, avoiding late-phase compound withdrawal due to safety liability issues, which is highly costly and inefficient.

  14. Ion channel drug discovery and research: the automated Nano-Patch-Clamp technology.

    PubMed

    Brueggemann, A; George, M; Klau, M; Beckler, M; Steindl, J; Behrends, J C; Fertig, N

    2004-01-01

    Unlike the genomics revolution, which was largely enabled by a single technological advance (high throughput sequencing), rapid advancement in proteomics will require a broader effort to increase the throughput of a number of key tools for functional analysis of different types of proteins. In the case of ion channels -a class of (membrane) proteins of great physiological importance and potential as drug targets- the lack of adequate assay technologies is felt particularly strongly. The available, indirect, high throughput screening methods for ion channels clearly generate insufficient information. The best technology to study ion channel function and screen for compound interaction is the patch clamp technique, but patch clamping suffers from low throughput, which is not acceptable for drug screening. A first step towards a solution is presented here. The nano patch clamp technology, which is based on a planar, microstructured glass chip, enables automatic whole cell patch clamp measurements. The Port-a-Patch is an automated electrophysiology workstation, which uses planar patch clamp chips. This approach enables high quality and high content ion channel and compound evaluation on a one-cell-at-a-time basis. The presented automation of the patch process and its scalability to an array format are the prerequisites for any higher throughput electrophysiology instruments.

  15. Timing of cord clamping in very preterm infants: more evidence is needed.

    PubMed

    Tarnow-Mordi, William O; Duley, Lelia; Field, David; Marlow, Neil; Morris, Jonathan; Newnham, John; Paneth, Nigel; Soll, Roger F; Sweet, David

    2014-08-01

    In December 2012, the American College of Obstetricians and Gynecologists published a Committee Opinion entitled "Timing of umbilical cord clamping after birth." It stated that "evidence exists to support delayed cord clamping in preterm infants, when feasible. The single most important benefit for preterm infants is the possibility for a nearly 50% reduction in IVH." However, the Committee Opinion added that the ideal timing of umbilical cord clamping has yet to be determined and recommended that large clinical trials be conducted in the most preterm infants. Published randomized controlled trials include <200 infants of <30 weeks' gestation, with assessments of neurodevelopmental outcome in less than one-half of the children. This is a major gap in the evidence. Without reliable data from randomized controlled trials that optimally include childhood follow-up evaluations, we will not know whether delayed cord clamping may do more overall harm than good. Ongoing trials of delayed cord clamping plan to report childhood outcomes in >2000 additional very preterm infants. Current recommendations may need to change when these results become available. Greater international collaboration could accelerate resolution of whether this promising intervention will improve disability-free survival in about 1 million infants who will be born very preterm globally each year. Copyright © 2014 Mosby, Inc. All rights reserved.

  16. Glomerular loss after arteriovenous and arterial clamping for renal warm ischemia in a swine model.

    PubMed

    Bechara, Gustavo Ruschi; Damasceno-Ferreira, José Aurelino; Abreu, Leonardo Albuquerque Dos Santos; Costa, Waldemar Silva; Sampaio, Francisco José Barcellos; Pereira-Sampaio, Marco Aurélio; Souza, Diogo Benchimol De

    2016-11-01

    To evaluate the glomerular loss after arteriovenous or arterial warm ischemia in a swine model. Twenty four pigs were divided into Group Sham (submitted to all surgical steps except the renal ischemia), Group AV (submitted to 30 minutes of warm ischemia by arteriovenous clamping of left kidney vessels), and Group A (submitted to 30 minutes of ischemia by arterial clamping). Right kidneys were used as controls. Weigh, volume, cortical volume, glomerular volumetric density (Vv[Glom]), volume-weighted glomerular volume (VWGV), and the total number of glomeruli were measured for each organ. Group AV showed a 24.5% reduction in its left kidney Vv[Glom] and a 25.4% reduction in the VWGV, when compared to the right kidney. Reductions were also observed when compared to kidneys of sham group. There was a reduction of 19.2% in the total number of glomeruli in AV kidneys. No difference was observed in any parameters analyzed on the left kidneys from group A. Renal warm ischemia of 30 minutes by arterial clamping did not caused significant glomerular damage, but arteriovenous clamping caused significant glomerular loss in a swine model. Clamping only the renal artery should be considered to minimize renal injury after partial nephrectomies.

  17. Cerebral watershed infarcts may be induced by hemodynamic changes in blood flow.

    PubMed

    Shi, Jingfei; Meng, Ran; Konakondla, Sanjay; Ding, Yuchuan; Duan, Yunxia; Wu, Di; Wang, Bincheng; Luo, Yinghao; Ji, Xunming

    2017-06-01

    A watershed infarct is defined as an ischemic lesion at the border zones between territories of two major arteries. The pathogenesis of watershed infarcts, specifically whether they are caused by hemodynamic or embolic mechanisms, has long been debated. In this study, we aimed to examine whether watershed infarcts can be induced by altering the hemodynamic conditions in rats. In phase one, to determine the proper clamping duration for a reproducible infarct, 30 rats were equally divided into 5 subgroups and underwent bilateral common carotid artery (CCA) clamping for different durations (0.5, 1.0, 1.5, 2.0, and 3.0 hours). In phase two, to analyze the types of infarcts induced by bilateral CCA clamping, 40 rats were subjected to bilateral CCA clamping for 2 hours. As a control, 8 rats underwent all the operation procedures except bilateral CCA clamping. We performed 7.0T magnetic resonance imaging on the surviving rats on the second day to evaluate the extent of the infarcts. We further identified and examined the infarcts with brain slices stained using 2, 3, 5-triphenyltetrazolium chloride (TTC) on the third day. After 2 hours of bilateral CCA clamping, cerebral infarction occurred in 42% of surviving rats (13/31). The majority of the ischemic lesions were located in watershed regions of the brain, demonstrated by both MRI and TTC staining. Watershed infarcts were induced through changing hemodynamic conditions by bilateral CCA clamping in rats. This method may lead to the development of a reliable rodent model for watershed infarcts.

  18. Patch-Clamp Recording from Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes: Improving Action Potential Characteristics through Dynamic Clamp

    PubMed Central

    Veerman, Christiaan C.; Zegers, Jan G.; Mengarelli, Isabella; Bezzina, Connie R.

    2017-01-01

    Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) hold great promise for studying inherited cardiac arrhythmias and developing drug therapies to treat such arrhythmias. Unfortunately, until now, action potential (AP) measurements in hiPSC-CMs have been hampered by the virtual absence of the inward rectifier potassium current (IK1) in hiPSC-CMs, resulting in spontaneous activity and altered function of various depolarising and repolarising membrane currents. We assessed whether AP measurements in “ventricular-like” and “atrial-like” hiPSC-CMs could be improved through a simple, highly reproducible dynamic clamp approach to provide these cells with a substantial IK1 (computed in real time according to the actual membrane potential and injected through the patch-clamp pipette). APs were measured at 1 Hz using perforated patch-clamp methodology, both in control cells and in cells treated with all-trans retinoic acid (RA) during the differentiation process to increase the number of cells with atrial-like APs. RA-treated hiPSC-CMs displayed shorter APs than control hiPSC-CMs and this phenotype became more prominent upon addition of synthetic IK1 through dynamic clamp. Furthermore, the variability of several AP parameters decreased upon IK1 injection. Computer simulations with models of ventricular-like and atrial-like hiPSC-CMs demonstrated the importance of selecting an appropriate synthetic IK1. In conclusion, the dynamic clamp-based approach of IK1 injection has broad applicability for detailed AP measurements in hiPSC-CMs. PMID:28867785

  19. Catch and Patch: A Pipette-Based Approach for Automating Patch Clamp That Enables Cell Selection and Fast Compound Application.

    PubMed

    Danker, Timm; Braun, Franziska; Silbernagl, Nikole; Guenther, Elke

    2016-03-01

    Manual patch clamp, the gold standard of electrophysiology, represents a powerful and versatile toolbox to stimulate, modulate, and record ion channel activity from membrane fragments and whole cells. The electrophysiological readout can be combined with fluorescent or optogenetic methods and allows for ultrafast solution exchanges using specialized microfluidic tools. A hallmark of manual patch clamp is the intentional selection of individual cells for recording, often an essential prerequisite to generate meaningful data. So far, available automation solutions rely on random cell usage in the closed environment of a chip and thus sacrifice much of this versatility by design. To parallelize and automate the traditional patch clamp technique while perpetuating the full versatility of the method, we developed an approach to automation, which is based on active cell handling and targeted electrode placement rather than on random processes. This is achieved through an automated pipette positioning system, which guides the tips of recording pipettes with micrometer precision to a microfluidic cell handling device. Using a patch pipette array mounted on a conventional micromanipulator, our automated patch clamp process mimics the original manual patch clamp as closely as possible, yet achieving a configuration where recordings are obtained from many patch electrodes in parallel. In addition, our implementation is extensible by design to allow the easy integration of specialized equipment such as ultrafast compound application tools. The resulting system offers fully automated patch clamp on purposely selected cells and combines high-quality gigaseal recordings with solution switching in the millisecond timescale.

  20. Rectal transection using a curved cutter stapler with an endo-Satinsky clamp during a laparoscopic low anterior resection.

    PubMed

    Hotta, Tsukasa; Takifuji, Katsunari; Yokoyama, Shozo; Matsuda, Kenji; Yamaue, Hiroki

    2012-10-01

    A new rectal transaction method was developed using a combination of the curved cutter stapler and endo-Satinsky clamp because of the difficulty in performing rectal transection in the narrow pelvic cavity. The endo-Satinsky clamp is inserted without a flexible trocar cannula by connecting the handle extra-abdominally with a shaft of the endo-Satinsky clamp through the left higher quadrant port via a retrograde course from a midline incision above the pubis symphysis. The endo-Satinsky clamp is used to clamp the rectal wall horizontally at the distal end of the tumor. The wrist of an elastic surgical glove fixed with the shaft of the curved cutter stapler is covered with a midline incision, and consequently, the stapler is inserted into the pelvic cavity. The curved head of the stapler is rotated to the left at the anal side of the endo-Satinsky clamp to insert the rectum between the jaws of the stapler. The stapler is closed and fired, and a rectal transection is thus performed with one firing using a single cartridge. This method was performed in 12 patients with rectal cancer. The median value and range of the tumor distance from the anal verge were 7.0 and 4.5-11.0 cm, respectively. The median duration of the operation was 252 min, and the median blood loss was 15 mL. Only one stapling cartridge was used for rectal transection in all cases, and no major complications were observed. We herein demonstrated a new transection method for rectal cancer.

  1. Factors in daily physical activity related to calcaneal mineral density in men.

    PubMed

    Hutchinson, T M; Whalen, R T; Cleek, T M; Vogel, J M; Arnaud, S B

    1995-05-01

    To determine the factors in daily physical activity that influence the mineral density of the calcaneus, we recorded walking steps and the type and duration of exercise in 43 healthy 26-to 51-yr-old men. Areal (g.cm-2) calcaneal bone mineral density (CBMD) was measured by single energy x-ray densitometry (SXA, Osteon, Inc., Wahiawa, HI). Subjects walked a mean (+/- SD) of 7902 (+/- 2534) steps per day or approximately 3.9 (+/- 1.2) miles daily. Eight subjects reported no exercise activities. The remaining 35 subjects spent 143 (2-772) (median and range) min.wk-1 exercising. Twenty-eight men engaged in exercise activities that generate single leg peak vertical ground reaction forces (GRFz) of 2 or more body weights (high loaders, HL), and 15 reported exercise or daily activities that typically generate GRFz less than 1.5 body weights (low loaders, LL). CBMD was 12% higher in HL than LL (0.668 +/- 0.074 g.cm-2 vs 0.597 +/- 0.062 g.cm-2, P < 0.004). In the HL group, CBMD correlated to reported minutes of high load exercise (r = 0.41, P < 0.03). CBMD was not related to the number of daily walking steps (N = 43, r = 0.03, NS). The results of this study support the concept that the dominant factor in daily physical activity relating to bone mineral density is the participation in site specific high loading activities, i.e., for the calcaneus, high calcaneal loads.

  2. Factors in Daily Physical Activity Related to Calcaneal Mineral Density in Men

    NASA Technical Reports Server (NTRS)

    Hutchinson, Teresa M.; Whalen, Robert T.; Cleek, Tammy M.; Vogel, John M.; Arnaud, Sara B.

    1995-01-01

    To determine the factors in daily physical activity that influence the mineral density of the calcaneus, we recorded walking steps and the type and duration of exercise in 43 healthy 26-to 51-yr-old men. Areal (g/sq cm) calcaneal bone mineral density (CBMD) was measured by single energy x-ray densitometry. Subjects walked a mean (+/- SD) of 7902(+/-2534) steps per day or approximately 3.9(+/-1.2) miles daily. Eight subjects reported no exercise activities. The remaining 35 subjects spent 143(2-772) (median and range) min/wk exercising. Twenty-eight men engaged in exercise activities that generate single leg peak vertical ground reaction forces (GRF(sub z)) of 2 or more body weights (high loaders, HL), and 15 reported exercise or daily activities that typically generate GRF(sub z) less than 1.5 body weights (low loaders, LL). CBMD was 12% higher in HL than LL (0.668 +/- 0.074 g/sq cm vs 0.597 +/- 0.062 g/sq cm, P less than 0.004). In the HL group, CBMD correlated to reported minutes of high load exercise (r = 0.41, P less than 0.03). CBMD was not related to the number of daily walking steps (N = 43, r = 0.03, NS). The results of this study support the concept that the dominant factor in daily physical activity relating to bone mineral density is the participation in site specific high loading activities, i.e., for the calcaneus, high calcaneal loads.

  3. Apoplastic and symplastic phloem loading in Quercus robur and Fraxinus excelsior

    PubMed Central

    Lohaus, Gertrud

    2014-01-01

    Whereas most of the research on phloem loading is performed on herbaceous plants, less is known about phloem loading strategies in trees. In this study, the phloem loading mechanisms of Quercus robur and Fraxinus excelsior were analysed. The following features were examined: the minor vein structure, the sugar concentrations in phloem sap by the laser–aphid–stylet technique, the distribution of photoassimilates in the mesophyll cells by non-aqueous fractionation, gradients of sugar concentrations and osmotic pressure, and the expression of sucrose transporters. The minor vein configurations of Q. robur and F. excelsior belong to the open type. Quercus robur contained companion cells in the minor veins whereas F. excelsior showed intermediary cells in addition to ordinary companion cells. The main carbon transport form in Q. robur was sucrose (~1M). In F. excelsior high amounts of raffinose and stachyose were also transported. However, in both tree species, the osmolality of phloem sap was higher than the osmolality of the mesophyll cells. The concentration gradients between phloem sap and the cytoplasm of mesophyll cells for sucrose were 16-fold and 14-fold for Q. robur and F. excelsior, respectively. Independent of the type of translocated sugars, sucrose transporter cDNAs were cloned from both species. The results indicate that phloem loading of sucrose and other metabolites must involve active loading steps in both tree species. Quercus robur seems to be an apoplastic phloem loader while F. excelsior shows indications of being a symplastic or mixed symplastic–apoplastic phloem loader. PMID:24591056

  4. Muscle-spring dynamics in time-limited, elastic movements.

    PubMed

    Rosario, M V; Sutton, G P; Patek, S N; Sawicki, G S

    2016-09-14

    Muscle contractions that load in-series springs with slow speed over a long duration do maximal work and store the most elastic energy. However, time constraints, such as those experienced during escape and predation behaviours, may prevent animals from achieving maximal force capacity from their muscles during spring-loading. Here, we ask whether animals that have limited time for elastic energy storage operate with springs that are tuned to submaximal force production. To answer this question, we used a dynamic model of a muscle-spring system undergoing a fixed-end contraction, with parameters from a time-limited spring-loader (bullfrog: Lithobates catesbeiana) and a non-time-limited spring-loader (grasshopper: Schistocerca gregaria). We found that when muscles have less time to contract, stored elastic energy is maximized with lower spring stiffness (quantified as spring constant). The spring stiffness measured in bullfrog tendons permitted less elastic energy storage than was predicted by a modelled, maximal muscle contraction. However, when muscle contractions were modelled using biologically relevant loading times for bullfrog jumps (50 ms), tendon stiffness actually maximized elastic energy storage. In contrast, grasshoppers, which are not time limited, exhibited spring stiffness that maximized elastic energy storage when modelled with a maximal muscle contraction. These findings demonstrate the significance of evolutionary variation in tendon and apodeme properties to realistic jumping contexts as well as the importance of considering the effect of muscle dynamics and behavioural constraints on energy storage in muscle-spring systems. © 2016 The Author(s).

  5. Renal functional and perioperative outcomes of off-clamp versus clamped robot-assisted partial nephrectomy: matched cohort study.

    PubMed

    Tanagho, Youssef S; Bhayani, Sam B; Sandhu, Gurdarshan S; Vaughn, Nicholas P; Nepple, Kenneth G; Figenshau, R Sherburne

    2012-10-01

    To evaluate the potential benefit of performing off-clamp robot-assisted partial nephrectomy as it relates to renal functional outcomes, while assessing the safety profile of this unconventional surgical approach. Twenty-nine patients who underwent off-clamp robot-assisted partial nephrectomy for suspected renal cell carcinoma at Washington University between March 2008 and September 2011 (group 1) were matched to 29 patients with identical nephrometry scores and comparable baseline renal function who underwent robot-assisted partial nephrectomy with hilar clamping during the same period (group 2). The matched cohorts' perioperative and renal functional outcomes were compared at a mean 9-month follow-up. Mean estimated blood loss was 146.4 mL in group 1, versus 103.9 mL in group 2 (P = .039). Mean hilar clamp time was 0 minutes in group 1 and 14.7 minutes in group 2. No perioperative complications were encountered in group 1; 1 Clavien-2 complication (3.4%) occurred in group 2 (P = 1.000). At 9-month follow-up, mean estimated glomerular filtration rate in group 1 was 79.9 versus 84.8 mL/min/1.73 m(2) preoperatively (P = .013); mean estimated glomerular filtration rate in group 2 was 74.1 versus 85.8 mL/min/1.73 m(2) preoperatively (P < .001). Hence, estimated glomerular filtration rate declined by a mean of 4.9 mL/min/1.73 m(2) in group 1 versus 11.7 mL/min/1.73 m(2) in group 2 (P = .033). Off-clamp robot-assisted partial nephrectomy is associated with a favorable morbidity profile and relatively greater renal functional preservation compared to clamped robot-assisted partial nephrectomy. Nevertheless, the benefit is small in renal functional terms and may have very limited clinical relevance. Copyright © 2012 Elsevier Inc. All rights reserved.

  6. Timing of umbilical cord-clamping and infant anaemia: the role of maternal anaemia.

    PubMed

    Blouin, Brittany; Penny, Mary E; Maheu-Giroux, Mathieu; Casapía, Martín; Aguilar, Eder; Silva, Hermánn; Creed-Kanashiro, Hilary M; Joseph, Serene A; Gagnon, Anita; Rahme, Elham; Gyorkos, Theresa W

    2013-05-01

    Evidence from randomized controlled trials has shown that delayed cord-clamping is beneficial to infant iron status. The role of maternal anaemia in this relationship, however, has not been established. To determine the effect of maternal anaemia at delivery on the association between timing of umbilical cord-clamping and infant anaemia at 4 and 8 months of age. A cohort of pregnant women admitted to the labour room of Hospital Iquitos (Iquitos, Peru) and their newborns were recruited into the study during two time periods (18 May to 3 June and 6-20 July 2009). Between the two recruitment periods, the hospital's policy changed from early to delayed umbilical cord-clamping. Maternal haemoglobin levels were measured before delivery, and the time between delivery and cord-clamping was recorded at delivery for the entire cohort. Mother-infant pairs were followed-up at 4 (n = 207) and 8 months (n = 184) post partum. Infant haemoglobin levels were measured at follow-up visits. Data were analysed using logistic regression models. The prevalence of maternal anaemia (Hb <11.0 g/dl) at delivery was 22%. Infant haemoglobin levels at 4 and 8 months of age were 10.4 g/dl and 10.3 g/dl, respectively. Infant haemoglobin levels did not differ significantly between infants born to anaemic mothers and those born to non-anaemic mothers at either 4 or 8 months of age. However, the association between the timing of cord-clamping and infant anaemia was modified by the mother's anaemia status. Significant benefits of delayed cord-clamping in preventing anaemia were found in infants born to anaemic mothers at both 4 months (aOR = 0.59, 95% CI 0.36-0.99) and 8 months (aOR = 0.38, 95% CI 0.19-0.76) of age. The study contributes additional evidence in support of delayed cord-clamping. This intervention is likely to have most public health impact in areas with a high prevalence of anaemia during pregnancy.

  7. Biomechanical evaluation of a new fixation device for the thoracic spine.

    PubMed

    Hongo, Michio; Ilharreborde, Brice; Gay, Ralph E; Zhao, Chunfeng; Zhao, Kristin D; Berglund, Lawrence J; Zobitz, Mark; An, Kai-Nan

    2009-08-01

    The technology used in surgery for spinal deformity has progressed rapidly in recent years. Commonly used fixation techniques may include monofilament wires, sublaminar wires and cables, and pedicle screws. Unfortunately, neurological complications can occur with all of these, compromising the patients' health and quality of life. Recently, an alternative fixation technique using a metal clamp and polyester belt was developed to replace hooks and sublaminar wiring in scoliosis surgery. The goal of this study was to compare the pull-out strength of this new construct with sublaminar wiring, laminar hooks and pedicle screws. Forty thoracic vertebrae from five fresh frozen human thoracic spines (T5-12) were divided into five groups (8 per group), such that BMD values, pedicle diameter, and vertebral levels were equally distributed. They were then potted in polymethylmethacrylate and anchored with metal screws and polyethylene bands. One of five fixation methods was applied to the right side of the vertebra in each group: Pedicle screw, sublaminar belt with clamp, figure-8 belt with clamp, sublaminar wire, or laminar hook. Pull-out strength was then assessed using a custom jig in a servohydraulic tester. The mean failure load of the pedicle screw group was significantly larger than that of the figure-8 clamp (P = 0.001), sublaminar belt (0.0172), and sublaminar wire groups (P = 0.04) with no significant difference in pull-out strength between the latter three constructs. The most common mode of failure was the fracture of the pedicle. BMD was significantly correlated with failure load only in the figure-8 clamp and pedicle screw constructs. Only the pedicle screw had a statistically significant higher failure load than the sublaminar clamp. The sublaminar method of applying the belt and clamp device was superior to the figure-8 method. The sublaminar belt and clamp construct compared favorably to the traditional methods of sublaminar wires and laminar hooks, and should be considered as an alternative fixation device in the thoracic spine.

  8. Biomechanical evaluation of a new fixation device for the thoracic spine

    PubMed Central

    Hongo, Michio; Ilharreborde, Brice; Zhao, Chunfeng; Zhao, Kristin D.; Berglund, Lawrence J.; Zobitz, Mark; An, Kai-Nan

    2009-01-01

    The technology used in surgery for spinal deformity has progressed rapidly in recent years. Commonly used fixation techniques may include monofilament wires, sublaminar wires and cables, and pedicle screws. Unfortunately, neurological complications can occur with all of these, compromising the patients’ health and quality of life. Recently, an alternative fixation technique using a metal clamp and polyester belt was developed to replace hooks and sublaminar wiring in scoliosis surgery. The goal of this study was to compare the pull-out strength of this new construct with sublaminar wiring, laminar hooks and pedicle screws. Forty thoracic vertebrae from five fresh frozen human thoracic spines (T5–12) were divided into five groups (8 per group), such that BMD values, pedicle diameter, and vertebral levels were equally distributed. They were then potted in polymethylmethacrylate and anchored with metal screws and polyethylene bands. One of five fixation methods was applied to the right side of the vertebra in each group: Pedicle screw, sublaminar belt with clamp, figure-8 belt with clamp, sublaminar wire, or laminar hook. Pull-out strength was then assessed using a custom jig in a servohydraulic tester. The mean failure load of the pedicle screw group was significantly larger than that of the figure-8 clamp (P = 0.001), sublaminar belt (0.0172), and sublaminar wire groups (P = 0.04) with no significant difference in pull-out strength between the latter three constructs. The most common mode of failure was the fracture of the pedicle. BMD was significantly correlated with failure load only in the figure-8 clamp and pedicle screw constructs. Only the pedicle screw had a statistically significant higher failure load than the sublaminar clamp. The sublaminar method of applying the belt and clamp device was superior to the figure-8 method. The sublaminar belt and clamp construct compared favorably to the traditional methods of sublaminar wires and laminar hooks, and should be considered as an alternative fixation device in the thoracic spine. PMID:19404687

  9. Fiber optic to integrated optical chip coupler

    NASA Technical Reports Server (NTRS)

    Pikulski, Joseph I. (Inventor); Ramer, O. Glenn (Inventor)

    1987-01-01

    Optical fibers are clamped by a block onto a substrate. Thereupon, metal is plated over the fibers to hold them in place upon the substrate. The clamp block is removed and the opening, resulting from the clamp block's presence, is then plated in. The built-up metallic body is a coupling which holds the fibers in position so that the ends can be polished for coupling to an integrated optical chip upon a coupling fixture.

  10. The Anion Paradox in Sodium Taste Reception: Resolution by Voltage-Clamp Studies

    NASA Astrophysics Data System (ADS)

    Ye, Qing; Heck, Gerard L.; Desimone, John A.

    1991-11-01

    Sodium salts are potent taste stimuli, but their effectiveness is markedly dependent on the anion, with chloride yielding the greatest response. The cellular mechanisms that mediate this phenomenon are not known. This "anion paradox" has been resolved by considering the field potential that is generated by restricted electrodiffusion of the anion through paracellular shunts between taste-bud cells. Neural responses to sodium chloride, sodium acetate, and sodium gluconate were studied while the field potential was voltage-clamped. Clamping at electronegative values eliminated the anion effect, whereas clamping at electropositive potentials exaggerated it. Thus, field potentials across the lingual epithelium modulate taste reception, indicating that the functional unit of taste reception includes the taste cell and its paracellular microenvironment.

  11. Hemiarch Reconstruction Vs Clamped Aortic Anastomosis for Concomitant Ascending Aortic Aneurysm.

    PubMed

    Sultan, Ibrahim; Bianco, Valentino; Yajzi, Ibrahim; Kilic, Arman; Dufendach, Keith; Cardounel, Arturo; Althouse, Andrew D; Masri, Ahmad; Navid, Forozan; Gleason, Thomas G

    2018-05-03

    Deep hypothermic circulatory arrest (DHCA) is often avoided in patients with concomitant ascending aortic pathology when treating other cardiac disease to avoid increased risk of morbidity and mortality. We hypothesized that the use of DHCA with retrograde cerebral perfusion (RCP) does not add incremental risk to ascending aortic replacement alone in the setting of concomitant cardiac surgery. 408 ascending aortic ± hemiarch replacements and aortic (root)/mitral/tricuspid valve(s), CABG, or MAZE procedures were performed for concomitant cardiac disease. DHCA with RCP was used for all hemiarch replacements or the ascending aorta was replaced with an aortic cross-clamp proximal to the innominate artery. Propensity-score matching was used to match similar ascending patients vs. hemiarch patients; the final propensity score matched patients on age, gender, BMI, previous heart surgery, pre-op aortic insufficiency, pre-op aortic stenosis, pre-op EF, and operative variables. Propensity-score matching yielded 116 pairs of Non-hemiarch patients vs. 116 hemiarch patients. Within the propensity-score matched cohort, there were no differences in postoperative stroke (1.7% vs. 3.4%, p = 0.41), new postoperative dialysis (6.0% vs. 5.2%, p = 0.78), postoperative renal insufficiency (27.6% vs. 19.8%, p = 0.16), 30-day mortality (2.6% vs. 3.4%, p = 0.701), or 1-year mortality (4.3% vs. 4.3%, p = 1.00) CONCLUSIONS: Hemiarch replacement using DHCA with RCP does not increase the risk of operative complications compared to a normothermic, clamped-distal aortic anastomosis, and therefore its use should not be limited when planning complex multi-procedural reconstructions during elective ascending thoracic aortic replacement with concomitant cardiac surgery. Copyright © 2018. Published by Elsevier Inc.

  12. Alteration of the fast excitatory postsynaptic current by barium in voltage-clamped amphibian sympathetic ganglion cells.

    PubMed Central

    Connor, E. A.; Parsons, R. L.

    1984-01-01

    Barium-induced alterations in fast excitatory postsynaptic currents (e.p.s.cs) have been studied in voltage-clamped bullfrog sympathetic ganglion B cells. In the presence of 2-8 mM barium, e.p.s.c. decay was prolonged and in many cells the e.p.s.c. decay phase deviated from a single exponential function. The decay phase in these cases was more accurately described as the sum of two exponential functions. The frequency of occurrence of a complex decay increased both with increasing barium concentration and with hyperpolarization. Miniature e.p.s.c. decay also was prolonged in barium-treated cells. E.p.s.c. amplitude was not markedly affected by barium (2-8 mM) in cells voltage-clamped to -50 mV whereas at -90 mV there was a progressive increase in peak size with increasing barium concentration. In control cells the e.p.s.c.-voltage relationship was linear between -20 and -100 mV; however, this relationship became progressively non-linear with membrane hyperpolarization in barium-treated cells. The e.p.s.c. reversal potential was shifted to a more negative value in the presence of barium. There was a voltage-dependent increase in charge movement during the e.p.s.c. in barium-treated cells which was not present in control cells. We conclude that the voltage-dependent alteration in e.p.s.c. decay time course, peak amplitude and charge movement in barium-treated cells is due to a direct postsynaptic action of barium on the kinetics of receptor-channel gating in postganglionic sympathetic neurones. PMID:6333261

  13. Cardiac troponin I is modified in the myocardium of bypass patients.

    PubMed

    McDonough, J L; Labugger, R; Pickett, W; Tse, M Y; MacKenzie, S; Pang, S C; Atar, D; Ropchan, G; Van Eyk, J E

    2001-01-02

    Selective proteolysis of cardiac troponin I (cTnI) is a proposed mechanism of contractile dysfunction in stunned myocardium, and the presence of cTnI degradation products in serum may reflect the functional state of the remaining viable myocardium. However, recent swine and canine studies have not demonstrated stunning-dependent cTnI degradation. To address the universality of cTnI modification, myocardial biopsy samples were obtained from coronary artery bypass patients (n=37) before and 10 minutes after removal of cross-clamp. Analysis of biopsy samples for cTnI by Western blotting revealed a spectrum of modified cTnI products in myocardium both before and after cross-clamp, including degradation products (7 products resulting from differential N- and C-terminal processing) and covalent complexes (3 products). In particular, a 22-kDa cTnI degradation product with C-terminal proteolysis was identified, which may represent an initial ischemia-dependent cTnI modification, similar to cTnI(1-193) observed in stunned rat myocardium. Although no systematic change in amount of modified cTnI was observed, subgroups of patients displayed an increase (n=10, 85+/-5% of cTnI remaining intact before cross-clamp versus 75+/-5% after) or a decrease (n=12, 67+/-5% before versus 78+/-5% after). Electron microscopy demonstrated normal ultrastructure in biopsy samples, which suggests no necrosis was present. In addition, cTnI modification products were observed in serum through a modified SDS-PAGE methodology. cTnI modification, in particular proteolysis, occurs in myocardium of bypass patients and may play a key role in stunning in some bypass patients.

  14. Kinetics and components of the flash photocurrent of isolated retinal rods of the larval salamander, Ambystoma tigrinum.

    PubMed Central

    Cobbs, W H; Pugh, E N

    1987-01-01

    1. Membrane currents initiated by intense, 20 microseconds flashes (photocurrents) were recorded from isolated salamander rods by combined extracellular suction electrodes and intracellular tight-seal electrodes either in current or voltage clamp mode. The magnitudes (mean +/- 2 S.E.M.) of the maximal photoresponses recorded by the suction and by the intracellular electrode respectively were 40 +/- 5 pA (n = 18) and 35 +/- 7 mV (n = 8) for current clamp at zero current; 43 +/- 9 pA and 66 +/- 13 (n = 11) pA for voltage clamp at the zero-current holding potential, -24 +/- 3 mV. 2. Photocurrents initiated by flashes isomerizing 0.1% or more of the outer segment's rhodopsin achieved a saturated velocity and were 95% complete in less than 50 ms. The effect of incrementing flash intensity above 0.1% isomerization can be described as a translation of the photocurrent along the time axis towards the origin. Within the interval 0-50 ms the latter two-thirds of the velocity-saturated photocurrent is well described as a single-exponential decay. The decay was much faster in voltage clamp (2.8 +/- 1.2 ms, n = 11) than in current clamp mode (17 +/- 5 ms, n = 17). 3. The initial third of the velocity-saturated photocurrent, occurring over the interval from the flash to the onset of exponential decay, followed about the same time course in current and voltage clamp. The time interval occupied by this initial 'latent' phase decreased with increasing flash intensity and attained an apparent minimum of about 7 ms in response to flashes isomerizing 10% or more of the rhodopsin at ca. 22 degrees C. 4. The hypothesis that the decay of outer segment light-sensitive membrane current is the same in current and voltage clamp was supported by an analysis of the difference between outer segment currents measured successively in the two recording modes. First, the tail of the difference current decayed exponentially with a time constant approximately equal to R x C, where R and C are independently estimated slope resistance and capacitance of the rod. Secondly, the integral of the difference current, when divided by outer segment capacitance, closely approximated the hyperpolarizing light response measured under current clamp. Thus, displacement current accounted for the difference between photocurrents measured in current and voltage clamp.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:2832596

  15. Long-Time Choledochal Clamping in Wistar Rats Causes Biliary Obstruction Progressing to Hepatic Fibrosis.

    PubMed

    Jorge, G D L; Tártaro, R R; Escanhoela, C A F; Boin, I D F S F

    2016-09-01

    Biliary complications are important causes of morbidity and mortality in patients undergoing hepatic surgery. The aim of the study was to evaluate late liver alterations after a long period of choledochal clamping in Wistar rats. Ten male Wistar rats, weighing 304 grams, anesthetized with sodium thiopental (25 mg/kg) and xylazine (10 mg/kg) intravenously, were distributed into 2 groups: the choledochal clamping group (CCG) and the operation sham group (OSG), with 5 animals each submitted to an abdominal incision. In the CCG, the choledochal was isolated, dissected, and clamped with a microvascular clamp for 40 minutes. After this occlusion time, the clamp was removed and the incision was closed. In the OSG the animals, under normal conditions, were submitted only to anesthesia and laparotomy for choledochal manipulation. In all animals, after the 31st day, a hepatic biopsy was carried out for histology and blood biochemical tests: total bilirubin, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyl transferase. The animals were euthanized under anesthesia. This research was approved by the Ethics Committee on Animal Use (CEUA, Unicamp, No. 2511-1). In the CCG, 100% of the animals showed bile duct dilatation, ductular proliferation, and portal inflammatory infiltrate; 60% showed regenerative nodule formation; and 80% had porta-porta septa and foci of necrosis, all of which were not found in the OSG. All CCG group biochemical tests had significant increases (P < .05) compared with OSG. Long-time choledochal clamping in Wistar rats caused hepatic dysfunction and biochemical and histological injuries with degrees of distortion to the hepatic architecture. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Can robots patch-clamp as well as humans? Characterization of a novel sodium channel mutation

    PubMed Central

    Estacion, M; Choi, J S; Eastman, E M; Lin, Z; Li, Y; Tyrrell, L; Yang, Y; Dib-Hajj, S D; Waxman, S G

    2010-01-01

    Ion channel missense mutations cause disorders of excitability by changing channel biophysical properties. As an increasing number of new naturally occurring mutations have been identified, and the number of other mutations produced by molecular approaches such as in situ mutagenesis has increased, the need for functional analysis by patch-clamp has become rate limiting. Here we compare a patch-clamp robot using planar-chip technology with human patch-clamp in a functional assessment of a previously undescribed Nav1.7 sodium channel mutation, S211P, which causes erythromelalgia. This robotic patch-clamp device can increase throughput (the number of cells analysed per day) by 3- to 10-fold. Both modes of analysis show that the mutation hyperpolarizes activation voltage dependence (−8 mV by manual profiling, −11 mV by robotic profiling), alters steady-state fast inactivation so that it requires an additional Boltzmann function for a second fraction of total current (∼20% manual, ∼40% robotic), and enhances slow inactivation (hyperpolarizing shift −15 mV by human, −13 mV robotic). Manual patch-clamping demonstrated slower deactivation and enhanced (∼2-fold) ramp response for the mutant channel while robotic recording did not, possibly due to increased temperature and reduced signal-to-noise ratio on the robotic platform. If robotic profiling is used to screen ion channel mutations, we recommend that each measurement or protocol be validated by initial comparison to manual recording. With this caveat, we suggest that, if results are interpreted cautiously, robotic patch-clamp can be used with supervision and subsequent confirmation from human physiologists to facilitate the initial profiling of a variety of electrophysiological parameters of ion channel mutations. PMID:20123784

  17. Fasting insulin sensitivity indices are not better than routine clinical variables at predicting insulin sensitivity among Black Africans: a clamp study in sub-Saharan Africans

    PubMed Central

    2014-01-01

    Background We aimed to evaluate the predictive utility of common fasting insulin sensitivity indices, and non-laboratory surrogates [BMI, waist circumference (WC) and waist-to-height ratio (WHtR)] in sub-Saharan Africans without diabetes. Methods We measured fasting glucose and insulin, and glucose uptake during 80/mU/m2/min euglycemic clamp in 87 Cameroonians (51 men) aged (SD) 34.6 (11.4) years. We derived insulin sensitivity indices including HOMA-IR, quantitative insulin sensitivity check index (QUICKI), fasting insulin resistance index (FIRI) and glucose-to-insulin ratio (GIR). Indices and clinical predictors were compared to clamp using correlation tests, robust linear regressions and agreement of classification by sex-specific thirds. Results The mean insulin sensitivity was M = 10.5 ± 3.2 mg/kg/min. Classification across thirds of insulin sensitivity by clamp matched with non-laboratory surrogates in 30-48% of participants, and with fasting indices in 27-51%, with kappa statistics ranging from −0.10 to 0.26. Fasting indices correlated significantly with clamp (/r/=0.23-0.30), with GIR performing less well than fasting insulin and HOMA-IR (both p < 0.02). BMI, WC and WHtR were equal or superior to fasting indices (/r/=0.38-0.43). Combinations of fasting indices and clinical predictors explained 25-27% of variation in clamp values. Conclusion Fasting insulin sensitivity indices are modest predictors of insulin sensitivity measured by euglycemic clamp, and do not perform better than clinical surrogates in this population. PMID:25106496

  18. Randomized clinical trial of stapler versus clamp-crushing transection in elective liver resection.

    PubMed

    Rahbari, N N; Elbers, H; Koch, M; Vogler, P; Striebel, F; Bruckner, T; Mehrabi, A; Schemmer, P; Büchler, M W; Weitz, J

    2014-02-01

    Various devices have been developed to facilitate liver transection and reduce blood loss in liver resections. None of these has proven superiority compared with the classical clamp-crushing technique. This randomized clinical trial compared the effectiveness and safety of stapler transection with that of clamp-crushing during open liver resection. Patients admitted for elective open liver resection between January 2010 and October 2011 were assigned randomly to stapler transection or the clamp-crushing technique. The primary endpoint was the total amount of intraoperative blood loss. Secondary endpoints included transection time, duration of operation, complication rates and resection margins. A total of 130 patients were enrolled, 65 to clamp-crushing and 65 to stapler transection. There was no difference between groups in total intraoperative blood loss: median (i.q.r.) 1050 (525-1650) versus 925 (450-1425) ml respectively (P = 0·279). The difference in total intraoperative blood loss normalized to the transection surface area was not statistically significant (P = 0·092). Blood loss during parenchymal transection was significantly lower in the stapler transection group (P = 0·002), as were the parenchymal transection time (mean(s.d.) 30(21) versus 9(7) min for clamp-crushing and stapler transection groups respectively; P < 0·001) and total duration of operation (mean(s.d.) 221(86) versus 190(85) min; P = 0·047). There were no significant differences in postoperative morbidity (P = 0·863) or mortality (P = 0·684) between groups. Stapler transection is a safe technique but does not reduce intraoperative blood loss in elective liver resection compared with the clamp-crushing technique. NCT01049607 (http://www.clinicaltrials.gov). © 2014 BJS Society Ltd. Published by John Wiley & Sons Ltd.

  19. Piezoresistive cantilever force-clamp system

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

    Park, Sung-Jin; Petzold, Bryan C.; Pruitt, Beth L.

    2011-04-15

    We present a microelectromechanical device-based tool, namely, a force-clamp system that sets or ''clamps'' the scaled force and can apply designed loading profiles (e.g., constant, sinusoidal) of a desired magnitude. The system implements a piezoresistive cantilever as a force sensor and the built-in capacitive sensor of a piezoelectric actuator as a displacement sensor, such that sample indentation depth can be directly calculated from the force and displacement signals. A programmable real-time controller operating at 100 kHz feedback calculates the driving voltage of the actuator. The system has two distinct modes: a force-clamp mode that controls the force applied to amore » sample and a displacement-clamp mode that controls the moving distance of the actuator. We demonstrate that the system has a large dynamic range (sub-nN up to tens of {mu}N force and nm up to tens of {mu}m displacement) in both air and water, and excellent dynamic response (fast response time, <2 ms and large bandwidth, 1 Hz up to 1 kHz). In addition, the system has been specifically designed to be integrated with other instruments such as a microscope with patch-clamp electronics. We demonstrate the capabilities of the system by using it to calibrate the stiffness and sensitivity of an electrostatic actuator and to measure the mechanics of a living, freely moving Caenorhabditis elegans nematode.« less

  20. An accurate online calibration system based on combined clamp-shape coil for high voltage electronic current transformers

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

    Li, Zhen-hua; Li, Hong-bin; Zhang, Zhi

    Electronic transformers are widely used in power systems because of their wide bandwidth and good transient performance. However, as an emerging technology, the failure rate of electronic transformers is higher than that of traditional transformers. As a result, the calibration period needs to be shortened. Traditional calibration methods require the power of transmission line be cut off, which results in complicated operation and power off loss. This paper proposes an online calibration system which can calibrate electronic current transformers without power off. In this work, the high accuracy standard current transformer and online operation method are the key techniques. Basedmore » on the clamp-shape iron-core coil and clamp-shape air-core coil, a combined clamp-shape coil is designed as the standard current transformer. By analyzing the output characteristics of the two coils, the combined clamp-shape coil can achieve verification of the accuracy. So the accuracy of the online calibration system can be guaranteed. Moreover, by employing the earth potential working method and using two insulating rods to connect the combined clamp-shape coil to the high voltage bus, the operation becomes simple and safe. Tests in China National Center for High Voltage Measurement and field experiments show that the proposed system has a high accuracy of up to 0.05 class.« less

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